WO2008049942A1 - Enzymatic collagen hydrolysate and method for obtaining same - Google Patents
Enzymatic collagen hydrolysate and method for obtaining same Download PDFInfo
- Publication number
- WO2008049942A1 WO2008049942A1 PCT/ES2007/000593 ES2007000593W WO2008049942A1 WO 2008049942 A1 WO2008049942 A1 WO 2008049942A1 ES 2007000593 W ES2007000593 W ES 2007000593W WO 2008049942 A1 WO2008049942 A1 WO 2008049942A1
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- WO
- WIPO (PCT)
- Prior art keywords
- collagen
- hydrolyzate
- enzymatic
- solution
- functional food
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- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 235000015099 wheat brans Nutrition 0.000 description 1
- 235000008939 whole milk Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
- A23C19/09—Other cheese preparations; Mixtures of cheese with other foodstuffs
- A23C19/0921—Addition, to cheese or curd, of minerals, including organic salts thereof, trace elements, amino acids, peptides, protein hydrolysates, nucleic acids, yeast extracts or autolysate, vitamins or derivatives of these compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1315—Non-milk proteins or fats; Seeds, pulses, cereals or soja; Fatty acids, phospholipids, mono- or diglycerides or derivatives therefrom; Egg products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/001—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste
- A23J1/002—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste from animal waste materials
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/10—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from hair, feathers, horn, skins, leather, bones, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
- A61K38/014—Hydrolysed proteins; Derivatives thereof from animals from connective tissue peptides, e.g. gelatin, collagen
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Definitions
- the present invention relates to an enzymatic collagen hydrolyzate with a molecular weight between 1,000 and 50,000 Daltons. Due to its special characteristics and conditions of production, it is especially suitable for use in the production of dairy products.
- Collagen is a molecule of protein nature, of fibrillar conformation and present in variable quantity in all types of connective tissue, be it skin, muscle, tendons, cartilage, blood vessels, ligaments or others.
- the first route consists in the direct extraction of collagen-rich tissues, such as skin, bones and cartilage by means of direct hydrolysis with proteolytic enzymes, followed by multiple stages of purification, until reaching a final product that is subjected to drying.
- collagen-rich tissues such as skin, bones and cartilage
- the collagen hydrolyzate is obtained from extraction methods in the absence of enzymes by hydrolysis in acidic or basic media starting from skin, bones and / or cartilage from which the gelatins have already been extracted in aqueous medium (products of higher value in the market, mainly used in food and pharmaceutical industry).
- the collagen hydrolysates thus obtained have irregular molecular weights, unpleasant taste and smell, dark color and appreciable levels of biogenic amines such as cadaverine or putrescine.
- the cost of obtaining collagen hydrolyzate by this second route is lower, but unfortunately, due to the poor organoleptic characteristics of the hydrolyzate obtained, the field of application thereof is quite limited.
- Another of the disadvantages of the usual ways of obtaining collagen hydrolyzate is that, in both cases, the collagen richness of the product obtained is less than 98% on dry product, which makes them generally unprofitable. Furthermore, taking into account the sources of supply used, preferably bovine and fish skins, it is very common for the hydrolyzate to be contaminated with impurities and unconventional agents or contaminants.
- collagen hydrolysates are widely used in many and varied fields, for example in the field of cosmetics, in the paper industry, in the food industry or in the pharmaceutical industry.
- the collagen hydrolyzate In the case of the pharmaceutical and food industry, it is required that the collagen hydrolyzate be as pure as possible, which, as already mentioned before, greatly increases the process of obtaining it.
- the situation is more unfavorable, due to the sensitivity of the stability of the milk to changes in salinity, pH, acidity and other parameters.
- the usual collagen hydrolysates by the process of obtaining them in which the protein molecules are hydrolyzed, releasing the acidic ends of the amino acids that integrate them, have such high levels of acidity, which are incompatible with the Pasteurization of mixtures of these with milk or milk derivatives at the concentrations to be used for the manufacture of said mixtures, said concentrations being between 2% and 15% by weight of hydrolyzate with respect to the total weight of the mixture.
- the researchers of the present invention have developed a method of obtaining a collagen hydrolyzate with high advantages over current procedures, while it is more economical, and have discovered an application of the obtained hydrolyzate that is also highly beneficial for the population.
- nutritional supplement should be understood all that additional dietary intake that is consumed orally and that contains a "food ingredient" intended to complement the diet.
- Some examples of dietary or nutritional supplements are vitamins, minerals, plant products (a single species or a mixture of several), other botanicals, amino acids and food components such as enzymes.
- nutritional supplements are distributed in the form of different presentations, such as tablets, capsules, soft capsules, liquids and powders.
- Functional foods are considered to be those foods, which are consumed as part of a normal diet and contain biologically active components, which offer health benefits and reduce the risk of disease.
- Some examples of functional foods include foods that contain certain minerals, vitamins, fatty acids or dietary fiber, foods to which biologically active substances, such as phytochemicals or other antioxidants, and probiotics, which have live cultures of beneficial microorganisms have been added.
- Examples of functional foods are some yogurts with probiotics, some marijuana enriched with ester esters and eggs rich in omega-3 fatty acids.
- quality of life means all that evaluation of the experience that subjects have of their own lives.
- quality of life means all that evaluation of the experience that subjects have of their own lives.
- the majority of authors conceive the quality of life as a complex and multifactorial construction on which some forms of objective measurement can be developed through a series of indicators, but where the experience that the subject may have has an important specific weight. of himself.
- Some of the indicators to determine or assess the quality of life of a subject are physical function and role, body pain, general health, vitality, etc.
- Fig. 1 shows the graph of a chromatogram corresponding to the distribution of the molecular weights of a collagen hydrolyzate according to the state of the art
- Fig. 2 represents a chromatogram corresponding to the distribution of the molecular weights of a collagen hydrolyzate according to the invention and obtainable by the described procedure.
- the collagen hydrolyzate object of the present invention with a very low total acidity and with a suitable molecular weight profile, comprised between 1,000 Daltons and 50,000 Daltons, is essentially characterized in that it has a total acidity, expressed in Dornic degrees, less than 25 0 D; and a conductivity in solution at 1% by weight not exceeding 500 ⁇ S / cm.
- Dornic grade is understood as sodium hydroxide at a concentration of 0.111 N consumed in a 10g titration of collagen enzyme hydrolyzate, where the end point marks the turn of phenolphthalein.
- the total acidity, expressed in Dornic degrees, is less than or equal to 2O 0 D.
- the enzymatic collagen hydrolyzate according to the invention is also characterized in that it has a molecular weight between 5,000 and 20,000 Daltons. According to another characteristic of the invention, the collagen enzyme hydrolyzate comprises 99% by weight of hydrolyzate on dry product.
- the enzymatic collagen hydrolyzate according to the invention is obtained by a combination of enzymatic hydrolysis and membrane filtration reactions that allow the achievement of a final product with a very low total acidity without modifying the molecular weight profile of the product.
- a method of obtaining a collagen hydrolyzate of low total acidity which starting from dermal tissue of animal origin, processed in its native form, or previously subjected to aqueous extraction, to different temperatures, once or repeatedly, and after acidic or basic hydrolysis, is characterized in that it comprises the steps of: a) aqueous extraction of the dermal tissue at a temperature between 4O 0 C and 14O 0 C for a time between 2 and 8 hours; b) pH adjustment between 6.0 and 8.0 by means of solutions of acids or mineral bases; c) filtration of the solution obtained in step b); d) demineralization of the filtered solution by passing it through ion exchange resins and / or dialysis equipment by membranes; e) ultrafiltration of the demineralized solution through membranes with a pore diameter of 1,000 to 50,000 Daltons; f) enzymatic hydrolysis of the ultrafiltered solution with an amount of enzyme comprised between 1,000 Ul and 4,000
- step f) the enzymatic hydrolysis of the ultrafiltered solution is carried out with an amount of enzyme between 1,500 Ul and 2,500 Ul per kilogram of dry product to be hydrolyzed and at a pH between the optimum pH of the enzyme ⁇ 0.5 pH units.
- step a) aqueous extraction of the raw material is performed at a temperature between 45 0 C and 100 0 C, particularly between 5O 0 C and 8O 0 C and preferably for a time between 4 and 6 hours.
- the process for obtaining total low acidity collagen hydrolyzate according to the invention is also characterized in that after the aqueous extraction the pH of the solution is adjusted between 7.4 and 7.6.
- the demineralization stage by dialysis systems is prolonged until an absolute conductivity of less than 1000 ⁇ S / cm is achieved.
- the process for obtaining collagen hydrolyzate is characterized in that the ultrafiltration stage e) of the enzymatically hydrolyzed collagen solution is carried out through membranes with a pore diameter of 5,000 to 20,000 Daltons.
- the concentration stage i) of the collagen enzymatic hydrolyzate solution is carried out at a temperature lower than or equal to 6O 0 C.
- Said concentration stage i) is carried out , preferably, at a pressure between 1.0 and 0.01 mm Hg.
- Another object of the present invention is a functional food comprising the enzymatic collagen hydrolyzate described above.
- Said hydrolyzate has a molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons, a total acidity expressed in Dornic degrees, lower at 25 0 D, a conductivity in solution at 1% by weight not exceeding 500 ⁇ S / cm and 99% by weight of collagen hydrolyzate on dry product.
- the functional food is characterized in that it comprises raw or heat treated milk.
- the functional food is also characterized by containing a compound selected independently from: antioxidant compounds, mineral salts, vitamins and / or coenzymes, plant extracts, animal extracts and adjuvant compounds of the collagen enzyme hydrolyzate, or a mixture of all of them.
- Another object of the present invention is a milk-based preparation, or a nutritional supplement, comprising an enzymatic collagen hydrolyzate having a molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons, comprising 99% in Collagen hydrolyzate weight on dry product, has a total acidity expressed in Dornic degrees, less than 25 0 D, and a conductivity in 1% solution not exceeding 500 ⁇ S / cm.
- the nutritional supplement according to the invention is also characterized in that it also comprises a compound independently selected from: antioxidant compounds, mineral salts, vitamins and / or coenzymes, plant extracts, animal extracts and adjuvant compounds of the collagen enzyme hydrolyzate, or a Mixing all of them
- the object of the present invention is also the use of an enzymatic collagen hydrolyzate having a molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons, with 99% by weight of collagen hydrolyzate on product dry, and with a total acidity expressed in Dornic degrees, less than 25 0 D, and a conductivity in solution at 1% by weight not exceeding 500 ⁇ S / cm, for the preparation of a dairy product, nutritional supplement or functional food for improve the functional capacity and / or the quality of life of a mammal with functional difficulties derived from alteration ions of articular cartilage.
- the present invention also aims at the use of the enzyme hydrolyzate Collagen agent for the preparation of a dairy product, nutritional supplement or functional food to improve physical function.
- milk itself must be understood in all its forms (whole, skimmed, semi-skimmed, etc.) and any milk derivative such as cheeses, margarines, creams, etc.
- Another object of the present invention is the use of the collagen enzymatic hydrolyzate for the preparation of a dairy product, nutritional supplement or functional food to mitigate the sensation of body pain.
- the present invention also aims at the use of the enzymatic collagen hydrolyzate to prepare a dairy product, nutritional supplement or functional food to improve the general health.
- Another object of the present invention is the use of the enzymatic collagen hydrolyzate to prepare a dairy product, nutritional supplement or functional food to improve vitality.
- the invention also relates to the use of the enzymatic collagen hydrolyzate for the preparation of a dairy product, nutritional supplement or functional food to improve social function, emotional role and / or mental health.
- the use according to the invention is further characterized in that the daily dose of enzymatic collagen hydrolyzate is comprised between 0.1 and 0.25 g / kg body weight.
- the use according to the invention is also characterized in that it is for a preparation in the form of an oral solution or suspension.
- the enzymatic collagen hydrolyzate according to the invention is for a preparation in the form of tablets or powders dispersible in water or milk.
- the use of the enzymatic collagen hydrolyzate according to the invention is characterized in that the mammal with functional difficulties derived from alterations of the articular cartilage is a human.
- a low acidic collagen enzyme hydrolyzate according to the invention, based on a possible method of obtaining it, also object of the invention, highlighting the advantages of both the hydrolyzate and its process of obtaining.
- dermal tissue of animal origin obtained under hygienic conditions, from animal species suitable for human consumption (cattle, pigs, birds and fish) is used.
- Said tissues can be processed in their native form, or previously subjected to aqueous extraction, at different temperatures, once or repeatedly, and subsequently acidic or basic hydrolysis. At this point, the tissues contain amounts of collagen ranging between 2% and 12%, those with a residual content exceeding 5% being especially valid.
- the aqueous fraction is subjected to selective precipitation of the non-collagenic proteins, elimination of these by filtration, demineralization, drying and milling.
- the solid fraction which in normal conditions would represent an important problem, due to its high organic matter load, becomes, according to this invention, the raw material of a process for obtaining collagen hydrolyzate of low total acidity.
- the raw material described above is subjected to a plurality of stages until obtaining an enzymatic hydrolyzate of high purity collagen, of weight Molecular controlled and with particular and special physical-chemical characteristics that make it suitable for use in the production of dairy products, especially those that are subjected to heat treatments.
- the starting raw material, both the dermal tissue in a native form, and the byproduct of gelatin production processes described above, are subjected to an aqueous extraction at a temperature between 4O 0 C and 14O 0 C and for a time between 2 and 8 hours.
- the aqueous extraction is performed between 45 0 C and 100 0 C; and more preferably between 5O 0 C and 8O 0 C.
- the time of the extraction can also vary and preferably lasts from 2 to 6 hours.
- the pH of the extracted solution is adjusted between 6.5 and 8.0, using solutions of acids or mineral bases.
- solutions of hydrochloric acid can be used, for example. In a preferred and more precise way the pH is adjusted between 7.4 and 7.6.
- the solution is filtered, for example, by the use of porous glass filters, polyvinyl acetate membranes, nylon fibers, activated carbon or rotary diatomaceous earth filters. Obviously, combinations of all these filters can be used until a transparent product is obtained.
- Said filtered solution is subjected to a subsequent demineralization stage by being passed through ion exchange resins or dialysis equipment.
- mixed resins are used, for example, anion exchange followed by cation exchange or vice versa.
- this step it is possible to demineralize the filtered solution until obtaining a new solution with an absolute conductivity of less than 1,000 ⁇ S / cm and a total content of less than 0.5% of mineral salts, or at least a content of mineral salts by weight less than 1.0%, so that the richness in collagen on dry extract is between 99% and 99.5% by weight.
- the parameters to be controlled in this demineralization stage are the amount of solution to be demineralized, which must be between 20 and 300 liters per kilogram of resin, with a range of 80 to 120 liters of solution per kilogram of resin being preferred.
- Another important parameter is the speed of the solution to be demineralized, which must be between 5 Htros / minute / Kg of resin and 100 liters / minute / Kg of resin. Preferably, the speed used is 20 to 40 liters / minute / kg of resin.
- the collagen solution previously filtered, will be subjected to successive steps through the dialysis membranes, in purge recirculation configuration, until a stationary regime is reached in which the absolute conductivity of the solution does not exceed 1,000 ⁇ S / cm.
- the solution is ultrafiltered by means of through the passage through membranes with a pore diameter of 1,000 to 40,000 Daltons.
- membranes with a pore diameter of 1,000 to 40,000 Daltons.
- the membranes used for the ultrafiltration stage have a pore size of 20,000 Daltons.
- the resulting solution is subjected to enzymatic hydrolysis with proteolytic enzymes under adequate temperature and pH conditions so that said enzymes can act.
- enzymes such as papain, ficin or
- the special conditions of enzymatic hydrolysis are also object of the present invention.
- a collagen hydrolyzate of low acidity and molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons
- controlled doses of proteolytic enzymes are used, at controlled temperature and time and constant pH.
- the amounts of enzyme will be of the order of 1,000 Ul to 4,000 Ul per kilogram of dry product to be hydrolyzed. It has been observed that enzyme concentrations between 1,500 Ul and 2,500 Ul per kilogram of dry product are preferred for obtaining collagen hydrolysates of low total acidity.
- Working temperatures, as well as preferred hydrolysis times will depend on the enzyme, or combination of enzymes, used.
- the pH will be controlled throughout the hydrolysis to avoid a drift greater than 2 units with respect to the optimum pH of the enzyme, or combination of enzymes, preferably a pH will be maintained, during the entire hydrolysis, within a range of ⁇ 0.5 units with respect to the optimum pH.
- the range of temperatures at which the stage of enzymatic hydrolysis is carried out is usually between 15 0 C and 8O 0 C, depending on the optimum working range of Ia or enzymes. Very satisfactory results are obtained in a temperature range of 4O 0 C to 7O 0 C.
- the enzymes are allowed to act on the ultrafiltered solution for 30 to 200 minutes, preferably 90 to 120 minutes and at a pH of the solution of reaction between 5 and 9, more specifically between 6.8 and 7.8, although this value will depend on the enzymes used.
- the enzymes must be deactivated, since they are generally thermolabile enzymes, it is only possible by increasing the temperature of the product at temperatures of 90 ° C between 5 and 30 minutes.
- the collagen enzymatic hydrolyzate solution obtained is subjected to a new filtration stage in order to ensure the absence of any type of particles that could destabilize the product, as well as give a total transparency to it.
- the filter to be used can be frames and plates, deep filtration or porous glass filters.
- this stage of filtration of the collagen enzyme hydrolyzate solution is followed by an ultrafiltration stage through membranes with a pore diameter of 1,000 to 40,000 Daltons.
- the objective of this ultrafiltration is to eliminate all those peptide fractions greater than 20,000 Daltons, as well as very low molecular weight fractions ( ⁇ 1,000 Daltons).
- this ultrafiltration stage consists of a molecular filtration or exclusion to select all those collagen enzyme hydrolyzate fractions, especially those of molecular weight between 5,000 Daltons and 20,000 Daltons.
- the collagen enzyme hydrolyzate solution is concentrated by heating at temperatures below or equal to 7O 0 C in order to obtain a concentrated product that can be dried, for example, by atomization.
- the concentration stage is carried out at a temperature lower than or equal to 6O 0 C and by means of the application of vacuum in order to evaporate the water at temperatures below the boiling temperature of the latter.
- pressures ranging from 10 to 0.0001 mm Hg are used, preferably from 1.0 to 0.01 mm Hg.
- Said concentration step is terminated when final concentrations of collagen enzyme hydrolyzate in solution of 15% to 45% by weight are reached. Specifically, a concentration range of 30% to 45% by weight of collagen enzyme hydrolyzate in solution is preferred.
- the collagen enzyme hydrolyzate solution with a weight percentage of 30% to 45% is dried until a dry product is obtained with a humidity less than or equal to 8% and a density between 0.3 g / ml and 0.8 g / ml, preferably between 0.4 g / ml and 0.6 g / ml.
- This final stage of drying the product is carried out by lyophilization or by atomization.
