RU2250047C1 - Food restorative prophylactic product out of cartilaginous tissue of hydrobionts and method for its obtaining - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: the present innovation deals with restorative, nutritive additives for preventing diseases of osseous-muscular system and connective tissue. The suggested restorative product is being an enzymatic-acidic hydrolyzate out of cartilaginous tissue of sharks, or skates, or sturgeon, or salmon fishes, or Cephalopodic mollusks. Moreover, the product contains free hexosamines, chondroitin sulfates, soluble collagen, micro- and macroelements, free amino acids and low-molecular inhibiting agents of metaloproteinases and protein at certain quantitative ratio. The method deals with homogenization of cartilaginous tissue of hydrobionts, extraction with 0.9-2.0% sodium chloride, followed by enzymatic hydrolysis, enzymatic inactivation and separation of the end product. Moreover, enzymatic hydrolysis should be carried out with proteolytic enzymes of microbial or animal origin for 7-8 h. After enzymatic inactivation one should perform lipid precipitation with 0.2-0.3%-chitosan solution in2-4%-acetic acid for 9-15 h. Then residue should be removed, fat-free enzymatic hydrolyzate should be dialyzed against distilled water, the residue undergoes acid hydrolysis with 2-4 M hydrochloric acid solution. Out of acid hydrolyzate it is necessary to isolate hexosamines due to treatment with calcium chloride. Then the residue containing hexosamines, and dialyzed enzymatic hydrolyzate should be mixed and dried at obtaining food restorative prophylactic product. The innovation enables to obtain the product at widened spectrum of biological activity, being, additionally, of antiphlogistic activity.

EFFECT: more improved palatability of the product.

6 ex

Description

The invention relates to the food industry and can be used as a general strengthening, nutritional supplement intended for enhancing the immune system and preventing diseases of the musculoskeletal system and connective tissue, as well as for introducing into food products used for diseases of the musculoskeletal system.

Known products obtained from cartilage and bone tissue of animal origin (patent RU 2082416), cartilage of birds (patent US 5529786), cartilage of fish (patent US 6149946, patent RU 2181292, patent RU 2161002).

A known product (Patent US 6025327, MKI 7 A 61 K 38/39), representing a powdered hydrolyzed collagen type II, is recommended for the correction of the human diet in case of disturbances in the structure of cartilage. The amount of product used is 2000-3000 mg / day daily. The drug does not contain amino sugars with chondroprotective properties.

A known aqueous extract from dried shark cartilage (Application WO 9836760, MKI 6 A 61 K 35/32) containing inhibitors and activators of angiogenesis. An angiogenesis inhibitor is useful for preventing and preventing the growth of tumors. An angiogenesis activator is especially useful for stimulating wound healing.

A known product (Application WO 98/44929, MKI A 61 K 31/70, 38/01) containing proteins of enzymatically hydrolyzed collagen with a molecular weight of 1-300 kDa, which are combined with salts of glucosamine or chondroitin sulfate. Salts of glucosamine and chondroitin sulfate are synthetic analogues and are added additionally. The product is recommended for the prevention of arthrosis.

Known products recommended for arthritis, and methods for their preparation using type II collagen, chondroitin sulfate and glucosamine (US Patent 6162787, MKI 7 A 61 K 38/00). Products contain insoluble native collagen type II in granular form in combination with chondroitin, glucosamine, ascorbate, boron and magnesium. Methods for producing native collagen, type II in granular form are proposed.

Known nutritional supplement (Application WO 9945798, MKI 6 A 23 L 1/304, A 23 J 1/10, A 61 K 35/32), based on bioorganic calcium. The composition is obtained from animal bones, previously removing lipids from the bones by steam treatment or boiling. The bones are then pulverized to form a suspension into which a neutral lipase is introduced for secondary lipid removal and a protease is added to obtain a primary enzymatically decomposed liquid. Papain is introduced into this liquid to obtain a secondary enzymatically decomposed liquid, which is acidified (activated) to form the final product, which is readily absorbed in the human intestine.

