RU2743860C1 - Method for obtaining collagen extract from broiler chicken legs - Google Patents

Abstract

FIELD: collagen extract production.SUBSTANCE: invention relates to method for producing collagen extract from the legs of broiler chickens. The method provides for the purification of raw materials, washing, grinding, hydrolysis and extraction with acetic acid solution, centrifugation. Hydrolysis is carried out with a 2.0-2.5% solution of acetic acid with exposure for 6-10 hours at 18-25°C, and the extraction of raw materials is carried out with a 1.5-1.8% solution of acetic acid at temperature 50-55°С for 45-50 minutes with constant stirring.EFFECT: provides high yield of collagen extract from broiler chickens legs and shortens the production cycle.3 cl, 4 ex

Description

Technology area

The invention relates to the food, medical and cosmetic industry, in particular to a method for producing collagen extract, which can find its application in the food industry for the production of food products as a structure former in the production of canned products, as a basis for the production of food and edible coatings for products, broths, jellies, sauces, drinks, additives in the bakery and confectionery industries, for use as an additive that allows the texturing of food products, as well as in medicine, as a remedy for the treatment of damaged tissues, and in the cosmetic industry - in the production of cosmetics.

State of the art

Collagen is a widespread group of proteins found only in the animal kingdom. In collagen, there are several main sequential structural levels of its structure. Collagen structure is divided into primary, secondary, tertiary and quaternary. The structure of collagen isolated from the dermis, tendons, cartilage, bone tissue and other parts of the body is identical at the primary, secondary and tertiary structural levels. The main difference between these types of collagen can be found mainly at the Quaternary and submicroscopic levels. The quaternary and microstructure is influenced not only by the type of organism or the nature of its tissue containing collagen, but also by a number of other factors, for example, age. The structure of collagen contains atoms of nitrogen, carbon, oxygen, hydrogen and sulfur. Collagen differs from most other proteins by its increased nitrogen content, on average 17.8%. Therefore, the conversion factor of nitrogen to protein for collagen is 5.62 (100: 17.8 = 5.62).

In terms of amino acid composition, collagen occupies a special place among proteins. It is the only protein of animal origin that contains large amounts of glycine, hydroxyproline and proline, and small amounts of tyrosine and methionine. Collagen contains very little sulfur, and methionine is the only amino acid that contains sulfur. Cysteine, cystine and tryptophan are absent. These features of the composition of collagen determine its properties. The complexity of the collagen structure determines the variety of changes that are observed during certain treatments. One of the most characteristic properties of collagen is its ability to convert into a soluble gelling agent, which can be obtained by heating in hot water. If collagen is subjected to extraction in hot water in a neutral medium without additional processing, then the amount of dissolved protein is insignificant. To obtain the maximum yield of collagen with good physicochemical properties, it is necessary to prepare collagen-containing raw materials, i.e. pretreat. Collagen preparation involves breaking it down. The breakdown of collagen is understood as a set of processes for dividing its structure into elements of a lower order with the destruction of various bonds that can be caused by the action of acids, alkalis, salts, enzymes, etc. The nature of the resulting products depends on the method of preparation of the raw materials (alkaline, acidic, etc.). When preparing collagen, it is necessary to achieve such a degree of destruction of strong intermolecular bonds in its structure so that the melting of the product proceeds at a relatively low temperature (50-60 ° C) and that for a short time of heat exposure the product yield is maximum (at least 70 wt%).

The older the animal, the more strong cross-linking of the covalent type in collagen and the more difficult it is to dissolve during various treatments.

The use of drugs and materials from collagen in medicine, especially in reconstructive surgery, has been widely covered. The main advantage of collagen as a plastic material is the absence of toxic and carcinogenic properties, high mechanical strength and resistance to tissue enzymes, as well as stimulation of the regeneration of the body's own tissues. In this respect, collagen preparations have advantages over preparations from other biopolymers, in particular from fibrin or gelatin, which is denatured collagen.