- the process as described allows to obtain in an economic way an enzymatic hydrolyzate of collagen with constant parameters from batch to batch and highly stable. Stability studies have revealed that the said hydrolyzate has a shelf life of about 4 years when stored at a controlled temperature of 25 0 C and a relative humidity of less than 60% atmosphere.
- the collagen enzymatic hydrolyzate obtainable according to the procedure described above is characterized by its high purity and its ideal organoleptic characteristics to be administered in oral preparations. Specifically, the enzymatic collagen hydrolyzate comprises 99% by weight of hydrolyzate on dry product.
- Said collagen enzyme hydrolyzate also object of the invention, has a molecular weight between 1,000 Daltons and 50,000 Daltons, the preferred molecular weight range of 5,000 to 20,000 Daltons.
- Humidity It must be less than 8%.
- Bulk density It must be between 0.3 and 0.8 g / ml, preferably 0.4 and 0.6 g / ml.
- Acidity (Dornic degrees): Less than 25 0 D.
- Relative conductivity (1% sol.): Must be less than 500 ⁇ S / cm
- EXAMPLE 1 7,500 Kg of dermal tissue of porcine origin, obtained directly, and in hygienic conditions, from cutting plant, and kept at a temperature below 5 o C, are subjected to the following stages:
- EXAMPLE 2 7,500 Kg of dermal tissue of porcine origin, obtained directly, and in hygienic conditions, from cutting plant, and kept at a temperature below 5 o C, are subjected to the following stages:
- the enzymatic collagen hydrolyzate according to the invention with a total acidity, expressed in Dornic degrees, less than 25 ° D; and a conductivity in 1% solution not exceeding 500 ⁇ S / cm can be obtained by other procedures, such as biotechnological procedures in which, the immobilization of enzymes on removable and easily recoverable supports, or by the contribution of the enzymes of interest working "in situ" within cell units for example bacteria.
- a low acidity collagen hydrolyzate is obtained, thus being compatible with the manufacture of nutritional supplements in which milk, or milk derivatives, is comprised as the main ingredient, and in which the collagen hydrolyzate is present in concentrations between 2% and 15% by weight, without exceeding a total acidity of 50 ° in the Dornic scale.
- the mixtures of collagen and milk hydrolyzate, or milk derivatives, in the doses detailed above are stable.
- Figs. 1 and 2 two chromatograms corresponding to the molecular weight distribution of two collagen hydrolysates
- the first (Rg. 1) corresponds to a collagen hydrolyzate prepared according to the classical process
- the second (Fig. 2) corresponds to the preparation according to the process conditions detailed above.
- the corresponding acidity values are also mentioned, expressed in degrees of the Dornic scale. It can be observed that acidity compatible with the use of collagen hydrolyzate in the formulation of pasteurized and fermented milk products has been achieved, while maintaining the original molecular weight profile. The reduction in acidity is 38%, while observing that the molecular weight profile is equivalent.
- Fig. 1 the chromatogram of an enzymatic collagen hydrolyzate, prepared by the method according to the state of the art, with a total acidity of 36.37 "Dornic, is represented.
- Fig. 2 corresponds to a collagen hydrolyzate according to the invention and obtained by the process according to the invention, whose total acidity is 22.56 ° Dornic.
- Examples of functional foods and nutritional supplements According to another of the objects of the present invention, examples of functional foods and nutritional supplements comprising the enzymatic hydrolyzate of low acid collagen according to the invention, obtainable according to the invention, are detailed below. procedure described above.
- These functional foods or nutritional supplements may further comprise a compound independently selected from: antioxidant compounds, mineral salts, vitamins and / or coenzymes, plant extracts, extracts of animal origin and adjuvant compounds of the collagen enzyme hydrolyzate. A mixture of all of them can also be used.
- EXAMPLE 1 Functional food: yogurt
- the taste of yogurt is pleasant as well as its appearance and texture.
- a chocolate dessert with the following composition has been prepared:
- a drinkable yogurt has been prepared with the following composition:
- Vegetable fiber 5.0 g
- Disodium phosphate 1.5 g Fatty acid diglycerides 1.0 g
- Vitamin complex (A, D, C, group B) 0.5 g
- Vitamin complex (C, D, group B) 0.5 g Aromas 0.2 g
- the total acidity of the product expressed as the amount of sodium hydroxide consumed for the neutralization of 10 grams of collagen hydrolyzate, according to the Dornic unit scale, must be less than 50. Preferably less than or equal to 40.
- the inventors have determined a new use of the enzymatic collagen hydrolyzate obtainable by the process described above.
- the enzymatic collagen hydrolyzate is used for the preparation of a nutritional supplement or functional food to improve the functional capacity and / or the quality of life of a mammal, specifically a human, with functional difficulties derived from alterations of the articular cartilage.
- the new collagen enzyme hydrolyzate of high purity and of better bioavailability (lower molecular weight, between 5,000 and 20,000 Daltons) as a functional food or as a component of functional foods or nutritional supplements, decided to carry out a study in 100 volunteers to whom 50 of them would be given this collagen enzyme hydrolyzate in a dose of 10 g daily, equivalent to a dose between 0.1 and 0.25 g per kg of body weight of collagen enzyme hydrolyzate and the other 50 a daily dose of 1.5 g of glucosamine sulfate, equivalent to a dose between 0.015 and 0.0375 g per kg of body weight.
- Glucosamine sulfate is widely used both in Europe and in the United States at these doses for the treatment of joint disorders in general and several studies validate that the administration of glucosamine is an effective means to improve pain and joint mobility. with consecutive dysfunctions to alterations of articular cartilage. With this test it was intended to assess the effect of the new enzymatic collagen hydrolyzate in those conditions where the known non-steroidal anti-inflammatory drugs (NSAIDs) are usually used, specifically in those disorders of the functional capacity or quality of life derived from alterations of the articular cartilage.
- NSAIDs non-steroidal anti-inflammatory drugs
- glucosamine sulfate is also widely used in Europe and especially in the United States as a functional food or as a member of functional foods and nutritional supplements (the USP 24 Io monograph includes nutritional supplements) at these same doses ( 1.5 g per day) and it is ensured that body functions generally improve, mainly mechanical ones (mobility, agility) and that they improve the general state and quality of life.
- the clinical trial consisted of a multicenter, double-blind, parallel and comparative study between the new enzymatic collagen hydrolyzate and glucosamine sulfate.
- the purpose was to compare the improvement in quality of life (according to the generic SF-36 questionnaire (The MOS 36-item short-form health survey (SF-36). I. Conceptual framework and item selection. Med care 1992 Jun; 30 (6): 473-483), as well as the efficacy, safety and tolerability of both treatments administered orally for 90 consecutive days.
- the study was carried out in 100 volunteers (93 of whom were evaluable) affected by joint disorders that induced pain, decreased mobility and notable alteration and loss of quality of life.
- the subjects were distributed in two groups of 50 individuals, of which 47 (enzymatic collagen hydrolyzate) and 46 (glucoamine sulfate) were evaluated, which, after one week of washing, received either 10g of collagen enzyme hydrolyzate or or 1.5g of glucosamine sulfate daily for 90 consecutive days, being therefore the duration of the study of 3 months for each subject.
- the quality of life was measured at the beginning and at the end of both treatments in a specific way by means of the generic SF-36 questionnaire validated in many studies and bibliographic reviews and having been used in more than 4000 scientific articles over the last 20 years, originally from the Health Institute, New England Medical Center, Boston (Massachusetts, USA). In Spain, this questionnaire has been validated by Alonso and cois. (11) at the IMIM (Municipal Institute of Medical Research) in Barcelona.
- the SF-36 represents for all authors who have carried out various validations of the questionnaire, an adequate, truthful and effective instrument very useful in clinical research and practice.
- quality of life With the term "quality of life" with the SF-36, it was intended to assess aspects of joint dysfunctions consecutive to alterations of articular cartilages that are not strictly clinical, but also aspects related to the patient's daily life. It was also intended to assess how the patient's life was affected by the presence of these joint dysfunctions, always from the patient's own point of view. This assessment was multidimensional and it was generally accepted to include at least eight sections or parameters that act as indicators of functional capacity and / or quality of life.
- the parameters evaluated correspond to the facets of daily life most commonly affected by the functional difficulties derived from alterations of the articular cartilage.
- the division of these parameters allows us to clearly differentiate activities and moments of the life of people in which the pain derived from an articular alteration affects, or can affect, the correct development of activities and tasks. It also distinguishes between the involvement of Ia quality of individual life, of the relationship with other people, or in the performance of a profession.
- the parameters evaluated are, and are defined, as follows.
- the physical function is understood as the ability to perform basic activities of daily life such as personal care, basic mobility and the ability to make small or moderate efforts. This parameter allows us to assess the degree to which the alteration of articular cartilage limits us individually.
- Physical role For its part, the physical role is defined as the parameter that evaluates the ability to perform the activities of our job. It allows us to identify if our lack of health affects our work performance, or limits us or prevents us from performing normally the tasks associated with our work. This parameter allows us to assess how lack of health affects physical activities related to our environment and work performance.
- Body pain This parameter gives us the measure of perceived pain and its effect, both at work and in relation to our environment, as well as in the basic individual activities of everyday life.
- Emotional role It is understood by emotional role, and thus it is defined for this study, the degree to which the emotional problems derived from pain and the affected tion to physical capacity affect work performance, as well as the satisfactory performance of other daily activities. The assessment of the emotional role gives us very valuable information about such important factors as performance or dedication at work.
- Table 2 Results of the generic SF-36 questionnaire applied to 100 volunteers affected by joint disorders that induced a significant loss of quality of life.
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Abstract
The invention relates to an enzymatic collagenn hydrolysate having a molecular mass of between 1000 and 50000 Daltons, low acidity and conductivity no greater than 500 μS/cm in a 1 % by weight solution. The invention also relates to a method for obtaining a collagenn hydrolysate with low total acidity from animal dermal tissue obtained under hygienic conditions from animal species suitable for human consumption (cattle, pigs, poultry and fish), which is particularly suitable for producing dairy products containing same. The method includes the following steps: aqueous extraction of the residual by-product; filtration, demineralisation and ultra-filtration of the filtered solution; enzymatic hydrolysis; concentration of the enzymatic collagen hydrolysate solution and drying of the enzymatic collagen hydrolysate. The invention further relates to nutritional supplements and functional foods containing the low-acidity enzymatic collagen hydrolysate.
Description
D E S C R I P C I O N D E S C R I P C I O N
"HIDROLIZADO ENZIMÁTICO DE COLÁGENO Y PROCEDIMIENTO DE OBTENCIÓN""COLLAGEN ENZYMATIC HYDROLYZED AND OBTAINING PROCEDURE"
Sector técnico de Ia invenciónTechnical sector of the invention
La presente invención se refiere a un hidrolizado enzimático de colágeno con un peso molecular comprendido entre 1.000 y 50.000 Daltons. Por sus especiales características y condiciones de obtención es especialmente apto para su utili- zación en Ia elaboración de productos lácteos.The present invention relates to an enzymatic collagen hydrolyzate with a molecular weight between 1,000 and 50,000 Daltons. Due to its special characteristics and conditions of production, it is especially suitable for use in the production of dairy products.
También tiene por objeto un procedimiento de obtención de un hidrolizado de colágeno a partir de tejido dérmico de origen animal, obtenido en condiciones higiénicas, de especies animales aptas para el consumo humano (bovino, porcino, aves y pescado), especialmente apto para Ia elaboración de productos lácteos que Io contengan. Dichos tejidos pueden ser procesados en su forma nativa, o proceder, como subproductos, de procesos de elaboración de gelatina.It also aims at a procedure to obtain a collagen hydrolyzate from dermal tissue of animal origin, obtained under hygienic conditions, from animal species suitable for human consumption (cattle, pigs, birds and fish), especially suitable for processing of dairy products that contain it. Such tissues can be processed in their native form, or proceed, as by-products, from gelatin production processes.
Antecedentes de Ia invenciónBackground of the invention
El colágeno es una molécula de naturaleza proteica, de conformación fibrilar y presente en cantidad variable en todos los tipos de tejido conjuntivo, ya sea piel, músculo, tendones, cartílagos, vasos sanguíneos, ligamentos u otros. El colágeno está compuesto de aminoácidos unidos mediante el enlace peptídico (HN-C=O) formando secuencias de los mismos que definen péptidos de distinto peso molecular y con distintas funciones estructurales en el organismo. En estas secuencias se encuentran todos los aminoácidos esenciales, excepto el triptófano, siendo especialmente abundantes Ia glicina, Ia prolina y Ia hidroxiprolina.Collagen is a molecule of protein nature, of fibrillar conformation and present in variable quantity in all types of connective tissue, be it skin, muscle, tendons, cartilage, blood vessels, ligaments or others. Collagen is composed of amino acids linked by the peptide bond (HN-C = O) forming sequences thereof that define peptides of different molecular weight and with different structural functions in the body. In these sequences all essential amino acids are found, except tryptophan, with glycine, proline and hydroxyproline being especially abundant.
Durante mucho tiempo el modo habitual de obtención de colágeno se ha llevado a cabo por dos vías. La primera vía consiste en Ia extracción directa de tejidos ricos en colágeno, tal como piel, huesos y cartílagos mediante Ia hidrólisis dire- cta con enzimas proteolíticos, seguida de múltiples etapas de purificación, hasta llegar a un producto final que se somete a secado. Mediante esta primera vía se obtienen productos de color claro y con olor y sabor suaves. Sin embargo, el coste de obtención del hidrolizado de colágeno obtenible por esta vía resulta muy elevado, Io que limita mucho sus aplicaciones.
Según otra vía alternativa, el hidrolizado de colágeno se obtiene a partir de métodos de extracción en ausencia de enzimas mediante hidrólisis en medios ácido o básico partiendo de piel, huesos y/o cartílagos de los que previamente ya se han extraído en medio acuoso las gelatinas (productos de valor más elevado en el mer- cado, empleados en alimentación e industria farmacéutica principalmente). Los hidrolizados de colágeno así obtenidos presentan pesos moleculares irregulares, sabor y olor desagradables, color oscuro y niveles apreciables de aminas biógenas como cadaverina o putrescina. El coste de obtención de hidrolizado de colágeno por esta segunda vía es más bajo, pero lamentablemente, debido a las deficientes características organolépticas del hidrolizado obtenido, el campo de aplicación del mismo es bastante limitado.For a long time the usual way of obtaining collagen has been carried out in two ways. The first route consists in the direct extraction of collagen-rich tissues, such as skin, bones and cartilage by means of direct hydrolysis with proteolytic enzymes, followed by multiple stages of purification, until reaching a final product that is subjected to drying. Through this first route, light colored products with a mild smell and taste are obtained. However, the cost of obtaining the collagen hydrolyzate obtainable by this route is very high, which greatly limits its applications. According to another alternative route, the collagen hydrolyzate is obtained from extraction methods in the absence of enzymes by hydrolysis in acidic or basic media starting from skin, bones and / or cartilage from which the gelatins have already been extracted in aqueous medium (products of higher value in the market, mainly used in food and pharmaceutical industry). The collagen hydrolysates thus obtained have irregular molecular weights, unpleasant taste and smell, dark color and appreciable levels of biogenic amines such as cadaverine or putrescine. The cost of obtaining collagen hydrolyzate by this second route is lower, but unfortunately, due to the poor organoleptic characteristics of the hydrolyzate obtained, the field of application thereof is quite limited.
Otro de los inconvenientes de las vías habituales de obtención de hidrolizado de colágeno es que, en ambos casos, Ia riqueza en colágeno del producto obtenido es inferior al 98% sobre producto seco, Io que las hace en general poco renta- bles. Además, tomando en consideración las fuentes de suministro utilizadas, preferentemente pieles de bóvidos y de pescado, resulta muy habitual que el hidrolizado quede contaminado con impurezas y agentes no convencionales o contaminantes.Another of the disadvantages of the usual ways of obtaining collagen hydrolyzate is that, in both cases, the collagen richness of the product obtained is less than 98% on dry product, which makes them generally unprofitable. Furthermore, taking into account the sources of supply used, preferably bovine and fish skins, it is very common for the hydrolyzate to be contaminated with impurities and unconventional agents or contaminants.
Sin embargo, los hidrolizados de colágeno se emplean ampliamente en muchos y variados campos, por ejemplo en el campo de Ia cosmética, en Ia industria del papel, en Ia industria alimentaria o en Ia industria farmacéutica. En el caso de Ia industria farmacéutica y de Ia alimentaria se requiere que el hidrolizado de colágeno sea Io más puro posible, Io que, como ya se ha comentado antes, encarece notablemente el proceso de obtención. En el caso particular de Ia utilización de hidrolizado de colágeno en Ia industria láctea, Ia situación es más desfavorable, debido a Ia sensibilidad de Ia estabilidad de Ia leche a cambios de salinidad, pH, acidez y otros parámetros.However, collagen hydrolysates are widely used in many and varied fields, for example in the field of cosmetics, in the paper industry, in the food industry or in the pharmaceutical industry. In the case of the pharmaceutical and food industry, it is required that the collagen hydrolyzate be as pure as possible, which, as already mentioned before, greatly increases the process of obtaining it. In the particular case of the use of collagen hydrolyzate in the dairy industry, the situation is more unfavorable, due to the sensitivity of the stability of the milk to changes in salinity, pH, acidity and other parameters.
Los investigadores han desarrollado con anterioridad un procedimiento según el cual, a partir de subproductos de procesos de obtención de gelatina, se obtiene un hidrolizado enzimático de colágeno muy puro y aplicable tanto en Ia fabri- cación de alimentos funcionales como en suplementos nutricionales, permitiendo una mejora de Ia capacidad funcional y/o Ia calidad de vida de un mamífero con dificultades funcionales derivadas de alteraciones de los cartílagos articulares. Sin embargo, cuando dichos hidrolizados enzimáticos de colágeno deben emplearse en Ia fabricación de productos lácteos (leche, yogures, quesos, etc.), debido a Ia aci-
dez y/o salinidad de los mismos, aparecen problemas para obtener productos con Ia consistencia adecuada, por ejemplo problemas de precipitación en medio ácido de las proteínas de Ia leche, o problemas de incompatibilidad con los otros ingredientes del suplemento nutricional o alimento funcional. Hasta Ia fecha, si se quería obtener un hidrolizado enzimático de colágeno con baja acidez, los mismos quedaban supeditados a unos pesos moleculares muy elevados, que no los hacían adecuados para Ia preparación de estos productos lácteos, principalmente debido a su gran viscosidad.Researchers have previously developed a procedure according to which, from by-products of gelatin production processes, an enzymatic hydrolyzate of very pure collagen is obtained and applicable both in the manufacture of functional foods and in nutritional supplements, allowing a improvement of the functional capacity and / or the quality of life of a mammal with functional difficulties derived from alterations of the articular cartilage. However, when said enzymatic collagen hydrolysates must be used in the manufacture of dairy products (milk, yogurts, cheeses, etc.), due to the acid Dez and / or salinity thereof, problems appear to obtain products with adequate consistency, for example problems of precipitation in acidic medium of milk proteins, or problems of incompatibility with the other ingredients of the nutritional supplement or functional food. To date, if an enzymatic hydrolyzate of collagen with low acidity was to be obtained, they were subject to very high molecular weights, which did not make them suitable for the preparation of these dairy products, mainly due to their high viscosity.