A known product, which is an extract of shark cartilage and a method for its production (RF Patent 2181292, MKI A 61 K 35/36), is recommended as an anti-collagenolytic, anti-inflammatory, angiogenic and anti-tumor. A method of obtaining an aqueous extract of shark cartilage includes the following stages: homogenization of cartilage, water extraction, centrifugation of the mixture to separate the extract and its subsequent fractionation, to obtain molecules with a molecular weight of less than and equal to 500 kDa.

A known method and product using a chord of sturgeon fish (Patent US 6149946, MKI A 61 K 35/60). The product is an extract from a sturgeon chord dried in liquid nitrogen, which is suspended in 0.01 M acetic acid. The product is intended to relieve the symptoms of arthritis. For arthritis, in particular rheumatoid arthritis in mammals, including humans, it is recommended to include chord, its extracts and their mixtures in an amount of 1.0 μg - 1.05 g / day.

A known method for producing a complex product (Patent RU 2082416, MKI 6 A 61 K 35/32) containing mucopolysaccharides and collagen from animal raw materials. The method includes grinding raw materials, hydrolysis with a 2.0-8.8% aqueous sodium hydroxide solution for 8-70 hours, depending on the starting tissue, homogenization, acid extraction, isolation and drying of the target product. The extraction is carried out with perchloric acid at a pH of 1.5-2.5, and the target product is isolated from the extract by dialysis against distilled water through a semipermeable membrane. The disadvantage of this method is the duration of the process.

The known product STIMBON-3 and the method for its preparation described in the application for the invention of the Russian Federation 97116345/14, 6 A 61 K 35/32. The product contains bone collagen, products of hydrolysis of glycosaminoglycans of bone tissue, bone non-collagen proteins with a molecular weight of 5-10 kDa with hypoglycemic properties, and novocaine.

The method of obtaining the product consists in the fact that collagen is obtained from the decalcified dissociatively extracted bone tissue matrix and the products of acid hydrolysis of glycosaminoglycans are extracted from the dissociative bone extract with a relative molecular weight of 5-10 kDa protein of neutral and basic nature, mixed in the specified proportion, collagen is added . The product Stimbon-3 is recommended for degenerative-dystrophic lesions of the articular cartilage. The disadvantage of this product is the additional introduction of collagen.

A means of preventing degenerative-dystrophic changes in cartilage tissue is known (Patent RU 2201757, MKI A 61 K 35/32, 38/08, A 61 F 9/00, A 61 P 19/02), which is a composition comprising calcium, phosphorus and a mixture of amino acids, which in its composition has uronic acids and hexosamines, while the composition per 100 g of product is:

uronic acids 2.8-3.2 g

hexosamines 4.8-5.2 g

calcium 42.0-45.0 g

phosphorus 27.0-30.0 g

magnesium 2.6-2.8 g

amine nitrogen as free amino acids and

oligopeptides rest

The production method includes the following steps: bone matrix decalcification, acid hydrolysis, mixing the components and drying them. The hydrolyzate of glycosaminoglycans of articular cartilage is obtained from the cartilage of the purified cattle trachea, for which they are subjected to acid hydrolysis in hydrochloric acid until completely dissolved, then decalcified with hydrochloric acid and precipitated with calcium oxide from the purified and ground cattle bones, a bone mineral is obtained, after which mixing the mineral phase with cartilage hydrolyzate and drying.

In General, the disadvantages of the known methods include harsh conditions of hydrolysis, the additional introduction of synthetic components.

The closest technical solution to the technical nature of both the product and the method are the product and the method for its preparation described in RF patent No. 2161002, 7 A 23 L 1/325, A 61 K 35/60.

This product has immunostimulating and antioxidant activities and is an enzymatic hydrolyzate of the cartilaginous tissue of sharks of the species Lamna ditropis, Halaerurus buergeri, Galeocerdo cuvier, Squalus acanthias, Somniosus antarcticus, Somniosus paciflcus and contains no less than 20% by weight free amino acids in dry matter. including aromatic amino acids - tryptophan, tyrosine and phenylalanine, the amount of which is at least 22% of the total number of amino acids, diamino acids - lysine and arginine - at least 19%, dicarboxylic amino acids - asparagine and glutamine - at least 17%, and glycine not less than 6% and taurine not less than 1.5%. The composition of the product includes hyaluronic acid in an amount of 2.1-5.5 mg / g, not more than 15% protein and 0.4-1.5 mg / g of free amino sugars by weight of the dry preparation.