Usually, protein preparations are of limited use for healing wounds, burns, stimulating skin metabolism and regeneration, as they often cause allergies. Hydrolysates from tissues of young animals and birds, taken before their immunological maturation, do not possess allergenicity, in addition, embryonic tissues have the ability to stimulate and normalize the exchange of damaged and age-related tissue changes. For example, the drug "Solcoseryl" of the Swiss company "Salk", which is widely used not only for the treatment of wounds and burns, but also for the treatment of stomach and duodenal ulcers, trophic ulcers, etc. The principle of creation is the same - embryonic-regulatory properties of immunologically immature tissues (the drug "Solcoseryl" is obtained from the blood of calves).

From the standpoint of the use of collagen extract in food, along with the indicators necessary in this case, it was found that it has a high digestibility (89.3%), the ability to gel at room temperature, even with threefold dilution, and maintain the shape of the product for a long time. Biological activity confirmed by research, rather high content of such important microelements as chromium, calcium, zinc, silicon involved in the regulation of carbohydrate metabolism, good emulsifying and gel-forming ability is the basis for the use of collagen extract as a structure-forming agent in the development of food products based on it. On the basis of clinical, sanitary-chemical and sanitary-microbiological tests, the developed products (mousses, jellies, soufflés) are recommended for the preventive nutrition of people and patients with diseases of the digestive system and suffering from various metabolic diseases (obesity, diabetes mellitus, hypertension).

From the prior art, various methods are known for the production of collagen extract from the legs of freshly slaughtered broiler chickens at the age of 7-8 weeks with their immunological immaturity, by extraction with a solution of acetic acid. The resulting collagen extract is an aqueous solution of peptides with different molecular weights and amino acids that have undergone a minimum of biological degradation and contains dissolved trace elements.

A known method of producing collagen by grinding biological material (cattle dermis), processing with a saturated solution of sodium sulfate containing 10% sodium hydroxide solution, with an exposure time of 3 to 5 days, neutralizing alkali with an aqueous solution of boric acid, washing with running water, dissolving the treated material in one-molar acetic acid and purification by dialysis against distilled water for 3 days (Khilkin AM, Shekhter A.B., Istranov L.P., Lemenev V.L. Collagen and its use in medicine, 1976. - M. Medicine. - p. . 35).

The disadvantages of this method are the long period of time required to isolate the finished product, the use of expensive reagents used to obtain collagen and environmental pollution with wastewater containing inorganic acids and alkalis.

There is also a known method of obtaining collagen from the legs of broiler chickens, which involves grinding the legs of broiler chickens, mixing the crushed raw material with a 5% solution of acetic acid, taken per 1 part of the raw material, use 8 parts of the acid solution, keeping the resulting mass at a temperature of 4-7 ° C in for 12, 24, 36 and 48 hours and centrifugation. (Optimum Condition of Extracting Collagen from Chiken Feet and its Characteristics / Asian Australisian Journal of Animal Sciences, November 2001, 14 (11): 1638-1644). The collagen yield according to this method is 17-19% of the feedstock.

The disadvantage of this method is the low yield of collagen, the duration of the technological process (from 3 to 5 days), as well as an excess of acetic acid.

The closest in technical essence and the achieved result to the proposed invention is a method of obtaining a protein hydrolyzate from the legs of broiler chickens for medical use, which consists in cleaning raw materials, grinding the purified raw materials and keeping them in dilute acid, the resulting mixture is decanted, filtered or filtered, and then dehydrated or concentrated to a dry powder or dried gel (US 4455302, IPC A61K 35/12). The crushed raw materials are mixed with 1.0-1.5% acetic acid solution and kept at room temperature for 8 to 12 hours with periodic stirring. Then the crushed mixture is washed with water and re-poured with a solution of acetic acid with pH = 3.6-6.5, preferably 4.6, at a rate of 1: 1 by weight and the resulting mixture is heated to a temperature of 54-60 ° C and kept at this temperature within 30-40 minutes with constant stirring. The resulting mixture is centrifuged to remove fat and particulate matter and dried if necessary.

The disadvantages of this method are that the low concentration of acetic acid does not contribute to an increase in the technological process, and the temperature mode of extraction leads to the conversion of collagen into gelatin.