Concretamente, los hidrolizados de colágeno habituales, por el propio pro- cedimiento de obtención de los mismos en que se van hidrolizando las moléculas proteicas liberándose los extremos ácidos de los aminoácidos que las integran, presentan niveles de acidez tan altos, que resultan incompatibles con Ia pasteurización de mezclas de éstos con leche o derivados lácteos a las concentraciones que deben emplearse para Ia fabricación de dichas mezclas, estando dichas concentracio- nes comprendidas entre el 2% y el 15% en peso de hidrolizado respecto del peso total de Ia mezcla.Specifically, the usual collagen hydrolysates, by the process of obtaining them in which the protein molecules are hydrolyzed, releasing the acidic ends of the amino acids that integrate them, have such high levels of acidity, which are incompatible with the Pasteurization of mixtures of these with milk or milk derivatives at the concentrations to be used for the manufacture of said mixtures, said concentrations being between 2% and 15% by weight of hydrolyzate with respect to the total weight of the mixture.
En anteriores investigaciones, y de acuerdo con Ia patente US 6,211 ,143, ya se había conseguido demostrar con hidrolizados de gelatina de menor pureza en colágeno y con pesos moleculares mayores (hasta 100.0000 daltons) que Ia ingesta del mismo por deportistas y personas con un esfuerzo especial en sus actividades normales conseguían mantener o mejorar Ia integridad de los cartílagos articulares. De Ia patente ES 2087030 se deduce que el hidrolizado de gelatina también es útil como coadyuvante en los tratamientos destinados a compensar el déficit de calcio. También se deduce otra aplicación de Ia patente ES 2104507, donde se describen composiciones para Ia preparación de medicamentos para el tratamiento y prevención de Ia displasia de cadera en perros.In previous investigations, and in accordance with US Patent 6,211, 143, it had already been demonstrated with gelatin hydrolysates of lower purity in collagen and with higher molecular weights (up to 1000000 daltons) than the intake thereof by athletes and people with a special effort in their normal activities managed to maintain or improve the integrity of the articular cartilage. From the patent ES 2087030 it follows that the gelatin hydrolyzate is also useful as an adjunct in treatments aimed at compensating for calcium deficiency. Another application of the patent ES 2104507 is also deduced, where compositions for the preparation of medicaments for the treatment and prevention of hip dysplasia in dogs are described.
Varios estudios realizados recientemente han demostrado que el hidrolizado enzimático de colágeno es bien absorbido cuando se administra por vía oral y se distribuye con gran afinidad preferentemente en los tejidos de los cartílagos articulares (Oesser S, et al. "Oral administration of 14C Labelled Gelatin hydrolysate leads to an accumulation of radioactivity in cartilage of mice (C57/BL)". J Nυtr 1999; 129: 1891-1895.). Este espectro de distribución provoca un incremento del volumen de Ia masa cartilaginosa, Ia cual puede desarrollar su acción con Ia consecuente dismi-
nución del dolor y de Ia inflamación y con Ia mejoría de Ia motilidad de las citadas articulaciones. En este aspecto se remite el lector a las siguientes publicaciones: Adam M. "Therapie der osteoarthrosis. Welche Wirkung haben gelatinepráparate" Therapiewoche 1991 , 41: 2458-2461; Arquer A, et al. "Physical exercise in the eld- erly people". Selección 1996; 5 (3): 121 -128; Ribas J, et al. "Effects of gelatine hy- drolysatesin the prevention of sportsmen and woman injuries." Arch Med Deport 1998; XV n° 66: 277-282; y Trentham et al. "Effects of oral administration of type Il collagen on rheumatoid arthritis." Science 1993; 261: 1727-1730.Several recent studies have shown that collagen enzyme hydrolyzate is well absorbed when administered orally and is distributed with great affinity preferentially in the tissues of articular cartilage (Oesser S, et al. "Oral administration of 14 C Labelled Gelatin hydrolysate leads to an accumulation of radioactivity in cartilage of mice (C57 / BL) ". J Nυtr 1999; 129: 1891-1895.). This distribution spectrum causes an increase in the volume of the cartilaginous mass, which can develop its action with the consequent decrease nución of pain and inflammation and with the improvement of the motility of said joints. In this regard, the reader refers to the following publications: Adam M. "Therapie der osteoarthrosis. Welche Wirkung haben gelatinepráparate" Therapiewoche 1991, 41: 2458-2461; Arquer A, et al. "Physical exercise in the elderly people". Selection 1996; 5 (3): 121-128; Ribas J, et al. "Effects of gelatine hy- drolysatesin the prevention of sportsmen and woman injuries." Arch Med Deport 1998; XV No. 66: 277-282; and Trentham et al. "Effects of oral administration of type Il collagen on rheumatoid arthritis." Science 1993; 261: 1727-1730.
Los investigadores de Ia presente invención, han desarrollado un procedi- miento de obtención de un hidrolizado de colágeno con elevadas ventajas respecto a los procedimientos actuales, a Ia vez que resulta más económico, y han descubierto una aplicación del hidrolizado obtenido que resulta también altamente beneficiosa para Ia población.The researchers of the present invention have developed a method of obtaining a collagen hydrolyzate with high advantages over current procedures, while it is more economical, and have discovered an application of the obtained hydrolyzate that is also highly beneficial for the population.
Cada vez más, Ia población se preocupa por Ia dietética y Ia nutrición tanto animal como humana. Con ello, han aparecido en el mercado múltiples preparados conocidos como suplementos nutricionales que aportan a los animales o las personas Ia cantidad necesaria de vitaminas, sales minerales o aminoácidos esenciales. También en el campo de los suplementos nutricionales se ha empleado el colágeno para obtener preparaciones que favorezcan Ia recuperación de los cartílagos en animales que sufren enfermedades vinculadas a este tipo de tejido.Increasingly, the population is concerned with dietary and animal and human nutrition. With this, multiple preparations known as nutritional supplements that provide animals or people with the necessary amount of vitamins, mineral salts or essential amino acids have appeared on the market. Also in the field of nutritional supplements, collagen has been used to obtain preparations that favor the recovery of cartilage in animals suffering from diseases linked to this type of tissue.
Como suplemento nutricional debe entenderse todo aquel aporte adicional dietético que se consume por vía oral y que contiene un "ingrediente alimenticio" destinado a complementar Ia alimentación. Algunos ejemplos de suplementos dietéticos o nutricionales son las vitaminas, los minerales, los productos vegetales (una sola especie o una mezcla de varias), otros productos botánicos, aminoácidos y componentes de los alimentos como las enzimas. Generalmente los suplementos nutricionales se distribuyen en forma de diferentes presentaciones, como comprimidos, cápsulas, cápsulas blandas, líquidos y polvos.As nutritional supplement should be understood all that additional dietary intake that is consumed orally and that contains a "food ingredient" intended to complement the diet. Some examples of dietary or nutritional supplements are vitamins, minerals, plant products (a single species or a mixture of several), other botanicals, amino acids and food components such as enzymes. Generally nutritional supplements are distributed in the form of different presentations, such as tablets, capsules, soft capsules, liquids and powders.
También en el campo de Ia nutrición se están dando a conocer últimamente los denominados alimentos funcionales. Los alimentos funcionales, se considera que son aquellos alimentos, que se consumen como parte de una dieta normal y contienen componentes biológicamente activos, que ofrecen beneficios para Ia salud y reducen el riesgo de sufrir enfermedades. Entre algunos ejemplos de alimentos funcionales, destacan los alimentos que contienen determinados minerales,
vitaminas, ácidos grasos o fibra alimenticia, los alimentos a los que se han añadido sustancias biológicamente activas, como los fitoquímicos u otros antioxidantes, y los probióticos, que tienen cultivos vivos de microorganismos beneficiosos. Son ejemplos de alimentos funcionales algunos yogures con probióticos, algunas mar- garinas enriquecidas con esteres de esteróles y los huevos ricos en ácidos grasos del tipo omega-3.Also in the field of nutrition, so-called functional foods are being disclosed lately. Functional foods are considered to be those foods, which are consumed as part of a normal diet and contain biologically active components, which offer health benefits and reduce the risk of disease. Some examples of functional foods include foods that contain certain minerals, vitamins, fatty acids or dietary fiber, foods to which biologically active substances, such as phytochemicals or other antioxidants, and probiotics, which have live cultures of beneficial microorganisms have been added. Examples of functional foods are some yogurts with probiotics, some marijuana enriched with ester esters and eggs rich in omega-3 fatty acids.
Tanto los alimentos funcionales como los suplementos nutricionales se asocian científicamente a Ia mejora de Ia calidad de vida de las personas o animales que los ingieren. También se han asociado a Ia sensación de bienestar general que provocan, tomados solos o como complemento a determinados tratamientos terapéuticos.Both functional foods and nutritional supplements are scientifically associated with the improvement of the quality of life of the people or animals that ingest them. They have also been associated with the feeling of general well-being that they cause, taken alone or as a complement to certain therapeutic treatments.
Aunque resulta un término difícil de definir, por calidad de vida se entiende toda aquella evaluación de Ia experiencia que de su propia vida tienen los sujetos. Así Ia mayoría de autores conciben Ia calidad de vida como una construcción com- pleja y multifactorial sobre Ia que pueden desarrollarse algunas formas de medida objetiva a través de una serie de indicadores, pero donde tiene un importante peso específico Ia vivencia que el sujeto pueda tener de sí mismo. Algunos de los indicadores para determinar o valorar Ia calida de vida de un sujeto son Ia función y rol físicos, el dolor corporal, Ia salud en general, Ia vitalidad, etc.Although it is a difficult term to define, quality of life means all that evaluation of the experience that subjects have of their own lives. Thus the majority of authors conceive the quality of life as a complex and multifactorial construction on which some forms of objective measurement can be developed through a series of indicators, but where the experience that the subject may have has an important specific weight. of himself. Some of the indicators to determine or assess the quality of life of a subject are physical function and role, body pain, general health, vitality, etc.
Breve descripción de los dibujosBrief description of the drawings
A modo de ejemplo y para ilustrar mejor el objeto de Ia presente invención, se adjuntan dos figuras en las que:By way of example and to better illustrate the object of the present invention, two figures are attached in which:
La Fig. 1 muestra el gráfico de un cromatograma correspondiente a Ia distri- bución de los pesos moleculares de un hidrolizado de colágeno según el estado de Ia técnica; yFig. 1 shows the graph of a chromatogram corresponding to the distribution of the molecular weights of a collagen hydrolyzate according to the state of the art; Y
La Fig. 2 representa un cromatograma correspondiente a Ia distribución de los pesos moleculares de un hidrolizado de colágeno según Ia invención y obtenible por el procedimiento descrito.Fig. 2 represents a chromatogram corresponding to the distribution of the molecular weights of a collagen hydrolyzate according to the invention and obtainable by the described procedure.
Explicación de Ia invenciónExplanation of the invention
El hidrolizado de colágeno objeto de Ia presente invención, con una acidez total muy baja y con un el perfil de pesos moleculares adecuado, comprendidos entre los 1.000 Daltons y los 50.000 Daltons, en su esencia se caracteriza porque
tiene una acidez total, expresada en grados Dornic, inferior a 250D; y una conductividad en solución al 1% en peso no superior a 500 μS/cm.The collagen hydrolyzate object of the present invention, with a very low total acidity and with a suitable molecular weight profile, comprised between 1,000 Daltons and 50,000 Daltons, is essentially characterized in that it has a total acidity, expressed in Dornic degrees, less than 25 0 D; and a conductivity in solution at 1% by weight not exceeding 500 μS / cm.
Se entiende por grado Dornic el hidróxido sódico a una concentración de 0,111 N consumido en una valoración de 10g de hidrolizado enzimático de coláge- no, donde el punto final Io marca el viraje de Ia fenolftaleína.Dornic grade is understood as sodium hydroxide at a concentration of 0.111 N consumed in a 10g titration of collagen enzyme hydrolyzate, where the end point marks the turn of phenolphthalein.
Según otra característica de Ia invención, Ia acidez total, expresada en grados Dornic, es inferior o igual a 2O0D.According to another characteristic of the invention, the total acidity, expressed in Dornic degrees, is less than or equal to 2O 0 D.
El hidrolizado enzimático de colágeno según Ia invención, se caracterizad también porque tiene un peso molecular comprendido entre 5.000 y 20.000 Daltons. Según otra característica de Ia invención, el hidrolizado enzimático de colágeno comprende un 99% en peso de hidrolizado sobre producto seco.The enzymatic collagen hydrolyzate according to the invention is also characterized in that it has a molecular weight between 5,000 and 20,000 Daltons. According to another characteristic of the invention, the collagen enzyme hydrolyzate comprises 99% by weight of hydrolyzate on dry product.
El hidrolizado enzimático de colágeno según Ia invención, se obtiene por una combinación de reacciones de hidrólisis enzimática y filtración con membranas que permiten Ia consecución de un producto final con una acidez total muy baja sin modificar el perfil de pesos moleculares del producto.The enzymatic collagen hydrolyzate according to the invention is obtained by a combination of enzymatic hydrolysis and membrane filtration reactions that allow the achievement of a final product with a very low total acidity without modifying the molecular weight profile of the product.
Así, es también objeto de Ia presente invención, un procedimiento de obtención de un hidrolizado de colágeno de baja acidez total según Ia invención, que partiendo de tejido dérmico de origen animal, procesado en su forma nativa, o previamente sometido a extracción acuosa, a diferentes temperaturas, una o repetidas veces, y a posterior hidrólisis acida o básica, se caracteriza porque comprende las etapas de: a) extracción acuosa del tejido dérmico de partida a una temperatura comprendida entre 4O0C y 14O0C durante un tiempo comprendido entre 2 y 8 horas; b) ajuste del pH entre 6,0 y 8,0 mediante soluciones de ácidos o bases mi- nerales; c) filtración de Ia solución obtenida en Ia etapa b); d) desmineralización de Ia solución filtrada por paso de Ia misma a través de resinas de intercambio iónico y/o equipos de diálisis por membranas; e) ultrafiltración de Ia solución desmineralizada a través de membranas con un diámetro de poro de 1.000 a 50.000 Daltons; f) hidrólisis enzimática de Ia solución ultrafiltrada con una cantidad de enzima comprendida entre 1.000 Ul y 4.000 Ul por kilogramo de producto seco a hidro- lizar y a un pH comprendido entre el pH óptimo de Ia enzima ± 0,5 unidades de pH ; g) filtración de Ia solución de colágeno hidrolizado enzimáticamente;
h) ultrafiltración a través de membranas con un diámetro de poro de 10.000 a 40.000 Daltons; i) concentración de Ia solución de hidrolizado enzimático de colágeno mediante calefacción a temperaturas inferiores o iguales a 7O0C a una presión com- prendida entre 10 y 0,0001 mm de Hg hasta una concentración final de hidrolizado enzimático de colágeno del 15% al 45% en peso; j) secado del hidrolizado enzimático de colágeno hasta obtener un producto seco con una humedad inferior o igual al 8% y una densidad entre 0,3 y 0,8 g/ml.Thus, it is also an object of the present invention, a method of obtaining a collagen hydrolyzate of low total acidity according to the invention, which starting from dermal tissue of animal origin, processed in its native form, or previously subjected to aqueous extraction, to different temperatures, once or repeatedly, and after acidic or basic hydrolysis, is characterized in that it comprises the steps of: a) aqueous extraction of the dermal tissue at a temperature between 4O 0 C and 14O 0 C for a time between 2 and 8 hours; b) pH adjustment between 6.0 and 8.0 by means of solutions of acids or mineral bases; c) filtration of the solution obtained in step b); d) demineralization of the filtered solution by passing it through ion exchange resins and / or dialysis equipment by membranes; e) ultrafiltration of the demineralized solution through membranes with a pore diameter of 1,000 to 50,000 Daltons; f) enzymatic hydrolysis of the ultrafiltered solution with an amount of enzyme comprised between 1,000 Ul and 4,000 Ul per kilogram of dry product to be hydrogenated and at a pH between the optimum pH of the enzyme ± 0.5 pH units; g) filtration of the enzymatically hydrolyzed collagen solution; h) ultrafiltration through membranes with a pore diameter of 10,000 to 40,000 Daltons; i) concentration of the collagen enzyme hydrolyzate solution by heating at temperatures below or equal to 7O 0 C at a pressure between 10 and 0.0001 mm Hg to a final concentration of 15% collagen enzyme hydrolyzate at 45% by weight; j) drying of the enzymatic collagen hydrolyzate to obtain a dry product with a humidity less than or equal to 8% and a density between 0.3 and 0.8 g / ml.
El procedimiento según Ia invención se caracteriza también porque en Ia etapa f) se realiza Ia hidrólisis enzimática de Ia solución ultrafiltrada con una cantidad de enzima comprendida entre 1.500 Ul y 2.500 Ul por kilogramo de producto seco a hidrolizar y a un pH comprendido entre el pH óptimo de Ia enzima ± 0,5 unidades de pH.The process according to the invention is also characterized in that in step f) the enzymatic hydrolysis of the ultrafiltered solution is carried out with an amount of enzyme between 1,500 Ul and 2,500 Ul per kilogram of dry product to be hydrolyzed and at a pH between the optimum pH of the enzyme ± 0.5 pH units.
Según otra característica del procedimiento de obtención de hidrolizado en- zimático de colágeno de baja acidez total, Ia etapa a) de extracción acuosa de Ia materia prima se realiza a una temperatura comprendida entre 450C y 1000C, concretamente entre 5O0C y 8O0C y preferentemente durante un tiempo comprendido entre 4 y 6 horas.According to another characteristic of the procedure for hydrolysed collagen in- zimático low total acidity, step a) aqueous extraction of the raw material is performed at a temperature between 45 0 C and 100 0 C, particularly between 5O 0 C and 8O 0 C and preferably for a time between 4 and 6 hours.
El procedimiento de obtención de hidrolizado de colágeno de baja acidez to- tal según Ia invención, se caracteriza también porque después de Ia extracción acuosa se ajusta el pH de Ia solución entre 7,4 y 7,6.The process for obtaining total low acidity collagen hydrolyzate according to the invention is also characterized in that after the aqueous extraction the pH of the solution is adjusted between 7.4 and 7.6.
En el caso particular del hidrolizado de colágeno objeto de Ia presente invención, Ia etapa de desmineralización mediante sistemas de diálisis se prolonga hasta conseguir una conductividad absoluta inferior a 1000 μS/cm. El procedimiento de obtención de hidrolizado de colágeno está caracterizado porque Ia etapa de ultrafiltración e) de Ia solución de colágeno hidrolizado enzimáti- camente se lleva a cabo a través de membranas con un diámetro de poro de 5.000 a 20.000 Daltons.In the particular case of the collagen hydrolyzate object of the present invention, the demineralization stage by dialysis systems is prolonged until an absolute conductivity of less than 1000 μS / cm is achieved. The process for obtaining collagen hydrolyzate is characterized in that the ultrafiltration stage e) of the enzymatically hydrolyzed collagen solution is carried out through membranes with a pore diameter of 5,000 to 20,000 Daltons.