The disadvantage of the prototype - this product from the cartilaginous tissue of sharks is not a wide range of biological activity, high content of sodium chloride, low content of hexosamines.

The process of obtaining this product (prototype) from the cartilage tissue of sharks of the species Lamna ditropis, Halaerurus buergeri, Galeocerdo cuvier, Squalus acanthias, Somniosus antarcticus, Somniosus pacificus includes the following stages: raw materials - fresh or dry sharks cartilage - crush and extract 2 0% sodium chloride solution for 10-18 hours at 5-10 ° C, enzymatic hydrolysis is carried out using proteolytic enzyme preparations at 30-45 ° C for 15-24 hours, inactivation of enzymes is carried out by heating to 70-80 ° C in for 15-20 minutes, the subsequent selection of the target product is carried out by filtration, drying is carried out by sublimation or spray method. The yield of the final product is 6-8%.

Enzymatic hydrolysis is carried out using a proteolytic enzyme preparation isolated from pyloric appendages of Far Eastern salmon (pylorin) with an activity of 100-200 U / g in an amount of 1-4% of the mass of ground cartilage, or a proteolytic enzyme preparation isolated from king crab hepatopancreas with an activity of 150 -250 U / g in an amount of 0.5-3% of the mass of ground cartilage, or using megaterina with an activity of 200-400 U / g in an amount of 1-4% of the mass of ground cartilage.

The disadvantages of the prototype - a method of obtaining a product from the cartilaginous tissue of sharks is the long duration of enzymatic hydrolysis, a high content of sodium chloride and a low yield of the target product.

The problem solved by the invention is the expansion of the range of food fortifying prophylactic products from cartilaginous tissue of aquatic organisms, obtaining a product with a wider spectrum of biological activity due to the preparation of a complex preparation that additionally has anti-inflammatory activity, improving the taste of a product containing soluble active ingredients that are easily absorbable in a soluble form in the human intestines.

The essence of the invention lies in the fact that the food fortifying prophylactic product from cartilaginous tissue of aquatic organisms, having immunostimulating, antioxidant, anti-inflammatory activities, is an enzymatic-acid hydrolyzate from cartilaginous tissue of aquatic organisms and contains:

free hexosamines - 0.8-6.5%,

chondroitin sulfates - 5.8-7.0%,

soluble collagen - 3.8-13.8%,

micro and macro elements, of which

40-60% - calcium, 5.8-17% - magnesium - 10-11%,

free amino acids - 10-18%,

low molecular weight inhibitors

metalloproteinases with mm 0.3-10 kDa - 0.1-1.5%,

protein - 42-55%.

It is known that hexosamines and chondroitin sulfates are precursors of articular cartilage macromolecules, and their introduction into the diet causes a stimulating effect and accelerates the regeneration of cartilage tissue through the use of ready-made building material and the ability to accumulate in the foci of inflammation. Their activity depends on the presence in their structure of at least one negatively charged carboxyl or sulfate group, which provides hydrophilicity and surface-active properties. Low molecular weight metalloproteinase inhibitors prevent an increase in the activity of enzymes involved in the inflammatory process. Free amino acids are necessary and beneficial for the repair of connective tissues. Calcium contributes to the normal growth and development of the body, serves as a building material for bones, is necessary for the functioning of the nervous, muscle and bone systems. Magnesium, interacting with calcium, affects the permeability of membranes, contributing to an increase in the level of calcium in the body.

New in the object of the invention, the product is that it is obtained complex, possessing immunostimulating, antioxidant, anti-inflammatory activity, purified from lipids, in a soluble form and with a low content of sodium chloride, a nutritionally strengthening prophylactic product from cartilaginous tissue of hydrobionts.