Disclosure of invention

The objective of the present invention is to develop a simple effective method for obtaining collagen extract from the legs of broiler chickens using a waste-free technology.

The solution to the technical problem is achieved by the fact that the method of obtaining collagen extract from the legs of broiler chickens, which provides for the stages of raw material purification, washing, grinding, hydrolysis and extraction with acetic acid solution, and centrifugation, while hydrolysis is carried out 2.0-2.5% - a solution of acetic acid with exposure for 6-10 hours at a temperature of 18-25 ° C, and the extraction of raw materials is carried out with a 1.5-1.8% solution of acetic acid at a temperature of 50-55 ° C for 45-50 minutes with constant stirring.

The technical result of the claimed invention is to increase the yield of collagen to at least 70 mass. % and reduction of the technological cycle (from 10 to 15 hours).

Implementation of the invention

Currently, broiler chickens enter processing at the age of 5-6 weeks (35-42 days) due to the fact that newly hatched crosses reach a carcass weight of 1.5-1.8 kg at this age. In this regard, the resulting slaughter products, such as legs, are immunologically immature raw materials, i.e. are devoid of antigenicity and the resulting collagen protein does not cause allergies.

The raw materials are sorted out from mamins and calcareous growths, freed from the remnants of the keratinized scaly layer of the epidermis, the remnants of scales, feathers, and impurities are removed. Rinse with tap water. The cleaned and washed legs are crushed, grinding can be carried out on an electric meat grinder or a top with a grid hole diameter of 5-6 mm. The resulting crushed mass is weighed. Then, a 2.0-2.5% aqueous solution of glacial acetic acid was prepared in the reactor and crushed legs are loaded into it in a ratio of 1: 1.2 - 1.2: 1 by weight, preferably 1: 1. The mixture is left for 6-10 hours at a temperature of 18-25 ° for hydrolysis, stirring occasionally to ensure maximum contact of the raw material with the acetic acid solution.

To remove hydrolysis products, blood, most of the fat and other impurities, the raw materials are washed with tap water. It is possible and preferable to wash on a mesh with aperture size of 2-3 mm.

Separation of raw materials from wash water is carried out on a batch-type settling centrifuge. The raw material, finally washed and separated from water, is weighed. For the subsequent extraction of collagen protein from the prepared raw material, a 1.5-1.8% aqueous solution of glacial acetic acid was prepared in a ratio of 1: 1.2 - 1.2: 1, preferably 1: 1, to the mass of the raw material with the pH of the mixture should be in the range of 4.0-4.5. The extraction of raw materials is carried out in tilting boilers with mechanical stirrers, maintaining the temperature of the mixture at 50-55 ° C for 45-50 minutes. with constant stirring. The resulting extract is fed into a collection tank equipped with a heating jacket, and the meat and bone meal is unloaded into containers and used for fodder purposes as a mineral supplement. Next, the extract is purified by filtration on a Druk filter. Preliminarily, a precoated layer of filtered perlite of group B with a thickness of 25-30 mm is applied to the filter fabric. Filtration is carried out by creating pressure with compressed air in the collector, from which the extract flows to the Druk filter to ensure a uniform and gradual rate of passage of the extract. The purified extract is collected in a collection tank equipped with a heating jacket. During filtration, the temperature of the extract is maintained within the range of 50-55 ° C. The yield of the extract in a gel state is 78-80% of the mass of the prepared raw material (cleaned, washed and crushed legs of broiler chickens). At a temperature of the collagen extract from 50 ° C to 37 ° C, it is a colorless, sticky, slightly opalescent liquid, and at room temperature (18-25 ° C), the extract is an elastic, transparent gel that persists for a long time without syneresis.

Using in the method as raw material the legs of 5-6 week old broiler chickens and carrying out the extraction at a temperature of 50-55 ° C for 45-50 minutes using a low concentration of acetic acid allows you to obtain collagen with a yield of at least 70 wt. %.

The invention is illustrated by examples.