Según otra característica del procedimiento de obtención de hidrolizado de colágeno según Ia invención, Ia etapa de concentración i) de Ia solución de hidrolizado enzimático de colágeno se realiza a una temperatura inferior o igual a 6O0C. Dicha etapa de concentración i) se realiza, de un modo preferido, a una presión comprendida entre 1,0 y 0,01 mm de Hg.
Otro objeto de Ia presente invención es un alimento funcional que comprende el hidrolizado enzimático de colágeno anteriormente descrito Dicho hidrolizado tiene un peso molecular comprendido entre 1.000 y 50.000 Daltons, preferentemente entre 5.000 y 20.000 Daltons, una acidez total expresada en grados Dornic, infe- rior a 250D, una conductividad en solución al 1% en peso no superior a 500 μS/cm y un 99% en peso de hidrolizado de colágeno sobre producto seco.According to another characteristic of the process for obtaining collagen hydrolyzate according to the invention, the concentration stage i) of the collagen enzymatic hydrolyzate solution is carried out at a temperature lower than or equal to 6O 0 C. Said concentration stage i) is carried out , preferably, at a pressure between 1.0 and 0.01 mm Hg. Another object of the present invention is a functional food comprising the enzymatic collagen hydrolyzate described above. Said hydrolyzate has a molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons, a total acidity expressed in Dornic degrees, lower at 25 0 D, a conductivity in solution at 1% by weight not exceeding 500 μS / cm and 99% by weight of collagen hydrolyzate on dry product.
El alimento funcional, según Ia invención, se caracteriza porque comprende leche cruda o tratada térmicamente.The functional food, according to the invention, is characterized in that it comprises raw or heat treated milk.
Además, el alimento funcional, según Ia invención, también se caracteriza por contener un compuesto seleccionado independientemente de entre: compuestos antioxidantes, sales minerales, vitaminas y/o coenzimas, extractos vegetales, extractos de origen animal y compuestos coadyuvantes del hidrolizado enzimático de colágeno, o una mezcla de todos ellos.In addition, the functional food, according to the invention, is also characterized by containing a compound selected independently from: antioxidant compounds, mineral salts, vitamins and / or coenzymes, plant extracts, animal extracts and adjuvant compounds of the collagen enzyme hydrolyzate, or a mixture of all of them.
Otro objeto de Ia presente invención es un preparado a base de leche, o un suplemento nutricional, que comprende un hidrolizado enzimático de colágeno que tiene un peso molecular comprendido entre 1.000 y 50.000 Daltons, preferentemente entre 5.000 y 20.000 Daltons, comprende un 99% en peso de hidrolizado de colágeno sobre producto seco, tiene una acidez total expresada en grados Dornic, inferior a 250D, y una conductividad en solución al 1% no superior a 500 μS/cm. El suplemento nutricional según Ia invención, se caracteriza también porque además comprende un compuesto seleccionado independientemente de entre: compuestos antioxidantes, sales minerales, vitaminas y/o coenzimas, extractos vegetales, extractos de origen animal y compuestos coadyuvantes del hidrolizado enzimático de colágeno, o una mezcla de todos ellos Es también objeto de Ia presente invención el uso de un hidrolizado enzimático de colágeno que tiene un peso molecular comprendido entre 1.000 y 50.000 Daltons, preferentemente entre 5.000 y 20.000 Daltons, con un 99% en peso de hidrolizado de colágeno sobre producto seco, y con una acidez total expresada en grados Dornic, inferior a 250D, y una conductividad en solución al 1 % en peso no superior a 500 μS/cm, para Ia preparación de un producto lácteo, suplemento nutricional o alimento funcional para mejorar Ia capacidad funcional y/o Ia calidad de vida de un mamífero con dificultades funcionales derivadas de alteraciones de los cartílagos articulares.Another object of the present invention is a milk-based preparation, or a nutritional supplement, comprising an enzymatic collagen hydrolyzate having a molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons, comprising 99% in Collagen hydrolyzate weight on dry product, has a total acidity expressed in Dornic degrees, less than 25 0 D, and a conductivity in 1% solution not exceeding 500 μS / cm. The nutritional supplement according to the invention is also characterized in that it also comprises a compound independently selected from: antioxidant compounds, mineral salts, vitamins and / or coenzymes, plant extracts, animal extracts and adjuvant compounds of the collagen enzyme hydrolyzate, or a Mixing all of them The object of the present invention is also the use of an enzymatic collagen hydrolyzate having a molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons, with 99% by weight of collagen hydrolyzate on product dry, and with a total acidity expressed in Dornic degrees, less than 25 0 D, and a conductivity in solution at 1% by weight not exceeding 500 μS / cm, for the preparation of a dairy product, nutritional supplement or functional food for improve the functional capacity and / or the quality of life of a mammal with functional difficulties derived from alteration ions of articular cartilage.
La presente invención también tiene por objeto el uso del hidrolizado enzi-
mático de colágeno para Ia preparación de un producto lácteo, suplemento nutricio- nal o alimento funcional para mejorar Ia función física.The present invention also aims at the use of the enzyme hydrolyzate Collagen agent for the preparation of a dairy product, nutritional supplement or functional food to improve physical function.
Por producto lácteo debe entenderse Ia propia leche en todas sus formas (entera, desnatada, semidesnatada, etc.) y cualquier derivado de Ia leche tal como quesos, margarinas, natas, etc.By dairy product, milk itself must be understood in all its forms (whole, skimmed, semi-skimmed, etc.) and any milk derivative such as cheeses, margarines, creams, etc.
Otro objeto de Ia presente invención es el uso del hidrolizado enzimático de colágeno para Ia preparación de un producto lácteo, suplemento nutricional o alimento funcional para atenuar Ia sensación de dolor corporal.Another object of the present invention is the use of the collagen enzymatic hydrolyzate for the preparation of a dairy product, nutritional supplement or functional food to mitigate the sensation of body pain.
La presente invención también tiene por objeto el uso del hidrolizado enzi- mático de colágeno para preparar un producto lácteo, suplemento nutricional o alimento funcional para mejorar Ia salud general.The present invention also aims at the use of the enzymatic collagen hydrolyzate to prepare a dairy product, nutritional supplement or functional food to improve the general health.
Otro objeto de Ia presente invención es el uso del hidrolizado enzimático de colágeno para preparar un producto lácteo, suplemento nutricional o alimento funcional para mejorar Ia vitalidad. La invención también se refiere al uso del hidrolizado enzimático de colágeno para Ia preparación de un producto lácteo, suplemento nutricional o alimento funcional para mejorar Ia función social, el rol emocional y/o Ia salud mental.Another object of the present invention is the use of the enzymatic collagen hydrolyzate to prepare a dairy product, nutritional supplement or functional food to improve vitality. The invention also relates to the use of the enzymatic collagen hydrolyzate for the preparation of a dairy product, nutritional supplement or functional food to improve social function, emotional role and / or mental health.
El uso según Ia invención se caracteriza además porque Ia dosis diaria de hidrolizado enzimático de colágeno está comprendida entre 0,1 y 0,25 g/Kg de peso corporal.The use according to the invention is further characterized in that the daily dose of enzymatic collagen hydrolyzate is comprised between 0.1 and 0.25 g / kg body weight.
El uso según Ia invención se caracteriza también porque es para una preparación en forma de solución o suspensión oral.The use according to the invention is also characterized in that it is for a preparation in the form of an oral solution or suspension.
Según otra característica del uso del hidrolizado enzimático de colágeno según Ia invención, el mismo es para una preparación en forma de comprimidos o polvos dispersables en agua o leche.According to another characteristic of the use of the enzymatic collagen hydrolyzate according to the invention, it is for a preparation in the form of tablets or powders dispersible in water or milk.
El uso del hidrolizado enzimático de colágeno según Ia invención, se caracteriza porque el mamífero con dificultades funcionales derivadas de alteraciones de los cartílagos articulares es un humano.The use of the enzymatic collagen hydrolyzate according to the invention is characterized in that the mammal with functional difficulties derived from alterations of the articular cartilage is a human.
Descripción detallada de Ia invenciónDetailed description of the invention
A continuación se detallan las características esenciales de un hidrolizado enzimático de colágeno de baja acidez de acuerdo con Ia invención, en base a un posible procedimiento de obtención del mismo, también objeto de Ia invención, resaltando las ventajas tanto del hidrolizado como de su procedimiento de obtención.
En el citado procedimiento se parte de tejido dérmico de origen animal, obtenido en condiciones higiénicas, de especies animales aptas para el consumo humano (bovino, porcino, aves y pescado). Dichos tejidos pueden ser procesados en su forma nativa, o previamente sometidos a extracción acuosa, a diferentes temperaturas, una o repetidas veces, y a posterior hidrólisis acida o básica. En este punto, los tejidos contienen cantidades de colágeno que oscilan entre el 2% y 12%, resultando especialmente válidos aquellos con un contenido residual superior al 5%.The essential characteristics of a low acidic collagen enzyme hydrolyzate according to the invention are detailed below, based on a possible method of obtaining it, also object of the invention, highlighting the advantages of both the hydrolyzate and its process of obtaining. In the aforementioned procedure, dermal tissue of animal origin, obtained under hygienic conditions, from animal species suitable for human consumption (cattle, pigs, birds and fish) is used. Said tissues can be processed in their native form, or previously subjected to aqueous extraction, at different temperatures, once or repeatedly, and subsequently acidic or basic hydrolysis. At this point, the tissues contain amounts of collagen ranging between 2% and 12%, those with a residual content exceeding 5% being especially valid.
En el caso particular de tejidos sometidos a extracción previa, se tratará de subproductos de procesos de obtención de gelatinas. Los procesos de extracción de gelatina clásicos se basan, a grandes rasgos, en Ia maceración del tejido dérmico en condiciones de extrema acidez o alcalinidad durante un tiempo total de 4 a 24 horas, seguido de un lavado con agua desmineralizada, una decantación de Ia grasa liberada y una neutralización con soluciones acidas o básicas inorgánicas. Pos- teriormente se procede a Ia solubilización de Ia gelatina, mediante extracción del tejido sólido con disolventes acuosos, en nuestro caso particular, el disolvente es agua desmineralizada. En este punto se separa Ia solución de gelatina del resto sólido. La fracción acuosa se somete a precipitación selectiva de las proteínas no colagénicas, eliminación de éstas por filtración, desmineralización, secado y moltu- ración. La fracción sólida, que en condiciones normales representaría un importante problema, debido a su elevada carga de materia orgánica, se convierte, según esta invención, en materia prima de un proceso de obtención de hidrolizado de colágeno de baja acidez total.In the particular case of tissues subjected to prior extraction, these will be by-products of processes to obtain gelatins. The classic gelatin extraction processes are based, in broad strokes, on the maceration of the dermal tissue in conditions of extreme acidity or alkalinity for a total time of 4 to 24 hours, followed by a wash with demineralized water, a decantation of the fat released and neutralization with inorganic acid or basic solutions. Subsequently, the gelatin is solubilized, by extracting the solid tissue with aqueous solvents, in our particular case, the solvent is demineralized water. At this point, the gelatin solution is separated from the solid residue. The aqueous fraction is subjected to selective precipitation of the non-collagenic proteins, elimination of these by filtration, demineralization, drying and milling. The solid fraction, which in normal conditions would represent an important problem, due to its high organic matter load, becomes, according to this invention, the raw material of a process for obtaining collagen hydrolyzate of low total acidity.
Con Ia finalidad de obtener un hidrolizado de colágeno con unas caracterís- ticas que Io hagan apto para Ia elaboración de productos lácteos, Ia materia prima descrita anteriormente es sometida a una pluralidad de etapas hasta obtener un hidrolizado enzimático de colágeno de alta pureza, de peso molecular controlado y con unas características físico-químicas particulares y especiales que Io hacen apto para su uso en Ia elaboración de productos lácteos, especialmente aquellos que son sometidos a tratamientos térmicos.In order to obtain a collagen hydrolyzate with characteristics that make it suitable for the production of dairy products, the raw material described above is subjected to a plurality of stages until obtaining an enzymatic hydrolyzate of high purity collagen, of weight Molecular controlled and with particular and special physical-chemical characteristics that make it suitable for use in the production of dairy products, especially those that are subjected to heat treatments.
En Ia primera de las etapas, Ia materia prima de partida, tanto el tejido dérmico en forma nativa, como el subproducto de procesos de obtención de gelatina anteriormente descritos, se someten a una extracción acuosa a una temperatura comprendida entre 4O0C y 14O0C y durante un tiempo comprendido entre 2 y 8
horas. De un modo preferido, Ia extracción acuosa se realiza entre los 450C y los 1000C; y más preferiblemente entre los 5O0C y 8O0C. El tiempo de Ia extracción también puede variar y de un modo preferente suele tener una duración de 2 a 6 horas. Seguidamente, el pH de Ia solución extraída se ajusta entre 6,5 y 8,0, empleándose soluciones de ácidos o bases minerales. A tal fin, pueden emplearse por ejemplo soluciones de ácido clorhídrico. De un modo preferido y más preciso el pH se ajusta entre 7,4 y 7,6.In the first of the stages, the starting raw material, both the dermal tissue in a native form, and the byproduct of gelatin production processes described above, are subjected to an aqueous extraction at a temperature between 4O 0 C and 14O 0 C and for a time between 2 and 8 hours. In a preferred manner, the aqueous extraction is performed between 45 0 C and 100 0 C; and more preferably between 5O 0 C and 8O 0 C. The time of the extraction can also vary and preferably lasts from 2 to 6 hours. Then, the pH of the extracted solution is adjusted between 6.5 and 8.0, using solutions of acids or mineral bases. For this purpose, solutions of hydrochloric acid can be used, for example. In a preferred and more precise way the pH is adjusted between 7.4 and 7.6.
Una vez ajustado el pH, Ia solución es filtrada por ejemplo mediante el em- pleo de filtros de vidrio poroso, membranas de acetato de polivinilo, fibras de nylon, carbón activado o filtros rotativos de tierras de diatomeas. Evidentemente, pueden emplearse combinaciones de todos estos filtros hasta obtener un producto transparente.Once the pH has been adjusted, the solution is filtered, for example, by the use of porous glass filters, polyvinyl acetate membranes, nylon fibers, activated carbon or rotary diatomaceous earth filters. Obviously, combinations of all these filters can be used until a transparent product is obtained.
Dicha solución filtrada es sometida a una posterior etapa de desmineraliza- ción haciéndose pasar a través de resinas de intercambio iónico o equipos de diálisis. Para ello, se emplean, resinas mixtas, por ejemplo, de intercambio aniónico seguidas de intercambio catiónico o viceversa. Con esta etapa se consigue desmineralizar Ia solución filtrada hasta obtener una nueva solución con una conductividad absoluta inferior a 1.000 μS/cm y un contenido total inferior al 0,5% de sales minerales, o al menos un contenido en peso de sales minerales inferior al 1,0%, de modo que Ia riqueza en colágeno sobre extracto seco esté comprendida entre el 99% y el 99,5% en peso.Said filtered solution is subjected to a subsequent demineralization stage by being passed through ion exchange resins or dialysis equipment. For this, mixed resins are used, for example, anion exchange followed by cation exchange or vice versa. With this step it is possible to demineralize the filtered solution until obtaining a new solution with an absolute conductivity of less than 1,000 μS / cm and a total content of less than 0.5% of mineral salts, or at least a content of mineral salts by weight less than 1.0%, so that the richness in collagen on dry extract is between 99% and 99.5% by weight.
Los parámetros a controlar en esta etapa de desmineralización son Ia cantidad de solución a desmineralizar, que debe estar comprendida entre los 20 y los 300 litros por kilogramo de resina, siendo preferente el intervalo de 80 a 120 litros de solución por kilogramo de resina. Otro parámetro importante es Ia velocidad de paso de Ia solución a desmineralizar, Ia cual debe estar comprendida entre los 5 Htros/minuto/Kg de resina y los 100 litros/minuto/Kg de resina. Preferentemente Ia velocidad empleada es de 20 a 40 litros/minuto/Kg de resina. En el caso particular del uso de equipos de diálisis, Ia solución de colágeno, previamente filtrada, será sometida a sucesivos pasos a través de las membranas de diálisis, en configuración de recirculación con purga, hasta alcanzar un régimen estacionario en el que Ia conductividad absoluta de Ia solución no supere los 1.000 μS/cm.The parameters to be controlled in this demineralization stage are the amount of solution to be demineralized, which must be between 20 and 300 liters per kilogram of resin, with a range of 80 to 120 liters of solution per kilogram of resin being preferred. Another important parameter is the speed of the solution to be demineralized, which must be between 5 Htros / minute / Kg of resin and 100 liters / minute / Kg of resin. Preferably, the speed used is 20 to 40 liters / minute / kg of resin. In the particular case of the use of dialysis equipment, the collagen solution, previously filtered, will be subjected to successive steps through the dialysis membranes, in purge recirculation configuration, until a stationary regime is reached in which the absolute conductivity of the solution does not exceed 1,000 μS / cm.
Después de Ia etapa de desmineralización, Ia solución es ultrafiltrada me-
diante el paso por membranas con un diámetro de poro de 1.000 a 40.000 Daltons. Con esta etapa se pretenden eliminar todas las fracciones de restos proteicos de bajo peso molecular, así como todas aquellas sales minerales y aminas biógenas que son las causantes de males olores y mal sabor, tal como Ia putrescina o Ia cadaverina. De un modo preferente, las membranas utilizadas para Ia etapa de ul- trafiltración tienen un tamaño de poro de 20.000 Daltons.After the demineralization stage, the solution is ultrafiltered by means of through the passage through membranes with a pore diameter of 1,000 to 40,000 Daltons. With this stage, it is intended to eliminate all fractions of low molecular weight protein residues, as well as all those mineral salts and biogenic amines that are the cause of bad odors and bad taste, such as putrescine or cadaverine. Preferably, the membranes used for the ultrafiltration stage have a pore size of 20,000 Daltons.
Tras Ia ultrafiltración, Ia solución resultante es sometida a hidrólisis enzimáti- ca con enzimas proteolíticos bajo unas condiciones de temperatura y pH adecuados para que dichas enzimas puedan actuar. A modo de ejemplo pueden emplearse enzimas como Ia papaína, Ia ficina oAfter ultrafiltration, the resulting solution is subjected to enzymatic hydrolysis with proteolytic enzymes under adequate temperature and pH conditions so that said enzymes can act. By way of example, enzymes such as papain, ficin or
Ia bromelaína solas o en combinación.Bromelain alone or in combination.