The content of hexosamines and chondroitin sulfates was determined spectrophotometrically, according to the methodology of pharmacopoeial article No. 42-1286-99. The amount of collagen was determined by the method of Krylova N.N. and Lyaskovsky Yu.N. "Physicochemical methods for the study of animal products." M .: Food industry, 1965, p. 49-55. The study of the elemental composition of cartilage tissue preparations was carried out by atomic absorption method on a Nippon Jarrel Ash flame emission spectrophotometer model AA-855. The protein content was determined by the amount of nitrogenous substances (in terms of protein) using the Kjeldahl micromethod using the Kjeltec auto 10 SO Analyzer instrument. The inhibitory activity of the components of the cartilage tissue was determined by Pivnenko T.N., Klychkova G.Yu. // Bulletin of TINRO, 2001, V.129, p. 74-81.

Antioxidant activity was determined by the accumulation of malondialdehyde in blood serum in the presence of an exogenous oxidation system. The amount of malondialdehyde accumulated during the day of incubation in the presence of 0.25 M [Fe ²⁺ / Fe ³⁺ ] (accelerated oxidation catalyst) in normal anti-Rhesus human blood serum was determined at a concentration of 2 mg / l of serum. Serum with the addition of an oxidation catalyst was used as a control. The content of malondialdehyde in the presence of products from cartilage of hydrobionts amounted to 0.014-0.0175 mg / ml of blood serum. In the control sample into which the oxidation catalyst was introduced, but cartilage preparations were absent, the content of malondialdehyde was 0.070-0.072 mg / ml.

The immunostimulating effect of the product was determined and evaluated in the following tests:

determination of the effect of the product on the phagocytic activity of polymorphonuclear leukocytes (PMNL) and macrophages in an in vivo and in vitro experiment;

determination of the effect of the product on the number of antibody-forming cells in the spleens of mice immunized with sheep red blood cells;

determining the effect of the product on the restoration of rosette-forming cells from guinea pig spleens The essence of the method is to isolate cells, treat them with trypsin to violate the surface structure. The ability of cells to recover in the presence and absence of the test drug from the cartilaginous tissue of hydrobionts was further evaluated. The recovery rate of rosette-forming cells, expressed as a percentage, was taken as the main immunological indicator; it is at least 15% for the preparations obtained compared with the control.

The anti-inflammatory activity of products from cartilaginous tissue of aquatic organisms was determined on a model of infectious-allergic pseudotuberculous arthritis in rabbits. Experimental arthritis was caused by antigenic irritation of tissues of the right knee joint of animals previously (for 7 days) sensitized by culture of pseudotuberculosis microbe strain 512. Animals were sensitized by subcutaneous injection of a suspension of bacteria in the right thigh at a dose of 2 × 10 ⁷ microbial cells. After 7 and 14 days, rabbits were injected with corpuscular pseudotuberculosis antigen at a dose of 10 ⁷ -10 ⁸ mk into the cavity of the knee joint under aseptic conditions. Infected animals were divided into two groups: the experimental group (20 rabbits) received a once-daily test preparation dissolved in distilled water daily for 14 days in an amount of 22.5 mg, the control group (6 rabbits) did not receive treatment.

During an experimental-morphological study of the anti-inflammatory effect of a product from the cartilaginous tissue of aquatic organisms, it was noted that the animals of the experimental group were active, gained weight, had a smooth coat without loss. In the control group, abundant hair loss and inconsistent claudication, roundness of the configuration of the knee joint, small-pointed hemorrhages in the joint bag, and the presence of synovial fluid in the joint cavity were observed.

Microscopic examination revealed changes in the synovial membrane of the knee joint. In animals of the experimental group, after consumption of the product, a slight disengagement of the connective tissue, slight hyperemia of the vessels, single hemorrhages, residual reactive-inflammatory changes were observed. In the control group, synovia edema, swelling of collagen fibers, moderate expansion and plethora of vessels, fibrinoid swelling of the walls of the vessels, diapedetic hemorrhages were noted. Perivascular and weak diffuse infiltration of the synovial membrane by macrophage lymphoid cells with an admixture of polymorphonuclear leukocytes, eosinophils and plasma cells indicates the development of the inflammatory process.

At the level of macroscopic changes in the internal organs of the control animals, cyanotic coloration of the lung parenchyma, necrotic foci along the edge and in the liver parenchyma were revealed. In animals of the experimental group, only single foci were observed in the repair stage.