Example 1. The raw material, namely the legs of broiler chickens, separated from the carcasses of broiler chickens 5-6 weeks of age, were sorted from mamin and calcareous growths, freed from the remnants of the keratinized scaly layer of the epidermis, removed the remnants of scales, feathers, dirt. Washed with tap water in perforated drums. The cleaned and washed legs were ground in an electric meat grinder. The resulting milled mass was weighed. Then, a 2% aqueous solution of glacial acetic acid was prepared in the reactor and crushed legs are loaded into it in a ratio of 1.2: 1 by weight. The mixture was left for 6 h at a temperature of 18 ° C for hydrolysis, stirring occasionally to ensure maximum contact of the raw material with the acetic acid solution.

To remove hydrolysis products, blood, most of the fat and other impurities, the raw materials were washed twice with tap water on a mesh with a mesh size of 2 mm. Separation of raw materials from wash water was carried out on a batch-type settling centrifuge.

The raw material, finally washed and separated from water, was weighed. For the subsequent extraction of collagen protein from the prepared raw material, a 1.5% aqueous solution of glacial acetic acid was prepared in a ratio of 1.2: 1 to the weight of the raw material, while the pH of the mixture was 4.1. The extraction of raw materials is carried out in tilting boilers with mechanical stirrers, maintaining the temperature of the mixture at 55 ° C for 45 minutes. The resulting extract was fed into a collection tank equipped with a heating jacket, and meat and bone meal was unloaded into containers and used for fodder purposes as a mineral supplement. Next, the extract was purified by filtration on a Druk filter. Preliminarily, on the filter cloth, a precoated layer of filtered perlite of group B with a thickness of 25 mm was applied. Filtration was carried out by creating pressure with compressed air in the collector, from which the extract flows to the Druk filter. The purified extract was collected in a collection tank equipped with a heating jacket. During filtration, the temperature of the extract was maintained at 55 ° C. The yield of the extract in the gel state was 78% of the mass of the prepared raw material (cleaned, washed and crushed legs of broiler chickens). At a temperature of the collagen extract from 50 ° C to 37 ° C, it is a colorless, sticky, slightly opalescent liquid, and at room temperature (18-25 ° C), the extract is an elastic, transparent gel that persists for a long time (more than 1 year) without syneresis phenomena. The time taken to obtain the extract was 10 hours.

When alkali was added to a liquid solution of collagen, it became cloudy and precipitated in the form of fibrous structures with properties characteristic of native (natural) collagen ("Collagen and its application in medicine" Khilkin AM, Shekhter A.B. et al., 1976, p. . 33-35).

The remaining meat-and-bone meal, according to the conclusion of the cynologists, can be used as a mineral additive in animal feed, because contains a fairly large amount of calcium, magnesium and potassium (in mg / 100 g - 2146; 26.2; 10.8 - respectively). Thus, the technology for producing collagen extract is waste-free and environmentally friendly, because for its production, food-grade acetic acid of low (1.5-1.8%) concentration is used.

Example 2.

The initial raw materials, namely the legs of broiler chickens, separated from the carcasses of broiler chickens of 5-6 weeks of age, were sorted from mamin and calcareous outgrowths, freed from the remnants of the keratinized scaly layer of the epidermis, removed the remnants of scales, feathers, dirt. Washed with tap water in perforated drums. The cleaned and washed legs were grinded on a top with a grid hole diameter of 5 mm. The resulting milled mass was weighed. Then, a 2.5% aqueous solution of glacial acetic acid was prepared in a reactor with a volume, and crushed legs were loaded into it in a ratio of 1: 1.2 by weight. The mixture was left for 10 h at a temperature of 25 ° C for hydrolysis, stirring occasionally to ensure maximum contact of the raw material with the acetic acid solution.

To remove hydrolysis products, blood, most of the fat and other impurities, the raw materials were washed twice with tap water on a mesh with a mesh size of 3 mm. Separation of raw materials from wash water was carried out on a batch-type settling centrifuge.