Es también objeto de Ia presente invención las especiales condiciones de Ia hidrólisis enzimática. Para Ia obtención de un hidrolizado de colágeno de baja acidez y peso molecular entre 1.000 y 50.000 Daltons, preferentemente entre 5.000 y 20.000 Daltons se emplean dosis controladas de enzimas proteolíticos, a temperatura y tiempo controlados y pH constante. Las cantidades de enzima serán del orden de 1.000 Ul a 4.000 Ul por kilogramo de producto seco a hidrolizar. Se ha observado que concentraciones de enzima comprendidas entre 1.500 Ul y 2.500 Ul por kilogramo de producto seco son preferidas para Ia obtención de hidrolizados de colágeno de baja acidez total.The special conditions of enzymatic hydrolysis are also object of the present invention. To obtain a collagen hydrolyzate of low acidity and molecular weight between 1,000 and 50,000 Daltons, preferably between 5,000 and 20,000 Daltons, controlled doses of proteolytic enzymes are used, at controlled temperature and time and constant pH. The amounts of enzyme will be of the order of 1,000 Ul to 4,000 Ul per kilogram of dry product to be hydrolyzed. It has been observed that enzyme concentrations between 1,500 Ul and 2,500 Ul per kilogram of dry product are preferred for obtaining collagen hydrolysates of low total acidity.
Las temperaturas de trabajo, así como los tiempos de hidrólisis preferidos dependerán del enzima, o combinación de enzimas, utilizado. A su vez, el pH se controlará durante toda Ia hidrólisis para evitar una deriva superior a 2 unidades respecto al pH óptimo de actuación del enzima, o combinación de enzimas, prefe- rentemente se mantendrá un pH, durante toda Ia hidrólisis, dentro de un rango de ±0,5 unidades respecto al pH óptimo.Working temperatures, as well as preferred hydrolysis times will depend on the enzyme, or combination of enzymes, used. In turn, the pH will be controlled throughout the hydrolysis to avoid a drift greater than 2 units with respect to the optimum pH of the enzyme, or combination of enzymes, preferably a pH will be maintained, during the entire hydrolysis, within a range of ± 0.5 units with respect to the optimum pH.
El rango de temperaturas a las que se realiza Ia etapa de hidrólisis enzimática suele estar comprendido entre los 150C y los 8O0C, en función del rango óptimo de trabajo de Ia o las enzimas. Se obtienen resultados muy satisfactorios en un rango de temperaturas de los 4O0C a los 7O0C. Las enzimas se dejan actuar sobre Ia solución ultrafiltrada durante de 30 a 200 minutos, preferentemente de 90 a 120 minutos y a un pH de Ia solución de reacción entre 5 y 9, más concretamente entre 6,8 y 7,8, aunque este valor dependerá de los enzimas utilizados.The range of temperatures at which the stage of enzymatic hydrolysis is carried out is usually between 15 0 C and 8O 0 C, depending on the optimum working range of Ia or enzymes. Very satisfactory results are obtained in a temperature range of 4O 0 C to 7O 0 C. The enzymes are allowed to act on the ultrafiltered solution for 30 to 200 minutes, preferably 90 to 120 minutes and at a pH of the solution of reaction between 5 and 9, more specifically between 6.8 and 7.8, although this value will depend on the enzymes used.
Para una correcta evolución de Ia hidrólisis, se realiza un control continuo
del pH y se compensa Ia deriva natural de este parámetro mediante Ia adición de soluciones diluidas de álcali, por ejemplo solución de hidróxido sódico.For a correct evolution of the hydrolysis, a continuous control is carried out of the pH and the natural drift of this parameter is compensated by the addition of diluted alkali solutions, for example sodium hydroxide solution.
Una vez transcurrido el tiempo fijado para Ia hidrólisis enzimática deben desactivarse las enzimas, que al tratarse generalmente de enzimas termolábiles es factible mediante el solo hecho de aumentar Ia temperatura del producto a temperaturas de 90° C entre 5 y 30 minutos.Once the time set for enzymatic hydrolysis has elapsed, the enzymes must be deactivated, since they are generally thermolabile enzymes, it is only possible by increasing the temperature of the product at temperatures of 90 ° C between 5 and 30 minutes.
La solución de hidrolizado enzimático de colágeno obtenida es sometida a una nueva etapa de filtración con Ia finalidad de asegurar Ia ausencia de cualquier tipo de partículas que pudieran inestabilizar el producto, así como dar una transpa- rencia total al mismo. A modo de ejemplo, el filtro a emplear puede ser de marcos y placas, de filtración en profundidad o bien filtros de vidrio poroso.The collagen enzymatic hydrolyzate solution obtained is subjected to a new filtration stage in order to ensure the absence of any type of particles that could destabilize the product, as well as give a total transparency to it. As an example, the filter to be used can be frames and plates, deep filtration or porous glass filters.
En cualquier caso, esta etapa de filtración de Ia solución de hidrolizado enzimático de colágeno va seguida de una etapa de ultrafiltración a través de membranas con un diámetro de poro de 1.000 a 40.000 Daltons. De hecho, el objetivo de esta ultrafiltración es el de eliminar todas aquellas fracciones de péptidos superiores a 20.000 Daltons, así como las fracciones de muy bajo peso molecular (< 1.000 Daltons). En realidad, esta etapa de ultrafiltración consiste en una filtración o exclusión molecular para seleccionar todas aquellas fracciones de hidrolizado enzimático de colágeno, especialmente las de peso molecular comprendido entre los 5.000 Daltons y los 20.000 Daltons.In any case, this stage of filtration of the collagen enzyme hydrolyzate solution is followed by an ultrafiltration stage through membranes with a pore diameter of 1,000 to 40,000 Daltons. In fact, the objective of this ultrafiltration is to eliminate all those peptide fractions greater than 20,000 Daltons, as well as very low molecular weight fractions (<1,000 Daltons). In reality, this ultrafiltration stage consists of a molecular filtration or exclusion to select all those collagen enzyme hydrolyzate fractions, especially those of molecular weight between 5,000 Daltons and 20,000 Daltons.
Llegados a este punto, Ia solución de hidrolizado enzimático de colágeno es concentrada mediante calefacción a temperaturas inferiores o iguales a 7O0C con el objetivo de obtener un producto concentrado que pueda ser secado por ejemplo por atomización. De un modo preferente, Ia etapa de concentración se lleva a cabo a una temperatura inferior o igual a 6O0C y mediante Ia aplicación de vacío a fin de evaporar el agua a temperaturas por debajo de Ia temperatura de ebullición de esta última. Así, se suelen emplear presiones que oscilan entre los 10 y los 0,0001 mm de Hg, preferentemente de 1,0 a 0,01 mm de Hg. Dicha etapa de concentración se da por finalizada cuando se alcanzan concentraciones finales de hidrolizado enzimático de colágeno en solución del 15% al 45% en peso. Concretamente, es preferido un rango de concentraciones del 30% al 45% en peso de hidrolizado enzimático de colágeno en solución.
Finalmente, la solución de hidrolizado enzimático de colágeno con un porcentaje en peso del mismo del 30% al 45% es secada hasta obtener un producto seco con una humedad inferior o igual al 8% y una densidad comprendida entre los 0,3 g/ml y 0,8 g/ml, preferentemente entre 0,4 g/ml y 0,6 g/ml. Esta etapa final de secado del producto se realiza por liofilización o bien por atomización.At this point, the collagen enzyme hydrolyzate solution is concentrated by heating at temperatures below or equal to 7O 0 C in order to obtain a concentrated product that can be dried, for example, by atomization. Preferably, the concentration stage is carried out at a temperature lower than or equal to 6O 0 C and by means of the application of vacuum in order to evaporate the water at temperatures below the boiling temperature of the latter. Thus, pressures ranging from 10 to 0.0001 mm Hg are used, preferably from 1.0 to 0.01 mm Hg. Said concentration step is terminated when final concentrations of collagen enzyme hydrolyzate in solution of 15% to 45% by weight are reached. Specifically, a concentration range of 30% to 45% by weight of collagen enzyme hydrolyzate in solution is preferred. Finally, the collagen enzyme hydrolyzate solution with a weight percentage of 30% to 45% is dried until a dry product is obtained with a humidity less than or equal to 8% and a density between 0.3 g / ml and 0.8 g / ml, preferably between 0.4 g / ml and 0.6 g / ml. This final stage of drying the product is carried out by lyophilization or by atomization.
El procedimiento según se ha descrito permite obtener de un modo económico un hidrolizado enzimático de colágeno con parámetros constantes de lote a lote y altamente estable. Los estudios de estabilidad realizados han revelado que el citado hidrolizado tiene un periodo de validez de unos 4 años almacenado a una temperatura controlada de 250C y a una humedad relativa del ambiente inferior al 60%.The process as described allows to obtain in an economic way an enzymatic hydrolyzate of collagen with constant parameters from batch to batch and highly stable. Stability studies have revealed that the said hydrolyzate has a shelf life of about 4 years when stored at a controlled temperature of 25 0 C and a relative humidity of less than 60% atmosphere.
El hidrolizado enzimático de colágeno obtenible de acuerdo con el procedimiento anteriormente descrito, se caracteriza por su gran pureza y por sus caracte- rísticas organolépticas idóneas para ser administrado en preparaciones orales. Concretamente, el hidrolizado enzimático de colágeno comprende un 99% en peso de hidrolizado sobre producto seco.The collagen enzymatic hydrolyzate obtainable according to the procedure described above, is characterized by its high purity and its ideal organoleptic characteristics to be administered in oral preparations. Specifically, the enzymatic collagen hydrolyzate comprises 99% by weight of hydrolyzate on dry product.
Dicho hidrolizado enzimático de colágeno, también objeto de Ia invención, tiene un peso molecular comprendido entre los 1.000 Daltons y los 50.000 Daltons, siendo el rango preferido de pesos moleculares de 5.000 a 20.000 Daltons.Said collagen enzyme hydrolyzate, also object of the invention, has a molecular weight between 1,000 Daltons and 50,000 Daltons, the preferred molecular weight range of 5,000 to 20,000 Daltons.
Los parámetros controlados en el producto final y sus límites se exponen a continuación:The parameters controlled in the final product and their limits are set out below:
Riqueza: Debe ser superior al 99% sobre producto seco.Wealth: Must be greater than 99% on dry product.
Humedad: Debe ser inferior al 8% Densidad aparente: Debe estar comprendida entre 0,3 y 0,8 g/ml, de preferencia 0,4 y 0,6 g/ml. pH (sol. al 20%): Debe estar entre 6,0 y 7,0Humidity: It must be less than 8%. Bulk density: It must be between 0.3 and 0.8 g / ml, preferably 0.4 and 0.6 g / ml. pH (20% sol.): Must be between 6.0 and 7.0
Acidez (grados Dornic): Inferior a 250D.Acidity (Dornic degrees): Less than 25 0 D.
Viscosidad (sol. al 20%): Debe estar comprendida entre 15 y 25 mps Turbidez (sol. al 20%): Debe ser inferior a 25 NTUViscosity (20% sol.): Must be between 15 and 25 mps Turbidity (20% sol.): Must be less than 25 NTU
Conductividad relativa (sol. al 1%): Debe ser inferior a 500 μS/cmRelative conductivity (1% sol.): Must be less than 500 μS / cm
Cenizas: Deben ser inferiores al 1%Ashes: Must be less than 1%
Las características de salinidad, baja acidez y pesos moleculares del hidrolizado enzimático de acuerdo con Ia invención, Io hacen adecuado y óptimo para su
aplicación en productos lácteos o que contengan leche, sin que por ello se produzca Ia degradación de dicho producto lácteo o se vean modificadas sus características físicas y/o organolépticas.The characteristics of salinity, low acidity and molecular weights of the enzymatic hydrolyzate according to the invention, make it suitable and optimal for its application in dairy products or those containing milk, without this causing degradation of said dairy product or its physical and / or organoleptic characteristics are modified.
A continuación se detallan a modo de ejemplos no limitativos, variantes del procedimiento de obtención de hidrolizado de colágeno según Ia invención. También se detallan los parámetros controlados en el hidrolizado enzimático obtenible por el procedimiento.Below are detailed by way of non-limiting examples, variants of the process for obtaining collagen hydrolyzate according to the invention. The controlled parameters in the enzymatic hydrolyzate obtainable by the process are also detailed.
EJEMPLO 1 7.500 Kg de tejido dérmico de origen porcino, obtenido directamente, y en condiciones higiénicas, de sala de despiece, y conservado a una temperatura inferior a 5o C, se someten a las siguientes etapas:EXAMPLE 1 7,500 Kg of dermal tissue of porcine origin, obtained directly, and in hygienic conditions, from cutting plant, and kept at a temperature below 5 o C, are subjected to the following stages:
1.- Maceración en solución de ácido clorhídrico durante 4 horas. 2.- Neutralización a pH superior a 4 e inferior a 6 con hidróxido sódico.1.- Maceration in hydrochloric acid solution for 4 hours. 2.- Neutralization at pH higher than 4 and lower than 6 with sodium hydroxide.
3.- Extracción a 60° C durante 4 horas. 4.- Ajuste de pH a 7,5 empleando hidróxido sódico. 5.- Filtración con filtro rotativo de tierra de diatomeas. 6.- Desmineralización mediante membranas de diálisis, en configuración de recirculación y purga, hasta un régimen estacionario en el que Ia conductividad absoluta no supere los 1.000 μS/cm. 7.- Ultrafiltración a través de una membrana de 20.000 Daltons. 8.- Hidrólisis controlada enzimáticamente añadiendo 1.350.000 U. I. de en- dopeptidasa de origen bacteriano sobre el producto a pH 7,2 a 60° C duran- te 110 minutos.3.- Extraction at 60 ° C for 4 hours. 4.- Adjust pH to 7.5 using sodium hydroxide. 5.- Filtration with rotary diatomaceous earth filter. 6.- Demineralization by dialysis membranes, in recirculation and purge configuration, until a steady state in which the absolute conductivity does not exceed 1,000 μS / cm. 7.- Ultrafiltration through a membrane of 20,000 Daltons. 8.- Enzymatically controlled hydrolysis by adding 1,350,000 U. I. of bacterial origin dopeptidase on the product at pH 7.2 at 60 ° C for 110 minutes.
9.- Desactivación de los enzimas mediante tratamiento térmico a 90° C durante 20 minutos.9.- Deactivation of enzymes by heat treatment at 90 ° C for 20 minutes.
10.- Filtración a través de filtro de marcos y placas de 25 μm. 11.- Ultrafiltración a través de membrana de 5.000 Daltons. 12.- Concentración a 45° C con un vacío de 0,15 mm de Hg hasta una concentración de sólidos del 40 % 13.- Secado por atomización (spray drying)
El producto (hidrolizado enzimático de colágeno) obtenido con este proceso (805 Kg) presentó las siguientes características físico químicas:10.- Filtration through 25 μm frame and plate filter. 11.- Ultrafiltration through 5,000 Daltons membrane. 12.- Concentration at 45 ° C with a vacuum of 0.15 mm of Hg up to a solids concentration of 40% 13.- Spray drying The product (collagen enzyme hydrolyzate) obtained with this process (805 Kg) had the following physical chemical characteristics:
Riqueza (sobre producto seco): 99,5 % Humedad: 4,5 %Wealth (on dry product): 99.5% Humidity: 4.5%
Densidad aparente: 0,483 g/ml pH (sol. al 20%): 6,08Bulk density: 0.483 g / ml pH (20% sol): 6.08
Acidez (10 g): 19,80D. (Según Ia escala Dornic) Viscosidad (sol. al 20%): 20,9 mps Turbidez (sol. al 20%): 24,6 NTUAcidity (10 g): 19.8 0 D. (According to the Dornic scale) Viscosity (20% sol.): 20.9 mps Turbidity (20% sol.): 24.6 NTU
Conductividad relativa (0,5%): 315 μS/cm Cenizas: 0,5 %Relative conductivity (0.5%): 315 μS / cm Ash: 0.5%
EJEMPLO 2 7.500 Kg de tejido dérmico de origen porcino, obtenido directamente, y en condiciones higiénicas, de sala de despiece, y conservado a una temperatura inferior a 5o C, se someten a las siguientes etapas:EXAMPLE 2 7,500 Kg of dermal tissue of porcine origin, obtained directly, and in hygienic conditions, from cutting plant, and kept at a temperature below 5 o C, are subjected to the following stages:
1.- Maceración en solución de ácido clorhídrico durante 4 horas. 2.- Neutralización a pH superior a 4 e inferior a 6 con hidróxido sódico.1.- Maceration in hydrochloric acid solution for 4 hours. 2.- Neutralization at pH higher than 4 and lower than 6 with sodium hydroxide.
3.- Extracción a 65° C durante 4 horas. 4.- Ajuste de pH a 7,5 empleando hidróxido sódico. 5.- Filtración con filtro rotativo de tierra de diatomeas. 6.- Desmineralización mediante membranas de diálisis, en configuración de recirculación y purga, hasta un régimen estacionario en el que Ia conductividad absoluta no supere los 1.000 μS/cm. 7.- Ultrafiltración a través de una membrana de 20.000 Daltons. 8.- Hidrólisis controlada enzimáticamente añadiendo 2.000.000 U. I. de en- dopeptidasa de origen bacteriano sobre el producto a pH 7,8 a 60° C duran- te 110 minutos.3.- Extraction at 65 ° C for 4 hours. 4.- Adjust pH to 7.5 using sodium hydroxide. 5.- Filtration with rotary diatomaceous earth filter. 6.- Demineralization by dialysis membranes, in recirculation and purge configuration, until a steady state in which the absolute conductivity does not exceed 1,000 μS / cm. 7.- Ultrafiltration through a membrane of 20,000 Daltons. 8.- Enzymatically controlled hydrolysis by adding 2,000,000 U. I. of bacterial origin dopeptidase on the product at pH 7.8 at 60 ° C for 110 minutes.
9.- Desactivación de los enzimas mediante tratamiento térmico a 90° C durante 20 minutos.9.- Deactivation of enzymes by heat treatment at 90 ° C for 20 minutes.
10.- Filtración a través de filtro de marcos y placas de 10 μm. 11.- Ultrafiltración a través de membrana de 5.000 Daltons.
12.- Concentración a 40° C con un vacío de 0,1 mm de Hg hasta una concentración de sólidos del 45 %. 13.- Secado por atomización (spray drying)10.- Filtration through 10 μm frame and plate filter. 11.- Ultrafiltration through 5,000 Daltons membrane. 12.- Concentration at 40 ° C with a vacuum of 0.1 mm Hg to a solids concentration of 45%. 13.- Spray drying
El producto obtenido (hidrolizado enzimático de colágeno) con este procesoThe product obtained (enzymatic collagen hydrolyzate) with this process
(950 Kg) presentó las siguientes características físico químicas:(950 Kg) presented the following physical chemical characteristics:
Riqueza (sobre producto seco): 99,6 %Wealth (on dry product): 99.6%
Humedad: 4,9 % Densidad aparente: 0,495 g/ml pH (sol. al 20%): 6,12Humidity: 4.9% Bulk density: 0.495 g / ml pH (20% sol.): 6.12
Acidez (10 g): 16,9 0D. (Según Ia escala Dornic)Acidity (10 g): 16.9 0 D. (According to the Dornic scale)
Viscosidad (sol. al 20%): 21 ,1 mpsViscosity (20% sol.): 21, 1 mps
Turbidez (sol. al 20%): 18,5 NTU Conductividad relativa (0,5%): 210 μS/cmTurbidity (20% sol.): 18.5 NTU Relative conductivity (0.5%): 210 μS / cm
Cenizas: 0,4 %Ashes: 0.4%
Evidentemente, el hidrolizado enzimático de colágeno de acuerdo con Ia invención con una acidez total, expresada en grados Dornic, inferior a 25°D; y una conductividad en solución al 1% no superior a 500 μS/cm, puede obtenerse mediante otros procedimientos, tal como procedimientos biotecnológicos en los cuales, puede pensarse en Ia inmovilización de las enzimas en soportes extraíbles y fácilmente recuperables, o mediante el aporte de las enzimas de interés trabajando "in situ" dentro de unidades celulares por ejemplo bacterias.Obviously, the enzymatic collagen hydrolyzate according to the invention with a total acidity, expressed in Dornic degrees, less than 25 ° D; and a conductivity in 1% solution not exceeding 500 μS / cm, can be obtained by other procedures, such as biotechnological procedures in which, the immobilization of enzymes on removable and easily recoverable supports, or by the contribution of the enzymes of interest working "in situ" within cell units for example bacteria.