Microscopic changes in internal organs (liver, spleen, lungs, lymph nodes) indicate mild necrotic changes, the presence of small foci of bronchopneumonia and dystrophic changes in hepatocytes in the control. In animals of the experimental group, the above changes occurred against the background of hyperplasia of bronchial associated lymphoid tissue, moderate lymphoid-macrophage infiltration in the portal tracts of the liver, hyperplasia of the B and T-dependent zones of the spleen and lymph nodes.

Thus, a study of anti-inflammatory activity suggests that the product from the cartilaginous tissue of aquatic organisms has a positive effect and reduces the morphological manifestations of experimental infectious-allergic pseudotuberculous arthritis in rabbits. The product also has a stimulating effect, causing an improvement in the general condition of animals and the development of tissue immunomorphological reactions.

The essence of the invention-method lies in the fact that the claimed food fortifying prophylactic product from cartilage is obtained as follows: raw materials - fresh or dry cartilage tissue of sharks, or stingrays, or sturgeon and salmon fish, or cephalopods (squid, octopus) are crushed and extracted with 0.9-2.0% sodium chloride solution for 10-18 hours at 5-10 ° C, enzymatic hydrolysis is carried out using a proteolytic enzyme of microbial or animal origin at 30-40 ° C for 7-8 hours, inactivation the enzyme is carried out by heating the mixture to 70-80 ° C for 15-20 minutes, the subsequent lipid precipitation is carried out with a 0.2-0.3% solution of chitosan in 2-4% acetic acid for 9-15 hours at 5-10 ° C , then the mixture is centrifuged, or passed through a metal filter. Next, the purified enzymatic hydrolyzate is dialyzed against distilled water to obtain a solution containing soluble collagen, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, free chondroitin sulfates, a mixture of amino acids. The precipitate is subjected to acid hydrolysis in 2-4 M hydrochloric acid, followed by precipitation of hexosamines with calcium chloride. The precipitate containing hexosamines and the dialyzed enzymatic hydrolyzate are mixed and dried to obtain the desired product.

A new method for producing a food-strengthening prophylactic product from cartilage of aquatic organisms is the introduction of stages of purification of the enzymatic hydrolyzate by precipitation of lipids with 0.2-0.3% solution of chitosan in 2-4% acetic acid for 9-15 hours at 5-10 ° C. and subsequent dialysis of the enzymatic hydrolyzate against distilled water, contributing to the improvement of the taste of the product. Used in the proposed method, proteolytic enzymes of microbial and animal origin, such as protomex, pancreatin, pylorine and proteolytic enzyme preparation from hepatopancreas king crab, allow you to save vital amino acids in the final product, such as tryptophan, cysteine, as well as low molecular weight inhibitors of metalloprotein, acid hydrolysis of raw materials are destroyed, and the additionally introduced step of acid hydrolysis of the precipitate allows increasing the content of hexosamines in odukte, and as a result - the emergence of anti-inflammatory activity and an increase in the final yield.

In the proposed method, in contrast to the prototype, a complex preparation is obtained containing the products of enzymatic acid hydrolysis of the cartilaginous tissue of sturgeon, or salmon fish, or sharks, or stingrays, or cephalopods (squids, octopuses), namely the soluble form of hexosamines, chondroitin sulfates, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, collagen, Ca and Mg, easily absorbed in the human intestine.

The invention is illustrated by the following examples.

Example 1

The food fortifying prophylactic product from shark cartilage is an enzymatic acid hydrolyzate and contains:

free hexosamines in an amount of 3.45%,

chondroitin sulfates - 6.8%,

soluble collagen - 4.0%,

micro and macro elements, of which

40% is calcium, 6.3% is magnesium - 11.0%,

free amino acids - 15.0%,

low molecular weight metal inhibitors

proteinases with a molecular weight of 0.3-10 kDa - 0.3%,

protein - 51.45%,

moisture - 8%.