The raw material, finally washed and separated from water, was weighed. For the subsequent extraction of collagen protein from the prepared raw material, a 1.8% aqueous solution of glacial acetic acid was prepared in a ratio of 1: 1.2 to the weight of the raw material, while the pH of the mixture was 4.5. The extraction of raw materials is carried out in tilting boilers with mechanical stirrers, maintaining the temperature of the mixture at 50 ° C for 50 minutes. The resulting extract was fed into a collection tank equipped with a heating jacket, and meat and bone meal was unloaded into containers and used for fodder purposes as a mineral supplement. Next, the extract was purified by filtration on a Druk filter. Preliminarily, a precoated layer of filtered perlite of group B with a thickness of 30 mm was applied to the filter cloth. Filtration was carried out by creating pressure with compressed air in the collector, from which the extract flows to the Druk filter. The purified extract was collected in a collection tank equipped with a heating jacket. During filtration, the temperature of the extract was maintained at 50 ° C. The yield of the extract in the gel state was 80% of the mass of the prepared raw material (cleaned, washed and crushed legs of broiler chickens). At a temperature of the collagen extract from 50 ° C to 37 ° C, it is a colorless, sticky, slightly opalescent liquid, and at room temperature (18-25 ° C), the extract is an elastic, transparent gel that persists for a long time without syneresis. The time taken to obtain the extract was 15 hours.

Example 3 (comparative). Carried out analogously to example 1. But at the stage of hydrolysis, the acetic acid solution is taken in 1.5% concentration and kept for 6 hours at room temperature. This leads to insufficient purification of raw materials and insufficient preparation of raw materials (softening) for subsequent extraction, and with an acetic acid concentration of more than 2.5% and a process duration of more than 10 hours, premature destruction of bone tissue and leaching of calcium, potassium, etc. salts occur, which leads to to reduce the yield of collagen protein from the feedstock (yield less than 50 wt.%).

Example 4 (comparative). Carried out analogously to example 1. But at the stage of collagen protein extraction, the acetic acid solution is taken at 1.4% concentration and the mixture is kept at a temperature of 40 ° C for 45 minutes. In this case, there is an insufficient yield of collagen protein from the feedstock, and at an acetic acid concentration of more than 1.8%, a temperature above 55 ° C and a duration of more than 50 minutes, the collagen protein is denatured and converted into gelatin.

Also, studies were carried out on the possibility of using collagen extract for the local treatment of burn wounds in experimental animals. As a result of the work carried out, it was concluded that “treatment with collagen extracts has a beneficial effect on the course and healing of burn wounds. This is manifested in a decrease in exudative phenomena (hyperemia, edema, migration of leukocytes), in a decrease in the severity or even relief of purulent inflammation. According to the data of histological studies, the specified collagen extracts have sorption and hydration properties. "

In addition, the collagen extract obtained by us from the legs of broiler chickens before their immunological maturation was tested in the beauty institute “Institute of Beauty”. As a result of the study, it was noted that the drug is harmless, the absence of irritating and allergenic effects was stated. It has a positive effect on lipid metabolism, exhibits a water-retaining effect, stimulates an increase in hair follicles and the size of the sebaceous glands, is biologically active. Collagen extract in 5-20% concentration is recommended for use in cosmetic products.

Claims (4)

1. A method of obtaining collagen extract from the legs of broiler chickens, providing for the stages of raw material purification, washing, grinding, hydrolysis and extraction with acetic acid solution, and purification of the extract, characterized in that the hydrolysis is carried out with a 2.0-2.5% solution acetic acid with exposure for 6-10 hours at a temperature of 18-25 ° C, and the extraction of raw materials is carried out with a 1.5-1.8% solution of acetic acid at a temperature of 50-55 ° C for 45-50 minutes at a constant stirring. 2. The method according to claim 1, characterized in that the extract is purified by filtration on a Druk filter with a layer of group B filtered perlite applied to it. 3. A method according to claim 2, characterized in that the pearlite layer is applied with a thickness of 25-30 mm. 4. A method according to claim 2, characterized in that during filtration, the temperature of the extract is maintained within the range of 50-55 ° C.