Ejemplo de las evidencias obtenidas mediante Ia aplicación del procedimiento de acuerdo con Ia invención expuesto anteriormente:Example of the evidence obtained by applying the procedure according to the invention set forth above:
En las condiciones anteriormente detalladas, se obtiene un hidrolizado de colágeno de baja acidez siendo así compatibles con Ia fabricación de suplementos nutricionales en los que se comprende como ingrediente principal Ia leche, o derivados lácteos, y en los que el hidrolizado de colágeno está presente en concentraciones comprendidas entre el 2% y el 15% en peso, sin superar una acidez total de 50° en Ia escala Dornic. En estas condiciones las mezclas de hidrolizado de colágeno y leche, o derivados lácteos, en las dosis detalladas anteriormente, son esta-
bles a los procesos de pasteurización y esterilización, y a Ia fermentación posterior, en Ia producción de suplementos alimenticios basados en leche, o derivados lácteos, enriquecidos con hidrolizado enzimático de colágeno.Under the conditions detailed above, a low acidity collagen hydrolyzate is obtained, thus being compatible with the manufacture of nutritional supplements in which milk, or milk derivatives, is comprised as the main ingredient, and in which the collagen hydrolyzate is present in concentrations between 2% and 15% by weight, without exceeding a total acidity of 50 ° in the Dornic scale. Under these conditions the mixtures of collagen and milk hydrolyzate, or milk derivatives, in the doses detailed above, are stable. The process of pasteurization and sterilization, and subsequent fermentation, in the production of food supplements based on milk, or dairy products, enriched with enzymatic collagen hydrolyzate.
A modo de ejemplo se presentan en las Figs. 1 y 2 dos cromatogramas co- rrespondientes a Ia distribución de pesos moleculares de sendos hidrolizados de colágeno, el primero (Rg. 1) corresponde a un hidrolizado de colágeno preparado según el proceso clásico, el segundo (Fig. 2) corresponde al preparado según las condiciones de proceso detalladas anteriormente. También se citan los valores de acidez correspondientes, expresados en grados de Ia escala Dornic. Se puede ob- servar que se ha conseguido una acidez compatible con Ia utilización del hidrolizado de colágeno en Ia formulación de productos lácteos pasteurizados y fermentados, a Ia vez que se conserva el perfil original de pesos moleculares. La reducción de Ia acidez es del 38%, mientras se observa que el perfil de pesos moleculares es equivalente. Así, en Ia Fig. 1 aparece representado el cromatograma de un hidrolizado enzimático de colágeno, preparado por el procedimiento según el estado de Ia técnica, con una acidez total de 36,37 "Dornic.As an example, they are presented in Figs. 1 and 2 two chromatograms corresponding to the molecular weight distribution of two collagen hydrolysates, the first (Rg. 1) corresponds to a collagen hydrolyzate prepared according to the classical process, the second (Fig. 2) corresponds to the preparation according to the process conditions detailed above. The corresponding acidity values are also mentioned, expressed in degrees of the Dornic scale. It can be observed that acidity compatible with the use of collagen hydrolyzate in the formulation of pasteurized and fermented milk products has been achieved, while maintaining the original molecular weight profile. The reduction in acidity is 38%, while observing that the molecular weight profile is equivalent. Thus, in Fig. 1 the chromatogram of an enzymatic collagen hydrolyzate, prepared by the method according to the state of the art, with a total acidity of 36.37 "Dornic, is represented.
Alternativamente, en Ia Fig. 2, que corresponde a un hidrolizado de colágeno según Ia invención y obtenido por el procedimiento de acuerdo con Ia invención, cuya acidez total es de 22,56 °Dornic.Alternatively, in Fig. 2, which corresponds to a collagen hydrolyzate according to the invention and obtained by the process according to the invention, whose total acidity is 22.56 ° Dornic.
Ejemplos de alimentos funcionales v suplementos nutricionales: De acuerdo con otro de los objetos de Ia presente invención, a continuación se detallan ejemplos de alimentos funcionales y suplementos nutricionales que comprenden el hidrolizado enzimático de colágeno de baja acidez según Ia invención, obtenible de acuerdo con el procedimiento anteriormente descrito.Examples of functional foods and nutritional supplements: According to another of the objects of the present invention, examples of functional foods and nutritional supplements comprising the enzymatic hydrolyzate of low acid collagen according to the invention, obtainable according to the invention, are detailed below. procedure described above.
Estos alimentos funcionales o suplementos nutricionales pueden comprender además un compuesto seleccionado independientemente de entre: compuestos antioxidantes, sales minerales, vitaminas y/o coenzimas, extractos vegetales, ex- tractos de origen animal y compuestos coadyuvantes del hidrolizado enzimático de colágeno. También puede emplearse una mezcla de todos ellos.
EJEMPLO 1: Alimento funcional: yogurtThese functional foods or nutritional supplements may further comprise a compound independently selected from: antioxidant compounds, mineral salts, vitamins and / or coenzymes, plant extracts, extracts of animal origin and adjuvant compounds of the collagen enzyme hydrolyzate. A mixture of all of them can also be used. EXAMPLE 1: Functional food: yogurt
Se ha preparado con buenos resultados un yogurt con Ia siguiente composición:A yogurt with the following composition has been prepared with good results:
Leche 73,9 g Hidrolizado enzimático de colágeno 10,O gMilk 73.9 g Collagen Enzymatic Hydrolyzate 10, O g
Leche en polvo desnatada 12,0 gSkimmed milk powder 12.0 g
Nata 4,0 gCream 4.0 g
Fermentos lácticos 0,1 g0.1 g lactic ferments
El sabor del yogurt es agradable así como su aspecto y textura.The taste of yogurt is pleasant as well as its appearance and texture.
EJEMPLO 2: Alimento funcional: Postre de chocolateEXAMPLE 2: Functional food: Chocolate dessert
Se ha preparado un postre de chocolate con Ia siguiente composición:A chocolate dessert with the following composition has been prepared:
Leche desnatada 34,8 gSkim milk 34.8 g
Hidrolizado enzimático de colágeno 10,0 g Azúcar 15,O gCollagen Enzymatic Hydrolyzate 10.0 g Sugar 15, O g
Lactosa 10,0 gLactose 10.0 g
Nata 5,0 gCream 5.0 g
Almidón de maíz pregelatinizado 12,O gPregelatinized corn starch 12, O g
Cacao desgrasado 10,0 g Carragenato 2,O gDefatted cocoa 10.0 g Carrageen 2, O g
Gelatina 1,0 g1.0 g jelly
Emulgente 0,2 g0.2 g emulsifier
Tanto Ia extensibilidad como Ia viscosidad, Ia textura y el sabor no se ven alterados mientras mejoramos Ia capacidad nutricional del alimento.Both the extensibility and the viscosity, texture and taste are not altered while improving the nutritional capacity of the food.
EJEMPLO 3: Alimento funcional: yogurt bebibleEXAMPLE 3: Functional food: drinkable yogurt
Se ha preparado un yogurt bebible con Ia siguiente composición:A drinkable yogurt has been prepared with the following composition:
Leche 81,4 gMilk 81.4 g
Hidrolizado enzimático de colágeno 10,0 g Leche en polvo desnatada 6,0 gCollagen Enzymatic Hydrolyzate 10.0 g Skimmed milk powder 6.0 g
Nata 2,5 gCream 2.5 g
Fermentos lácticos 0,1 g0.1 g lactic ferments
Esta preparación admite distintas saborizaciones para obtener una gama de yogures bebibles con propiedades funcionales.
EJEMPLO 4: Alimento funcional: Batido de leche v chocolate Se ha preparado un batido de leche y chocolate sin que Ia adición del hidro- lizado enzimático de colágeno modifique las características organolépticas del mismo. Leche desnatada 36,O gThis preparation allows different flavorings to obtain a range of drinkable yogurts with functional properties. EXAMPLE 4: Functional food: Milk and chocolate milkshake A milk and chocolate milkshake has been prepared without the addition of the enzymatic collagen hydrolyzate modifying its organoleptic characteristics. Skim milk 36, O g
Sacarosa 18,0 gSucrose 18.0 g
Proteínas de leche 12,0 gMilk proteins 12.0 g
Hidrolizado enzimático de colágeno 6,6 gEnzymatic collagen hydrolyzate 6.6 g
Cacao en polvo 6,0 g Almidón de maíz 5,4 gCocoa powder 6.0 g Corn starch 5.4 g
Aceite de soja 5,0 g5.0 g soybean oil
Fibra vegetal 5,0 gVegetable fiber 5.0 g
Carragenatos 2,0 gCarrageenans 2.0 g
Fosfato disódico 1,5 g Diglicéridos de ácidos grasos 1,0 gDisodium phosphate 1.5 g Fatty acid diglycerides 1.0 g
Cloruro sódico 0,5 g0.5 g sodium chloride
Complejo de minerales 0,5 gMineral complex 0.5 g
Complejo vitamínico (A, D, C, grupo B) 0,5 gVitamin complex (A, D, C, group B) 0.5 g
EJEMPLO 5: Alimento funcional: Queso fundidoEXAMPLE 5: Functional food: Molten cheese
Queso blanco 42,5 gWhite Cheese 42.5 g
Crema de leche 30,O gMilk cream 30, O g
Proteínas de leche 10,0 g10.0 g milk proteins
Hidrolizado enzimático de colágeno 10,0 g Fosfato calcico 2,O gCollagen Enzymatic Hydrolyzate 10.0 g Calcium Phosphate 2, O g
Polifosfatos 1,5 g1.5 g polyphosphates
Fosfato disódico 1,5 g1.5 g disodium phosphate
Citrato sódico 1,0 g1.0 g sodium citrate
Cloruro sódico 1,0 g Concentrado de minerales de leche 0,5 gSodium Chloride 1.0 g Milk Mineral Concentrate 0.5 g
La adición del hidrolizado de colágeno no modifica de manera apreciable el sabor, Ia extensibilidad ni Ia cremosidad del producto.
EJEMPLO 6: Complemento nutrícional: Barritas energéticasThe addition of collagen hydrolyzate does not appreciably modify the taste, extensibility or creaminess of the product. EXAMPLE 6: Nutritional Supplement: Energy Bars
Sacarosa 28,0 gSucrose 28.0 g
Manteca de cacao 25,O gCocoa butter 25, O g
Leche entera en polvo 18,O g Hidrolizado enzimático de Colágeno 8,0 gWhole milk powder 18, O g Collagen enzymatic hydrolyzate 8.0 g
Harina de trigo 4,5 gWheat Flour 4.5 g
Pasta de cacao 4,8 gCocoa Paste 4.8 g
Harina de maíz 3,5 g3.5 g cornmeal
Salvado de trigo 3,0 g Dextrina de maíz 2,O gWheat bran 3.0 g Corn dextrin 2, O g
Cloruro sódico 1,0 g1.0 g sodium chloride
Lecitina 0,5 g0.5 g lecithin
Fructosa 1,0 gFructose 1.0 g
Complejo vitamínico (C, D, grupo B) 0,5 g Aromas 0,2 gVitamin complex (C, D, group B) 0.5 g Aromas 0.2 g
Modificando algunos de los componentes es factible preparar barritas para ayudar al control del peso.By modifying some of the components it is feasible to prepare bars to help control the weight.
En todos estos ejemplos, se observa que para Ia elaboración de suplemen- tos nutricionales o alimentos funcionales en los que se incluye Ia leche, o un derivado lácteo, Ia acidez total del producto, expresada como Ia cantidad de hidróxido sódico consumido para Ia neutralización de 10 gramos de hidrolizado de colágeno, según Ia escala de unidades Dornic, debe ser inferior a 50. Preferiblemente inferior o igual a 40. Ensayo clínico para determinar Ia mejora en calidad de vida v capacidades funcionales en humanosIn all these examples, it is observed that for the elaboration of nutritional supplements or functional foods in which milk, or a dairy derivative, is included, the total acidity of the product, expressed as the amount of sodium hydroxide consumed for the neutralization of 10 grams of collagen hydrolyzate, according to the Dornic unit scale, must be less than 50. Preferably less than or equal to 40. Clinical trial to determine the improvement in quality of life and functional capabilities in humans
De acuerdo con otro de los objetos de Ia presente invención, los inventores han determinado un nuevo uso del hidrolizado enzimático de colágeno obtenible por el procedimiento anteriormente descrito. Concretamente, el hidrolizado enzimático de colágeno se emplea para Ia preparación de un suplemento nutricional o alimento funcional para mejorar Ia capacidad funcional y/o Ia calidad de vida de un mamífero, concretamente un humano, con dificultades funcionales derivadas de alteraciones de los cartílagos articulares.
Así, con el fin de estudiar Ia posible aplicación del nuevo hidrolizado enzimá- tico de colágeno de elevada pureza y de mejor biodisponiblidad (peso molecular inferior, entre 5.000 y 20.000 Daltons) como alimento funcional o como componente de alimentos funcionales o suplementos nutricionales, se decidió llevar a cabo un estudio en 100 voluntarios a los que a 50 de ellos se administraría este hidrolizado enzimático de colágeno en una dosis de 10 g diarios, equivalente a una dosis de entre 0,1 y 0,25 g por Kg de peso corporal de hidrolizado enzimático de colágeno y a los otros 50 una dosis diaria de 1 ,5 g de sulfato de glucosamina, equivalente a una dosis entre 0,015 y 0,0375 g por Kg de peso corporal. El sulfato de glucosami- na es ampliamente utilizado tanto en Europa como en Estados Unidos a estas dosis para el tratamiento de alteraciones articulares en general y varios estudios validan que Ia administración de glucosamina es un medio eficaz para mejorar el dolor y Ia movilidad de las articulaciones con disfunciones consecutivas a alteraciones de los cartílagos articulares. Con este ensayo se pretendía valorar el efecto del nuevo hidrolizado enzimático de colágeno en aquellas afecciones donde habitualmente se emplean los conocidos antiinflamatorios no esteroideos (AINES), concretamente en aquellos trastornos de Ia capacidad funcional o calidad de vida derivados de alteraciones de los cartílagos articulares. Por otra parte, el sulfato de glucosamina es también ampliamente utilizado en Europa y sobre todo en Estados Unidos como alimento funcional o como integrante de alimentos funcionales y suplementos nutricionales (Ia monografía de Ia USP 24 Io incluye en suplementos nutricionales) a estas mismas dosis (1,5 g diarios) y se asegura que mejoran en general las funciones corporales, principalmente las mecánicas (movilidad, agilidad ) y que mejoran el estado general y Ia calidad de vida.In accordance with another of the objects of the present invention, the inventors have determined a new use of the enzymatic collagen hydrolyzate obtainable by the process described above. Specifically, the enzymatic collagen hydrolyzate is used for the preparation of a nutritional supplement or functional food to improve the functional capacity and / or the quality of life of a mammal, specifically a human, with functional difficulties derived from alterations of the articular cartilage. Thus, in order to study the possible application of the new collagen enzyme hydrolyzate of high purity and of better bioavailability (lower molecular weight, between 5,000 and 20,000 Daltons) as a functional food or as a component of functional foods or nutritional supplements, decided to carry out a study in 100 volunteers to whom 50 of them would be given this collagen enzyme hydrolyzate in a dose of 10 g daily, equivalent to a dose between 0.1 and 0.25 g per kg of body weight of collagen enzyme hydrolyzate and the other 50 a daily dose of 1.5 g of glucosamine sulfate, equivalent to a dose between 0.015 and 0.0375 g per kg of body weight. Glucosamine sulfate is widely used both in Europe and in the United States at these doses for the treatment of joint disorders in general and several studies validate that the administration of glucosamine is an effective means to improve pain and joint mobility. with consecutive dysfunctions to alterations of articular cartilage. With this test it was intended to assess the effect of the new enzymatic collagen hydrolyzate in those conditions where the known non-steroidal anti-inflammatory drugs (NSAIDs) are usually used, specifically in those disorders of the functional capacity or quality of life derived from alterations of the articular cartilage. On the other hand, glucosamine sulfate is also widely used in Europe and especially in the United States as a functional food or as a member of functional foods and nutritional supplements (the USP 24 Io monograph includes nutritional supplements) at these same doses ( 1.5 g per day) and it is ensured that body functions generally improve, mainly mechanical ones (mobility, agility) and that they improve the general state and quality of life.
El ensayo clínico consistió en un estudio multicéntrico, doble ciego, paralelo y comparativo entre el nuevo hidrolizado enzimático de colágeno y el sulfato de glucosamina. La finalidad era comparar Ia mejoría en Ia calidad de vida (según el cuestionario genérico SF-36 (The MOS 36-item short-form health survey (SF-36). I. Conceptual framework and item selection. Med care 1992 Jun; 30 (6): 473-483), así como Ia eficacia, Ia seguridad y Ia tolerabilidad de ambos tratamientos administrados por vía oral durante 90 días consecutivos.
El estudio se llevó a cabo en 100 voluntarios (de los que fueron evaluables 93) afectos de alteraciones articulares que inducían dolor, disminución de Ia movilidad y alteración y pérdida notables de Ia calidad de vida.The clinical trial consisted of a multicenter, double-blind, parallel and comparative study between the new enzymatic collagen hydrolyzate and glucosamine sulfate. The purpose was to compare the improvement in quality of life (according to the generic SF-36 questionnaire (The MOS 36-item short-form health survey (SF-36). I. Conceptual framework and item selection. Med care 1992 Jun; 30 (6): 473-483), as well as the efficacy, safety and tolerability of both treatments administered orally for 90 consecutive days. The study was carried out in 100 volunteers (93 of whom were evaluable) affected by joint disorders that induced pain, decreased mobility and notable alteration and loss of quality of life.