This product is prepared as follows: 1 kg of shredded cartilage of sharks was extracted into 4 l of a 1% sodium chloride solution for 15 hours at 10 ° C, then the extract was heated to 38 ° C, the pH of the solution was adjusted to 8.0, and the enzyme preparation was added "Pancreatin" in the amount calculated in units of activity per mass of feedstock, previously dissolved in 0.2 l of the same solution, was thoroughly mixed and kept for 7 hours at 38 ° C, stirring was carried out periodically. The inactivation of the enzyme was carried out by heating to 80 ° C for 20 min. Lipid deposition was carried out with a 0.3% solution of chitosan in 2% acetic acid for 10 hours at 8 ° C. The solution was centrifuged at 3000 rpm for 30 minutes. Next, the purified enzymatic hydrolyzate was dialyzed against distilled water to obtain a solution containing soluble collagen, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, free chondroitin sulfates, and a mixture of amino acids. The precipitate was subjected to acid hydrolysis in 4 M hydrochloric acid, followed by precipitation of hexosamines with calcium chloride. The precipitate containing hexosamines and the dialyzed enzymatic hydrolyzate were mixed and freeze-dried. The yield of the drug was 10.1%.

Antioxidant activity was 20.7%, which corresponds to a MDA content of 0.0145 mg / ml serum.

The ROC recovery index was 15.6%.

Example 2

The food fortifying prophylactic product from the cartilage of sturgeon tissue is an enzymatic acid hydrolyzate and contains:

free hexosamines in an amount of 3.36%,

chondroitin sulfates - 6.0%,

soluble collagen - 13.5%,

micro and macro elements, of which

58% is calcium, 9.8% is magnesium - 10.0%,

free amino acids - 14.0%,

low molecular weight metal inhibitors

proteinases with a molecular weight of 0.3-10 kDa - 1.3%,

protein - 43.84%,

moisture - 8%.

This product is prepared as follows: 1 kg of ground sturgeon cartilage was extracted into 4 L of a 2% sodium chloride solution for 12 hours at 6 ° C, then the extract was heated to 37 ° C, the pH of the solution was adjusted to 8.0, and an enzyme preparation was added. Pilorin, from pyloric appendages of salmon fish, in an amount calculated in units of activity per mass of feedstock, previously dissolved in 0.2 l of the same solution, was thoroughly mixed and kept for 7 hours at 37 ° C, stirring was carried out periodically. The inactivation of the enzyme was carried out by heating to 70 ° C for 20 min. Lipid precipitation was carried out with a 0.3% solution of chitosan in 2% acetic acid for 15 hours at 5 ° C. The solution was centrifuged at 3000 rpm for 30 minutes. Next, the purified enzymatic hydrolyzate was dialyzed against distilled water to obtain a solution containing soluble collagen, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, free chondroitin sulfates, and a mixture of amino acids. The precipitate was subjected to acid hydrolysis in 3 M hydrochloric acid, followed by precipitation of hexosamines with calcium chloride. The precipitate containing hexosamines and the enzymatic hydrolyzate were mixed and dried in a drum type dryer. The yield of the drug was 20%.

Antioxidant activity was 25%, which corresponds to a MDA content of 0.0175 mg / ml serum.

The ROC recovery index was 17.9%.

Example 3

The food fortifying prophylactic product from the cartilaginous tissue of salmon fish is an enzymatic acid hydrolyzate and contains:

free hexosamines in an amount of 1.1%,

chondroitin sulfates - 6.0%,

soluble collagen - 5.9%,

micro and macro elements, of which

42% is calcium, 6.2% is magnesium - 10.0%,

free amino acids - 16.0%,

low molecular weight metal inhibitors

proteinases with a molecular weight of 0.3-10 kDa - 0.98%,

protein - 53.02%,

moisture - 7%.

This product is prepared as follows: 1 kg of crushed cartilaginous tissue of salmon fish was extracted into 4 l of a 0.9% sodium chloride solution for 18 hours at 5 ° C, then the extract was heated to 35 ° C, the pH of the solution was adjusted to 8.0 and the enzyme preparation isolated from hepatopancreas of king crab was added in an amount calculated in units of activity per mass of feedstock previously dissolved in 0.2 l of the same solution, thoroughly mixed and kept for 7.5 h at 35 ° C, stirring was carried out periodically. The inactivation of the enzyme was carried out by heating to 80 ° C for 20 min. Lipid precipitation was carried out with a 0.2% solution of chitosan in 3% acetic acid for 13 hours at 6 ° C. The solution was passed through a filter cloth. Next, the purified enzymatic hydrolyzate was analyzed against distilled water to obtain a solution containing soluble collagen, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, free chondroitin sulfates, a mixture of amino acids. The precipitate was subjected to acid hydrolysis in 2 M hydrochloric acid, followed by precipitation of hexosamines with calcium chloride. The precipitate containing hexosamines and the dialyzed enzymatic hydrolyzate were mixed and freeze-dried. The yield of the drug was 12%.

Antioxidant activity was 20%, which corresponds to a MDA content of 0.014 mg / ml serum.

The ROC recovery index was 19.5%.

Example 4

The food fortifying prophylactic product from squid cartilage is an enzymatic acid hydrolyzate and contains:

free hexosamines in an amount of 0.99%,

chondroitin sulfates - 6.0%,

soluble collagen - 8.4%,

micro and macro elements, of which

55% is calcium, 16% is magnesium - 10.0%,

free amino acids - 18.0%,

low molecular weight metal inhibitors

proteinases with a molecular weight of 0.3-10 kDa - 1.2%,

protein - 47.41%,

moisture - 8%.

This product is prepared as follows: 1 kg of crushed squid cartilage was extracted into 4 l of a 1.2% sodium chloride solution for 10 hours at 7 ° C, then the extract was heated to 30 ° C, the pH of the solution was adjusted to 8.0, and the Protomex enzyme preparation was added in an amount calculated in terms of activity per weight of the feedstock, previously dissolved in 0.2 L of the same solution, thoroughly mixed and kept for 8 hours at 30 ° C, stirring was carried out periodically. The inactivation of the enzyme was carried out by heating to 80 ° C for 20 min. Lipid precipitation was carried out with a 0.2% solution of chitosan in 4% acetic acid for 12 hours at 9 ° C. The solution was passed through a filter with a metal filter. Next, the purified enzymatic hydrolyzate was dialyzed against distilled water to obtain a solution containing soluble collagen, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, free chondroitin sulfates, and a mixture of amino acids. The precipitate was subjected to acid hydrolysis in 2 M hydrochloric acid, followed by precipitation of hexosamines with calcium chloride. The precipitate containing hexosamines and the dialyzed enzymatic hydrolyzate were mixed and dried on a drum type dryer. The yield of the drug was 9.75%.

Antioxidant activity was 22.1%, which corresponds to a MDA content of 0.0155 mg / ml serum.

The ROC recovery index was 22.4%.

Example 5

The food fortifying prophylactic product from the cartilage of the octopus is an enzymatic acid hydrolyzate and contains:

free hexosamines in an amount of 1.0%,

chondroitin sulfates - 6.1%,

soluble collagen - 9.1%,

micro and macro elements, of which

51% is calcium, 15% is magnesium - 11.0%,

free amino acids - 17.0%,

low molecular weight metal inhibitors

proteinases with a molecular weight of 0.3-10 kDa - 1.0%,

protein - 47.8%

moisture - 7%.

This product is prepared as follows: 1 kg of ground octopus cartilage was extracted into 4 l of a 1.1% sodium chloride solution for 11 hours at 9 ° C, then the extract was heated to 33 ° C, the pH of the solution was adjusted to 8.0, and the enzyme preparation isolated from hepatopancreas of king crab was added in an amount calculated in units of activity per mass of feedstock previously dissolved in 0.2 l of the same solution, thoroughly mixed and kept for 7.5 h at 33 ° C, stirring was carried out periodically. The inactivation of the enzyme was carried out by heating to 80 ° C for 20 min. Lipid precipitation was carried out with a 0.2% solution of chitosan in 3% acetic acid for 11 hours at 7 ° C. The solution was centrifuged at 3000 rpm for 30 minutes. Next, the purified enzymatic hydrolyzate was dialyzed against distilled water to obtain a solution containing soluble collagen, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, free chondroitin sulfates, and a mixture of amino acids. The precipitate was subjected to acid hydrolysis in 2 M hydrochloric acid, followed by precipitation of hexosamines with calcium chloride. The precipitate containing hexosamines and the dialyzed enzymatic hydrolyzate were mixed and freeze-dried. The drug yield was 10.5%.

Antioxidant activity was 22.1%, which corresponds to a MDA content of 0.0155 mg / ml serum.