Los sujetos fueron distribuidos en dos grupos de 50 individuos, de los que fueron evaluables 47 (hidrolizado enzimático de colágeno) y 46 (sulfato de gluco- samina) que, tras una semana de lavado, recibieron o bien 10g de hidrolizado enzimático de colágeno o bien 1.5g de sulfato de glucosamina diarios durante 90 días consecutivos, siendo por Io tanto Ia duración del estudio de 3 meses para cada sujeto. La calidad de vida se midió al inicio y al finalizar ambos tratamientos de modo específico mediante el cuestionario genérico SF-36 validado en multitud de estudios y revisiones bibliográficas y habiendo sido utilizado en más de 4000 artículos científicos a Io largo de los últimos 20 años, siendo originario del Health Institute, New England Medical Center, Boston (Massachussets, USA). En España, dicho cuestio- nario ha sido validado por Alonso y cois. (11) en el IMIM (Instituto Municipal de Investigaciones Médicas) en Barcelona.The subjects were distributed in two groups of 50 individuals, of which 47 (enzymatic collagen hydrolyzate) and 46 (glucoamine sulfate) were evaluated, which, after one week of washing, received either 10g of collagen enzyme hydrolyzate or or 1.5g of glucosamine sulfate daily for 90 consecutive days, being therefore the duration of the study of 3 months for each subject. The quality of life was measured at the beginning and at the end of both treatments in a specific way by means of the generic SF-36 questionnaire validated in many studies and bibliographic reviews and having been used in more than 4000 scientific articles over the last 20 years, originally from the Health Institute, New England Medical Center, Boston (Massachusetts, USA). In Spain, this questionnaire has been validated by Alonso and cois. (11) at the IMIM (Municipal Institute of Medical Research) in Barcelona.
El SF-36 representa para todos los autores que han llevado a cabo diversas validaciones del cuestionario, un instrumento adecuado, veraz y eficaz muy útil en Ia investigación y en Ia práctica clínicas. Con el término "calidad de vida" con el SF-36 se pretendían valorar aspectos de las disfunciones articulares consecutivas a alteraciones de los cartílagos articulares que no son estrictamente clínicos, sino también aspectos relacionados con Ia vida diaria del paciente. También se pretendía valorar en qué modo se veía afectada Ia vida del paciente por Ia presencia de estas disfunciones articulares, siempre desde el punto de vista del propio paciente. Esta valoración fue multidimensional y se aceptó generalmente incluir, al menos, ocho apartados o parámetros que actúan de indicadores de Ia capacidad funcional y/o calidad de vida.The SF-36 represents for all authors who have carried out various validations of the questionnaire, an adequate, truthful and effective instrument very useful in clinical research and practice. With the term "quality of life" with the SF-36, it was intended to assess aspects of joint dysfunctions consecutive to alterations of articular cartilages that are not strictly clinical, but also aspects related to the patient's daily life. It was also intended to assess how the patient's life was affected by the presence of these joint dysfunctions, always from the patient's own point of view. This assessment was multidimensional and it was generally accepted to include at least eight sections or parameters that act as indicators of functional capacity and / or quality of life.
Los parámetros evaluados corresponden a las facetas de Ia vida cotidiana más comúnmente afectadas por las dificultades funcionales derivadas de alteracio- nes de los cartílagos articulares. La división de dichos parámetros nos permite diferenciar con claridad actividades y momentos de Ia vida de las personas en los que el dolor derivado de una alteración articular afecta, o puede afectar, al correcto de desarrollo de actividades y tareas. También se distingue entre Ia afectación de Ia
calidad de vida individual, de Ia relación con otras personas, o en el desempeño de una profesión.The parameters evaluated correspond to the facets of daily life most commonly affected by the functional difficulties derived from alterations of the articular cartilage. The division of these parameters allows us to clearly differentiate activities and moments of the life of people in which the pain derived from an articular alteration affects, or can affect, the correct development of activities and tasks. It also distinguishes between the involvement of Ia quality of individual life, of the relationship with other people, or in the performance of a profession.
Los parámetros evaluados son, y se definen, como sigue.The parameters evaluated are, and are defined, as follows.
1. Función física: Se entiende por función física Ia capacidad para realizar actividades básicas de Ia vida cotidiana como el cuidado personal, Ia movilidad básica y Ia capacidad para realizar esfuerzos pequeños o moderados. Este parámetro nos permite valorar el grado en que Ia alteración de los cartílagos articulares nos limita a título individual.1. Physical function: The physical function is understood as the ability to perform basic activities of daily life such as personal care, basic mobility and the ability to make small or moderate efforts. This parameter allows us to assess the degree to which the alteration of articular cartilage limits us individually.
2. Rol físico: Por su parte, el rol físico se define como el parámetro que nos evalúa Ia capacidad para desempeñar las actividades propias de nuestro puesto de trabajo. Nos permite identificar si nuestra falta de salud afecta a nuestro rendimiento laboral, o nos limita o impide realizar con normalidad las tareas asociadas a nuestro trabajo. Este parámetro nos permite valorar como Ia falta de salud afecta a las actividades físicas relacionadas con nuestro entorno y en el desempeño del trabajo.2. Physical role: For its part, the physical role is defined as the parameter that evaluates the ability to perform the activities of our job. It allows us to identify if our lack of health affects our work performance, or limits us or prevents us from performing normally the tasks associated with our work. This parameter allows us to assess how lack of health affects physical activities related to our environment and work performance.
3. Dolor corporal: Este parámetro nos da Ia medida del dolor percibido y su efecto, tanto en el trabajo y en Ia relación con nuestro entorno, como en las actividades individuales básicas de Ia vida cotidiana.3. Body pain: This parameter gives us the measure of perceived pain and its effect, both at work and in relation to our environment, as well as in the basic individual activities of everyday life.
4. Salud general: Parámetro que valora, por parte del evaluado, Ia percepción del estado de salud que se tiene de uno mismo. Así como las perspectivas futuras y Ia percepción individual de Ia resistencia a enfermar.4. General health: Parameter that assesses, on the part of the evaluated, the perception of one's health status. As well as future perspectives and the individual perception of resistance to illness.
5. Vitalidad: Se define, para Ia interpretación de este estudio, como el sentimiento de energía y vitalidad que se tiene de uno mismo. Engloba también Ia percepción individual de Ia resistencia al cansancio y al desánimo.5. Vitality: It is defined, for the interpretation of this study, as the feeling of energy and vitality that one has of oneself. It also includes the individual perception of resistance to tiredness and discouragement.
6. Función social: Parámetro gracias al cual podemos valorar cómo Ia alte- ración de los cartílagos de las articulaciones, y los problemas derivados de dichas alteraciones, afectan al desarrollo de una vida social satisfactoria. Este parámetro engloba tanto problemas físicos como emocionales. Nos permite valorar como estos problemas interfieren en Ia vida social habitual.6. Social function: Parameter through which we can assess how the alteration of the cartilage of the joints, and the problems derived from these alterations, affect the development of a satisfactory social life. This parameter encompasses both physical and emotional problems. It allows us to assess how these problems interfere with habitual social life.
7. Rol emocional: Se entiende por rol emocional, y así se define para este estudio, el grado en que los problemas emocionales derivados del dolor y Ia afecta-
ción a Ia capacidad física afectan al desempeño del trabajo, así como a Ia realización satisfactoria de otras actividades diarias. La valoración del rol emocional nos da información muy valiosa sobre factores tan importantes como el rendimiento o el esmero en el trabajo.7. Emotional role: It is understood by emotional role, and thus it is defined for this study, the degree to which the emotional problems derived from pain and the affected tion to physical capacity affect work performance, as well as the satisfactory performance of other daily activities. The assessment of the emotional role gives us very valuable information about such important factors as performance or dedication at work.
8. Salud mental: Parámetro que nos permite valorar Ia salud mental general, considerando Ia depresión, Ia ansiedad, el autocontrol y el bienestar general.8. Mental health: Parameter that allows us to assess general mental health, considering depression, anxiety, self-control and general well-being.
A continuación, y a modo de resumen, se describen los ocho apartados o parámetros que actúan de indicadores de Ia capacidad funcional y/o calidad de vida. (Tabla 1)Next, and as a summary, the eight sections or parameters that act as indicators of the functional capacity and / or quality of life are described. (Table 1)
Tabla 1. Parámetros y significado estudiados para determinar Ia calidad de vida de un humano con dificultades funcionales derivadas de alteraciones de los cartílagos articulares.Table 1. Parameters and meaning studied to determine the quality of life of a human with functional difficulties derived from alterations of articular cartilage.
Los resultados obtenidos pueden verse en Ia Tabla 2.
Tabla 2: Resultados del cuestionario genérico SF-36 aplicado a 100 voluntarios afectos de alteraciones articulares que inducían pérdida notable de calidad de vida.The results obtained can be seen in Table 2. Table 2: Results of the generic SF-36 questionnaire applied to 100 volunteers affected by joint disorders that induced a significant loss of quality of life.
Hidrolizado enzimático de colágeno Sulfato de glucosamina m SD me Rango n m SD me Rango nCollagen Enzymatic Hydrolyzate Glucosamine Sulfate m SD me Rank n m SD me Rank n
Visita inicial (día -7)Initial visit (day -7)
Función física 56.0 20.6 55.0 20.0-100.0 47 58.2 21.5 52.5 20.0-100.0 44Physical function 56.0 20.6 55.0 20.0-100.0 47 58.2 21.5 52.5 20.0-100.0 44
Rol físico 36.4 34.8 25.0 0.0-100.0 46 34.8 37.1 25.0 0.0-100.0 46Physical role 36.4 34.8 25.0 0.0-100.0 46 34.8 37.1 25.0 0.0-100.0 46
Dolor corporal 52.5 14.8 52.0 20.0-100.0 46 53.4 14.5 52.0 22.0-84.0 46Body pain 52.5 14.8 52.0 20.0-100.0 46 53.4 14.5 52.0 22.0-84.0 46
Salud General 53.1 16.1 52.0 20.0-97.0 47 54.7 17.1 52.0 25.0-97.0 45General Health 53.1 16.1 52.0 20.0-97.0 47 54.7 17.1 52.0 25.0-97.0 45
Vitalidad 52.9 15.5 52.5 25.0-100.0 46 53.7 14.2 52.5 30.0-100.0 46Vitality 52.9 15.5 52.5 25.0-100.0 46 53.7 14.2 52.5 30.0-100.0 46
Función social 72.3 18.0 75.0 25.0-100.0 46 71.5 20.1 75.0 25.0-100.0 43Social function 72.3 18.0 75.0 25.0-100.0 46 71.5 20.1 75.0 25.0-100.0 43
Rol emocional 82.6 34.2 100.0 0.0-100.0 46 90.6 22.9 100.0 0.0-100.0 46Emotional role 82.6 34.2 100.0 0.0-100.0 46 90.6 22.9 100.0 0.0-100.0 46
Salud mental 66.8 15.5 64.0 28.0-100.0 45 68.9 12.5 68.0 44.0-96.0 44 índice de salud física 36.0 7.7 34.2 23.8-52.8 45 35.8 9.8 34.5 20.4-56.8 39 índice de salud emocional 50.1 9.1 52.1 25.6-66.9 45 51.8 6.8 52.7 29.4-64.9 39Mental health 66.8 15.5 64.0 28.0-100.0 45 68.9 12.5 68.0 44.0-96.0 44 physical health index 36.0 7.7 34.2 23.8-52.8 45 35.8 9.8 34.5 20.4-56.8 39 emotional health index 50.1 9.1 52.1 25.6-66.9 45 51.8 6.8 52.7 29.4- 64.9 39
Visita final (día 90)Final visit (day 90)
Función física 68.22 21.4 70.0 20.0-100.0 47 65.22 23.6 75.0 15.0-100.0 45Physical function 68.2 2 21.4 70.0 20.0-100.0 47 65.2 2 23.6 75.0 15.0-100.0 45
Rol físico 61.22 38.2 75.0 0.0-100.0 47 48.42 42.0 50.0 0.0-100.0 46Physical role 61.2 2 38.2 75.0 0.0-100.0 47 48.4 2 42.0 50.0 0.0-100.0 46
Dolor coφoral 67.11l2 17.0 74.C ) 22.0-100.0 47 62.71'2 16.6 64. 0 22.0-84.0 46Choral pain 67.1 1l2 17.0 74.C) 22.0-100.0 47 62.7 1 ' 2 16.6 64. 0 22.0-84.0 46
Salud General 56.82 13.7 57.0 30.0-97.0 47 57.62 16.0 52.0 25.0-97.0 45General Health 56.8 2 13.7 57.0 30.0-97.0 47 57.6 2 16.0 52.0 25.0-97.0 45
Vitalidad 57.02 16.7 55.0 25.0-100.0 47 57.12 15.7 55.0 30.0-100.0 46Vitality 57.0 2 16.7 55.0 25.0-100.0 47 57.1 2 15.7 55.0 30.0-100.0 46
Función social 81.62 15.2 87.5 50.0-100.0 47 76.12 19.8 75.0 25.0-100.0 44Social function 81.6 2 15.2 87.5 50.0-100.0 47 76.1 2 19.8 75.0 25.0-100.0 44
Rol emocional 89.42 28.7 100.0 0.0-100.0 47 92.82 25.3 100.0 0.0-100.0 46Emotional role 89.4 2 28.7 100.0 0.0-100.0 47 92.8 2 25.3 100.0 0.0-100.0 46
Salud mental 70.32 13.4 68.0 36.0-100.0 47 70.62 13.3 68.0 44.0-96.0 45 índice de salud física 42.02 8.3 44.9 24.8-53.1 47 40.02 9.9 41.0 21.8-56.8 41 índice de salud emocional 50.7 7.7 52.1 25.6-66.5 47 51.1 7.1 52.9 29.6-61.6 41Mental health 70.3 2 13.4 68.0 36.0-100.0 47 70.6 2 13.3 68.0 44.0-96.0 45 physical health index 42.0 2 8.3 44.9 24.8-53.1 47 40.0 2 9.9 41.0 21.8-56.8 41 emotional health index 50.7 7.7 52.1 25.6-66.5 47 51.1 7.1 52.9 29.6-61.6 41
1.-Diferencias estadísticamente significativas entre colatech y glucosamina (GLM, valores repetidos; p<0.05).1.-Statistically significant differences between colatech and glucosamine (GLM, repeated values; p <0.05).
2. -Diferencias estadísticamente significativas entre las visitas inicial y final del mismo tratamiento (GLM, valores repetidos; p<0.05).2.-Statistically significant differences between the initial and final visits of the same treatment (GLM, repeated values; p <0.05).
No se hallaron diferencias estadísticamente significativas entre los valores de ambos grupos de cada visita (GLM, valores repetidos; p<0.05).
De Ia Tabla 2 puede extraerse que tanto el hidrolizado enzimático de colágeno como el sulfato de glucosamlna, administrados durante 90 días consecutivos a dosis de 1Og y de 1.5g /día respectivamente, mejoran notablemente Ia calidad de vida en sus diferentes aspectos. Pero se puede comprobar que inesperadamente los valores de las diferencias absolutas entre Ia visita inicial y final siempre son más amplias con el hidrolizado enzimático de colágeno que con el sulfato de glucosamina.No statistically significant differences were found between the values of both groups of each visit (GLM, repeated values; p <0.05). From Table 2 it can be extracted that both the collagen enzymatic hydrolyzate and glucosamlan sulfate, administered for 90 consecutive days at doses of 1Og and 1.5g / day respectively, significantly improve the quality of life in its different aspects. But it can be verified that unexpectedly the values of the absolute differences between the initial and final visit are always broader with the enzymatic hydrolyzate of collagen than with glucosamine sulfate.
En aspectos tan importantes como Ia salud general, Ia vitalidad, Ia función social, Ia función física, el trabajo físico, Ia función emocional y Ia salud mental po- demos comprobar que el hidrolizado enzimático de colágeno siempre es más eficaz en inducir Ia mejoría de los distintos parámetros investigados que el sulfato de glucosamina.In such important aspects as general health, vitality, social function, physical function, physical work, emotional function and mental health, we can verify that the enzymatic hydrolyzate of collagen is always more effective in inducing the improvement of the different parameters investigated than glucosamine sulfate.
Del análisis de los resultados globales obtenidos en este estudio clínico, se ponen de manifiesto las ventajas de utilizar el hidrolizado enzimático de colágeno objeto de Ia invención para Ia preparación de un alimento funcional o suplemento nutricional para mejorar Ia capacidad funcional y/o Ia calidad de vida de las personas con dificultades funcionales derivadas de alteraciones de los cartílagos articulares.From the analysis of the overall results obtained in this clinical study, the advantages of using the enzymatic collagen hydrolyzate object of the invention for the preparation of a functional food or nutritional supplement to improve the functional capacity and / or the quality of life of people with functional difficulties derived from alterations of articular cartilage.
Con el hidrolizado enzimático de colágeno de alta pureza objeto de Ia inven- ción dentro de las dosis por Kg de peso corporal empleadas en el estudio clínico (0,1 - 0,25 g/Kg peso corporal/día) pueden pues preparase debido a su buen sabor y sus buenas características organolépticas, distintos tipos de complementos o suplementos nutricionales como soluciones o suspensiones bebibles, mezclas de polvos dispersables en agua, comprimidos, cápsulas. Del mismo modo pueden prepa- rarse alimentos funcionales tales como zumos, yogures, yogures líquidos, natillas, barritas energéticas, barritas adelgazantes, etc., a Ia vez que el mismo hidrolizado enzimático de colágeno por su buen sabor puede ser en sí mismo el propio alimento funcional.With the enzymatic hydrolyzate of high purity collagen object of the invention within the doses per Kg of body weight used in the clinical study (0.1 - 0.25 g / Kg body weight / day) can therefore be prepared due to its good taste and good organoleptic characteristics, different types of nutritional supplements or supplements such as drinkable solutions or suspensions, mixtures of water dispersible powders, tablets, capsules. In the same way, functional foods such as juices, yogurts, liquid yogurts, custards, energy bars, slimming bars, etc. can be prepared, at the same time that the same collagen enzymatic hydrolyzate for its good taste can itself be itself functional food.
Como conclusión, queda firmemente demostrado que además el procedi- miento de obtención de un hidrolizado de colágeno objeto de Ia invención, que valoriza los subproductos residuales contaminantes procedentes de Ia obtención de gelatinas, no únicamente permite rentabilizar los costes derivados del tratamiento de estos subproductos nocivos por el medio ambiente, sino que también permite obtener un nuevo hidrolizado enzimático de colágeno de alta pureza y bajo peso
molecular que debido a su buen sabor y características organolépticas puede emplearse tanto en alimentos funcionales, tales como yogures, zumos, postres, barritas energéticas, etc., como en suplementos nutricionales del tipo complejos multivi- tamínicos en forma de comprimidos, soluciones, suspensiones, etc. Además, los inventores han descubierto un nuevo uso para este hidrolizado enzimático de colágeno, que permite que Ia calidad de vida de las personas o animales afectos de disfunciones derivadas de alteraciones en los cartílagos de las articulaciones, pueda verse notablemente mejorada de una manera equivalente y más agradable a Ia administración de antiinflamatorios, eliminándose los indesea- dos efectos secundarios de estos últimos.