The ROC recovery index was 19.8%.

Example 6

The food fortifying prophylactic product from stingray cartilage tissue is an enzymatic acid hydrolyzate and contains:

free hexosamines in an amount of 6.1%,

chondroitin sulfates - 6.5%,

soluble collagen - 4.9%,

micro and macro elements, of which

43% is calcium, 6.4% is magnesium - 11.0%,

free amino acids - 13%,

low molecular weight metal inhibitors

proteinases with a molecular weight of 0.3-10 kDa - 0.4%,

protein - 50.1%

moisture - 8%.

This product is prepared as follows: 1 kg of crushed stingray cartilage was extracted into 4 l of a 1.5% sodium chloride solution for 16 hours at 8 ° C, then the extract was heated to 39 ° C, the pH of the solution was adjusted to 8.0 and added enzyme preparation "Pilorin" isolated from pyloric appendages of salmon fish, in an amount calculated in units of activity per mass of feedstock, previously dissolved in 0.2 l of the same solution, was thoroughly mixed and kept for 8 hours at 39 ° C, stirring carried out periodically. The inactivation of the enzyme was carried out by heating to 70 ° C for 20 min. Lipid precipitation was carried out with a 0.2% solution of chitosan in 4% acetic acid for 9 hours at 10 ° C. The solution was centrifuged at 3000 rpm for 30 minutes. Next, the purified enzymatic hydrolyzate was dialyzed against distilled water to obtain a solution containing soluble collagen, low molecular weight metalloproteinase inhibitors with a molecular weight of 0.3-10 kDa, free chondroitin sulfates, and a mixture of amino acids. The precipitate was subjected to acid hydrolysis in 4 M hydrochloric acid, followed by precipitation of hexosamines with calcium chloride. The precipitate containing hexosamines and the dialyzed enzymatic hydrolyzate were mixed and freeze-dried. The yield of the drug was 14%.

Antioxidant activity was 21.4%, which corresponds to a MDA content of 0.015 mg / ml serum.

The ROC recovery index was 15.9%.

Claims (3)

1. Food fortifying prophylactic product from cartilaginous tissue of aquatic organisms, characterized in that it is an enzymatic acid hydrolyzate of cartilaginous tissue of sharks, or stingrays, or sturgeon, or salmon fish, or cephalopods (squid, octopus) and contains: free hexosamines in an amount of 0.8-6.5%, chondroitin sulfates - 5.8-7.0%, soluble collagen - 3.8-13.8%, micro and macro elements, of which 40-60% is calcium, 5.8-17% - magnesium - 10-11.0%, free amino acids - 10-18%, low molecular weight metal inhibitors proteinases with a molecular weight of 0.3-10 kDa - 0.1-1.5%, protein - 42-55%. 2. A method of producing a food-strengthening prophylactic product from hydrobiont cartilage tissue, including tissue homogenization, extraction with 0.9-2.0% sodium chloride, enzymatic hydrolysis, enzyme inactivation and separation of the final product, characterized in that the shark cartilage tissue is used as a raw material or stingrays, or sturgeon and salmon fish, or cephalopods (squid, octopus), enzymatic hydrolysis is carried out by proteolytic enzymes of microbial or animal origin for 7-8 h, and after inactivation of the enzymes, lipids are precipitated with a 0.2-0.3% solution of chitosan in 2-4% acetic acid for 9-15 hours, then the precipitate is separated, the fat-free enzymatic hydrolyzate is dialyzed against distilled water, the precipitate is subjected to acid hydrolysis 2 With a 4 M hydrochloric acid solution, hexosamines are isolated from the acid hydrolyzate by treatment with calcium chloride, after which the precipitate containing hexosamines and the dialyzed enzymatic hydrolyzate are mixed and dried to obtain a nutritional restorative preventative about the product. 3. The method according to claim 2, characterized in that the enzymatic hydrolysis of the cartilage tissue is carried out using a proteolytic enzyme preparation from the Protomex series, or Pancreatin, or Pilorin isolated from pyloric appendages of salmon fish, or a proteolytic enzyme preparation, Kamchatka crab isolated from hepatopancreas in amounts corresponding to units of activity per mass of feedstock.