In conclusion, it is clearly demonstrated that in addition the procedure for obtaining a collagen hydrolyzate object of the invention, which valorizes the residual contaminant by-products derived from the obtaining of jellies, not only allows the costs derived from the treatment of these harmful by-products to be profitable for the environment, but also allows to obtain a new enzymatic hydrolyzate of collagen of high purity and low weight molecular that due to its good taste and organoleptic characteristics can be used both in functional foods, such as yogurts, juices, desserts, energy bars, etc., and in nutritional supplements such as multivitamin complexes in the form of tablets, solutions, suspensions, etc. In addition, the inventors have discovered a new use for this collagen enzymatic hydrolyzate, which allows the quality of life of people or animals affected by dysfunctions derived from alterations in the cartilage of the joints, can be markedly improved in an equivalent manner and more pleasant to the administration of anti-inflammatories, eliminating the undesirable side effects of the latter.
Claims
1.- Hidrolizado enzimático de colágeno con un peso molecular comprendido entre 1.000 y 50.000 Daltons, caracterizado porque tiene una acidez total, expresa- da en grados Dornic, inferior a 250D; y una conductividad en solución al 1 % en peso no superior a 500 μS/cm.1.- Enzymatic hydrolyzate of collagen with a molecular weight between 1,000 and 50,000 Daltons, characterized in that it has a total acidity, expressed in Dornic degrees, less than 25 0 D; and a conductivity in 1% by weight solution not exceeding 500 μS/cm.
2.- Hidrolizado enzimático de colágeno según Ia reivindicación 1 , caracterizado porque Ia acidez total, expresada en grados Dornic, es inferior a 250D, preferi- blemente inferior o igual a 2O0D.2.- Enzymatic hydrolyzate of collagen according to claim 1, characterized in that the total acidity, expressed in Dornic degrees, is less than 25 0 D, preferably less than or equal to 2O 0 D.
3.- Hidrolizado enzimático de colágeno según una cualquiera de las reivindicaciones anteriores, caracterizado porque tiene un peso molecular comprendido entre 5.000 y 20.000 Daltons.3.- Enzymatic hydrolyzate of collagen according to any one of the preceding claims, characterized in that it has a molecular weight between 5,000 and 20,000 Daltons.
4.- Hidrolizado enzimático de colágeno según una cualquiera de las reivindicaciones anteriores, caracterizado porque comprende un 99% en peso de hidrolizado sobre producto seco.4.- Enzymatic hydrolyzate of collagen according to any one of the preceding claims, characterized in that it comprises 99% by weight of hydrolyzate based on dry product.
5.- Procedimiento de obtención de un hidrolizado de colágeno de baja acidez total a partir de tejido dérmico de origen animal, en su forma nativa, o a partir de subproductos de procesos de obtención de gelatina, en los que después de Ia extracción en medio acuoso de las gelatinas, los tejidos descritos, ricos en colágeno, son sometidos a sucesivas etapas de extracción acuosa y posterior hidrólisis acida, básica o enzimática, obteniéndose finalmente un subproducto residual, caracterizado porque comprende las etapas de: a) extracción acuosa del subproducto residual a una temperatura comprendida entre 4O0C y 1400C durante un tiempo comprendido entre 2 y 8 horas; b) ajuste del pH entre 6,5 y 8,0 mediante soluciones de ácidos o bases mi- nerales; c) filtración de Ia solución obtenida en Ia etapa b); d) desmineralización de Ia solución filtrada por paso de Ia misma a través de resinas de intercambio iónico y/o equipos de diálisis por membranas hasta un régimen estacionario en el que Ia conductividad absoluta de Ia solución no supere los
5.- Procedure for obtaining a collagen hydrolyzate of low total acidity from dermal tissue of animal origin, in its native form, or from by-products of gelatin production processes, in which after extraction in an aqueous medium of the gelatins, the tissues described, rich in collagen, are subjected to successive stages of aqueous extraction and subsequent acid, basic or enzymatic hydrolysis, finally obtaining a residual by-product, characterized in that it comprises the steps of: a) aqueous extraction of the residual by-product a temperature between 40 0 C and 140 0 C for a time between 2 and 8 hours; b) adjustment of the pH between 6.5 and 8.0 using solutions of mineral acids or bases; c) filtration of the solution obtained in stage b); d) demineralization of the filtered solution by passing it through ion exchange resins and/or membrane dialysis equipment to a steady state in which the absolute conductivity of the solution does not exceed the
1.000 μS/cm; e) ultrafiltración de Ia solución desmineralizada a través de membranas con un diámetro de poro de 1.000 a 40.000 Daltons; f) hidrólisis enzimática de Ia solución ultrafiltrada con una cantidad de enzi- ma comprendida entre 1.000 Ul y 4.000 Ul por kilogramo de producto seco a hidro- lizar y a un pH comprendido entre el pH óptimo de Ia enzima ± 0,5 unidades de pH ; g) filtración de Ia solución de colágeno hidrolizado enzimáticamente; h) ultrafiltración a través de membranas con un diámetro de poro de 10.000 a 40.000 Daltons; i) concentración de Ia solución de hidrolizado enzimático de colágeno mediante calefacción a temperaturas inferiores o iguales a 7O0C a una presión comprendida entre 10 y 0,0001 mm de Hg hasta una concentración final de hidrolizado enzimático de colágeno del 15% al 45% en peso; j) secado del hidrolizado enzimático de colágeno hasta obtener un producto seco con una humedad inferior o igual al 8% y una densidad entre 0,3 y 0,8 g/ml.1,000 μS/cm; e) ultrafiltration of the demineralized solution through membranes with a pore diameter of 1,000 to 40,000 Daltons; f) enzymatic hydrolysis of the ultrafiltered solution with an amount of enzyme between 1,000 Ul and 4,000 Ul per kilogram of dry product to be hydrolyzed and at a pH between the optimal pH of the enzyme ± 0.5 pH units; g) filtration of the enzymatically hydrolyzed collagen solution; h) ultrafiltration through membranes with a pore diameter of 10,000 to 40,000 Daltons; i) concentration of the enzymatic collagen hydrolyzate solution by heating at temperatures lower than or equal to 7O 0 C at a pressure between 10 and 0.0001 mm Hg up to a final concentration of collagen enzymatic hydrolyzate of 15% to 45% in weigh; j) drying the enzymatic collagen hydrolyzate until obtaining a dry product with a humidity less than or equal to 8% and a density between 0.3 and 0.8 g/ml.
6.- Procedimiento según Ia reivindicación 5, caracterizado porque en Ia etapa f) se realiza Ia hidrólisis enzimática de Ia solución ultrafiltrada con una cantidad de enzima comprendida entre 1.500 Ul y 2.500 Ul por kilogramo de producto seco a hidrolizar y a un pH comprendido entre el pH óptimo de Ia enzima ± 0,5 unidades de pH.6.- Procedure according to claim 5, characterized in that in step f) the enzymatic hydrolysis of the ultrafiltered solution is carried out with an amount of enzyme between 1,500 Ul and 2,500 Ul per kilogram of dry product to be hydrolyzed and at a pH between Optimum pH of the enzyme ± 0.5 pH units.
7.- Procedimiento de obtención de hidrolizado de colágeno según una cualquiera de las reivindicaciones 5 ó 6 anteriores, caracterizado porque Ia etapa a) de extracción acuosa del subproducto residual se realiza a una temperatura comprendida entre 450C y 1000C, concretamente entre 5O0C y 8O0C durante un tiempo comprendido entre 4 y 6 horas.7.- Procedure for obtaining collagen hydrolyzate according to any one of the previous claims 5 or 6, characterized in that step a) of aqueous extraction of the residual byproduct is carried out at a temperature between 45 0 C and 100 0 C, specifically between 5O 0 C and 8O 0 C for a time between 4 and 6 hours.
8.- Procedimiento de obtención de hidrolizado de colágeno según una cual- quiera de las reivindicaciones 5 a 7 anteriores, caracterizado porque después de Ia extracción acuosa se ajusta el pH de Ia solución entre 7,4 y 7,6.8.- Procedure for obtaining collagen hydrolyzate according to any of the preceding claims 5 to 7, characterized in that after the aqueous extraction the pH of the solution is adjusted between 7.4 and 7.6.
9.- Procedimiento de obtención de hidrolizado de colágeno según una cualquiera de las reivindicaciones 5 a 8 anteriores, caracterizado porque Ia etapa de
ultrafiltración de Ia solución de colágeno hidrolizado enzimáticamente, se lleva a cabo a través de membranas con un diámetro de poro de 5.000 a 20.000 Daltons.9.- Procedure for obtaining collagen hydrolyzate according to any one of claims 5 to 8 above, characterized in that the stage of Ultrafiltration of the enzymatically hydrolyzed collagen solution is carried out through membranes with a pore diameter of 5,000 to 20,000 Daltons.
10.- Procedimiento de obtención de hidrolizado de colágeno según una cual- quiera de las reivindicaciones 5 a 9 anteriores, caracterizado porque Ia etapa de concentración i) de Ia solución de hidrolizado enzimático de colágeno se realiza a una temperatura inferior o igual a 6O0C.10.- Procedure for obtaining collagen hydrolyzate according to any of the preceding claims 5 to 9, characterized in that the concentration step i) of the enzymatic collagen hydrolyzate solution is carried out at a temperature lower than or equal to 6O 0 c.
11.- Procedimiento de obtención de hidrolizado de colágeno una cualquiera de las reivindicaciones 5 a 10 anteriores, caracterizado porque Ia etapa de concentración i) de Ia solución de hidrolizado enzimático de colágeno se realiza a una presión comprendida entre 1,0 y 0,01 mm de Hg.11.- Procedure for obtaining collagen hydrolyzate from any of the preceding claims 5 to 10, characterized in that the concentration step i) of the enzymatic collagen hydrolyzate solution is carried out at a pressure between 1.0 and 0.01. mm of Hg.
12.-Alimento funcional que comprende el hidrolizado enzimático de colágeno según una cualquiera de las reivindicaciones 1 a 4 anteriores.12.-Functional food comprising the enzymatic hydrolyzate of collagen according to any one of claims 1 to 4 above.
13.- Alimento funcional según Ia reivindicación 12, caracterizado porque comprende leche cruda o tratada térmicamente.13.- Functional food according to claim 12, characterized in that it comprises raw or heat-treated milk.
14.- Alimento funcional según las reivindicaciones 12 ó 13 que además comprende un compuesto seleccionado independientemente de entre: compuestos antioxidantes, sales minerales, vitamina y/o coenzimas, extractos vegetales, extractos de origen animal y compuestos coadyuvantes del hidrolizado enzimático de colágeno, o una mezcla de todos ellos.14.- Functional food according to claims 12 or 13 that also comprises a compound independently selected from: antioxidant compounds, mineral salts, vitamins and/or coenzymes, plant extracts, extracts of animal origin and adjuvant compounds of the enzymatic hydrolyzate of collagen, or a mix of all of them.
15.- Suplemento nutricional a base de leche que comprende el hidrolizado enzimático de colágeno según una cualquiera de las reivindicaciones 1 a 4 anteriores.15.- Milk-based nutritional supplement comprising the enzymatic hydrolyzate of collagen according to any one of claims 1 to 4 above.
16.- Suplemento nutricional según Ia reivindicación 15 que además comprende un compuesto seleccionado independientemente de entre: compuestos antioxidantes, sales minerales, vitamina y/o coenzimas, extractos vegetales, extractos de origen animal y compuestos coadyuvantes del hidrolizado enzimático de colágeno, o una mezcla de todos ellos.
16.- Nutritional supplement according to claim 15 that also comprises a compound independently selected from: antioxidant compounds, mineral salts, vitamins and/or coenzymes, plant extracts, extracts of animal origin and adjuvant compounds of the enzymatic hydrolyzate of collagen, or a mixture of all of them.
17.- Uso del hidrolizado enzimático de colágeno según una cualquiera de las reivindicaciones 1 a 4 para Ia preparación de un producto lácteo, suplemento nutri- cional o alimento funcional para mejorar Ia capacidad funcional y/o Ia calidad de vida de un mamífero con dificultades funcionales derivadas de alteraciones de los cartílagos articulares.17.- Use of the enzymatic collagen hydrolyzate according to any one of claims 1 to 4 for the preparation of a dairy product, nutritional supplement or functional food to improve the functional capacity and/or the quality of life of a mammal with difficulties functional derived from alterations of the articular cartilage.
18.- Uso según Ia reivindicación 17 para Ia preparación de un producto lácteo, suplemento nutricional o alimento funcional para mejorar Ia función física.18.- Use according to claim 17 for the preparation of a dairy product, nutritional supplement or functional food to improve physical function.
19.- Uso según Ia reivindicación 17 para Ia preparación de un producto lácteo, suplemento nutricional o alimento funcional para Ia mejora del rol físico.19.- Use according to claim 17 for the preparation of a dairy product, nutritional supplement or functional food for the improvement of the physical role.
20.- Uso según Ia reivindicación 17 para Ia preparación de un producto lácteo, un suplemento nutricional o alimento funcional para mejorar Ia sensación de dolor corporal.20.- Use according to claim 17 for the preparation of a dairy product, a nutritional supplement or functional food to improve the sensation of body pain.
21.- Uso según Ia reivindicación 17 para Ia preparación de un producto lácteo, un suplemento nutricional o alimento funcional para mejorar Ia salud general.21.- Use according to claim 17 for the preparation of a dairy product, a nutritional supplement or functional food to improve general health.
22.- Uso según Ia reivindicación 17 para Ia preparación de un producto lácteo, un suplemento nutricional o alimento funcional para mejorar Ia vitalidad.22.- Use according to claim 17 for the preparation of a dairy product, a nutritional supplement or functional food to improve vitality.
23.- Uso según Ia reivindicación 17 para Ia preparación de un producto lácteo, un suplemento nutricional o alimento funcional para mejorar Ia función social.23.- Use according to claim 17 for the preparation of a dairy product, a nutritional supplement or functional food to improve social function.
24.- Uso según Ia reivindicación 17 para Ia preparación de un producto lácteo, un suplemento nutricional o alimento funcional para mejorar del rol emocional.24.- Use according to claim 17 for the preparation of a dairy product, a nutritional supplement or functional food to improve the emotional role.
25.- Uso según Ia reivindicación 17 para Ia preparación de un producto lác- teo, un suplemento nutricional o alimento funcional para mejorar Ia salud mental.25.- Use according to claim 17 for the preparation of a dairy product, a nutritional supplement or functional food to improve mental health.
26.- Uso según una cualquiera de las reivindicaciones 17 a 25 anteriores, donde Ia dosis diaria de hidrolizado enzimático de colágeno está comprendida entre 0,1 y 0,25 g/Kg de peso corporal.
26.- Use according to any one of the preceding claims 17 to 25, wherein the daily dose of collagen enzymatic hydrolyzate is between 0.1 and 0.25 g/kg of body weight.
27.- Uso según una cualquiera de las reivindicaciones 17 a 25 anteriores, caracterizado porque es para una preparación en forma de soluciones o suspensiones orales.27.- Use according to any one of the preceding claims 17 to 25, characterized in that it is for a preparation in the form of oral solutions or suspensions.
28.- Uso según una cualquiera de las reivindicaciones 17 a 25 anteriores, caracterizado porque es para una preparación en forma de comprimidos, cápsulas o polvos dispersables en agua.28.- Use according to any one of claims 17 to 25 above, characterized in that it is for a preparation in the form of tablets, capsules or powders dispersible in water.
29.- Uso según una cualquiera de las reivindicaciones 17 a 28 anteriores, en donde el mamífero con dificultades funcionales derivadas de alteraciones de los cartílagos articulares es un humano.
29.- Use according to any one of claims 17 to 28 above, wherein the mammal with functional difficulties derived from alterations of the articular cartilage is a human.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ESESP200602713 | 2006-10-25 | ||
| ES200602713A ES2298071B1 (en) | 2006-10-25 | 2006-10-25 | HYDROLYZED ENZYMATIC COLLAGEN AND PROCEDURE OF OBTAINING. |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2008049942A1 true WO2008049942A1 (en) | 2008-05-02 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/ES2007/000593 WO2008049942A1 (en) | 2006-10-25 | 2007-10-22 | Enzymatic collagen hydrolysate and method for obtaining same |
Country Status (2)
| Country | Link |
|---|---|
| ES (1) | ES2298071B1 (en) |
| WO (1) | WO2008049942A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10226422B2 (en) | 2013-01-23 | 2019-03-12 | Bottled Science Limited | Skin enhancing beverage composition |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3967151A1 (en) * | 2020-07-14 | 2022-03-16 | Goia Limited | Valorisation of marine animal by-products |
| WO2023111607A1 (en) * | 2021-12-15 | 2023-06-22 | Goia Limited | Valorisation of marine animal by-products |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2212649A1 (en) * | 1997-08-08 | 1999-02-08 | Biocell Technology, Llc | Hydrolyzed collagen type ii and use thereof |
| US20030175398A1 (en) * | 2000-05-09 | 2003-09-18 | Nobuhiro Ogasawara | Stable acidic milk drinks, process for producing the same, and additive for acidic milk drinks to be used therein |
| WO2005021027A1 (en) * | 2003-08-21 | 2005-03-10 | Gelita Ag | Collagen hydrolysate |
| ES2233947T3 (en) * | 1994-08-23 | 2005-06-16 | Deutsche Gelatine-Fabriken Stoess Ag | EMPLOYMENT OF HYDROLYZED COLLAGEN, NEUTRAL FLAVOR, AND AGENT CONTAINING IT. |
| WO2006010606A1 (en) * | 2004-07-28 | 2006-02-02 | Cargill, Incorporated | Physiologically-active composition based on collagen |
-
2006
- 2006-10-25 ES ES200602713A patent/ES2298071B1/en not_active Expired - Fee Related
-
2007
- 2007-10-22 WO PCT/ES2007/000593 patent/WO2008049942A1/en active Application Filing
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2233947T3 (en) * | 1994-08-23 | 2005-06-16 | Deutsche Gelatine-Fabriken Stoess Ag | EMPLOYMENT OF HYDROLYZED COLLAGEN, NEUTRAL FLAVOR, AND AGENT CONTAINING IT. |
| CA2212649A1 (en) * | 1997-08-08 | 1999-02-08 | Biocell Technology, Llc | Hydrolyzed collagen type ii and use thereof |
| US20030175398A1 (en) * | 2000-05-09 | 2003-09-18 | Nobuhiro Ogasawara | Stable acidic milk drinks, process for producing the same, and additive for acidic milk drinks to be used therein |
| WO2005021027A1 (en) * | 2003-08-21 | 2005-03-10 | Gelita Ag | Collagen hydrolysate |
| WO2006010606A1 (en) * | 2004-07-28 | 2006-02-02 | Cargill, Incorporated | Physiologically-active composition based on collagen |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10226422B2 (en) | 2013-01-23 | 2019-03-12 | Bottled Science Limited | Skin enhancing beverage composition |
Also Published As
| Publication number | Publication date |
|---|---|
| ES2298071A1 (en) | 2008-05-01 |
| ES2298071B1 (en) | 2009-07-28 |
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