RU2317714C2 - Biologically active additive based on cyst, or nauplii, or adult persons of naxillopod artemia salina for topic or oral administering - Google Patents

Abstract

FIELD: food or cosmetic biologically active additive, medicine, cosmetic, food processing industry.

SUBSTANCE: claimed additive contains lipid and protein-amino acid fraction from extract of cyst, or nauplii, or adult persons of naxillopod Artemia salina in ratio from 1000:1 to 1:1.

EFFECT: biologically active additive with controlled content of proteins and amino acids and reduced allergenic reactions.

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Description

The invention relates to food or cosmetic dietary supplements and can be used in the food industry or cosmetics.

Despite the achievements of modern science, the problem of the deficiency of complex additives balanced in vitamins, minerals and amino acids remains relevant. To maintain these indicators, food should contain not only the right amount of protein, but also the required vitamin and amino acid composition. The value of Artemia cysts is determined by its chemical composition, which is characterized by a fairly high content of proteins, fats, essential amino acids and fatty acids, vitamins, hormones, and other biologically active compounds. Artemia is common throughout the globe. In Russia, it lives in salt water bodies of the coast of the southern seas, lakes of the Caucasus, Kyrgyzstan, Western and Eastern Siberia. The water area of the studied reservoirs of Western Siberia alone is 2.1 thousand km ² , of which 1.1 thousand km ² in the Altai Territory.

It is known to use whole eggs of the gill leg Artemia salmon (L) in the form of a dry powder as a feed additive for birds, as well as a feed additive, which use whole eggs of the gill leg gem of Artemia (Artemia salina (L) in the form of a dry powder (up to 10-15 wt.%) With the addition of grain or other feed (up to 100 wt.%) Used for feeding laying hens (RF Patent No. 2158519, IPC A23K 1/16, 1/00; publ. 10.11.2000, )

However, whole brine shrimp eggs are covered with a chitinous membrane and can be absorbed only by the body of birds, which grind this product in the corresponding section of the alimentary canal. Using whole Artemia eggs as a dietary supplement in a person’s diet or as a component of a medicinal, cosmetic or balneotherapy drug is inefficient due to the poor digestibility of whole Artemia cysts in this form.

Known food dietary supplement, which is a purified concentrate of omega-3 polyunsaturated fatty acids, obtained from the extract of crustaceans Artemia salina (US Patent No. 5340594, IPC A23D 9/00, publ. 23.08.1994).

The disadvantage of this biologically active additive is the absence of many useful components in it that are contained in the lipid-vitamin complex of Artemia cysts (polyunsaturated fatty acids omega-6, squalene, astaxanthin, and others), which limits its biological activity and scope.

A dry form of a dietary supplement based on Artemia salina cyst is known, which is crushed to an ultrafine state with a particle size of not more than 20 μm and a moisture content of not more than 5.0 wt.%. The dry form of the treatment-and-prophylactic preparation is a tablet made in the form of a monolithic disk-shaped solid body from Artemia cyst powder, or a cylindrical hollow detachable capsule filled with Artemia cyst powder, or a gas-tight bag filled with Artemia cyst powder in vacuum (Certificate of Useful RF model No. 45270, IPC A61K 35/56, publ. 10.05.2005).

Known use in the composition of solid rennet cheese food biological active additives based on decapsulated cysts of brine shrimp Artemia salina in an amount of 0.25-1.0 wt.% (Patent application of the Russian Federation No. 2003112517, IPC A23C 19/068, publ. 27.11.2004 )

Known compositions of creams, ointments, emulsions based on the extract of crustaceans Artemia salina, used in cosmetics for the prevention and treatment of skin and hair (French Patent No. 2817748, IPC A61K 35/56, 7/48, publ. 06/14/2002).

The closest analogue (prototype) is also known, which is a biologically active additive for medical and cosmetic preparations, which has an antiproliferative effect and is an aqueous or alcoholic extract of eggs (cysts) of crustaceans Artemia salina (US Patent No. 6342254, IPC АКК 35/78, 35 / 12, published on January 29, 2002). The tool is proposed to be used in medicine and cosmetics for the treatment and prevention of skin diseases.

The disadvantage of these analogues, including the prototype, is that the liquid extract of eggs (cysts) or adult crustaceans Artemia salina contains in addition to the lipid-vitamin fraction a large amount of proteins, peptides and amino acids, which when added to food products or used in medicine or cosmetics can cause allergic reactions of the body, which affect 20 to 40% of children and up to 15% of the adult population of Russia and other countries. The above factors narrow the scope of these highly allergenic dietary supplements in the food industry, as well as in medicine and cosmetics.

The technical result of the invention is the creation of a biologically active additive based on an extract of cysts, or nauplii, or adult Artemia salina crustaceans with controlled protein and amino acid content by dosing a protein-amino acid fraction into a biologically active additive containing a lipid fraction of this extract.

The specified technical result is achieved by the creation of a biologically active additive (BAA) containing lipid and protein-amino acid fractions from the extract of crushed cysts, or nauplii, or adult Crayfish Artemia salina in a ratio of from 1000: 1 to 1: 1.

The protein-amino acid fraction of the aqueous-alcoholic extract of cyst of Artemia salina is characterized by a rather high content of proteins, essential and non-essential amino acids, and other biologically active compounds.

The lipid fraction of the aqueous-alcoholic extract of cysts of Artemia salina contains polyunsaturated fatty acids, vitamins, steroids and hormone-like substances, growth factors, and other fat-soluble biologically active compounds.

Dosed introduction of a protein-vitamin fraction (from 1000: 1 to 1: 1) into the lipid fraction, depending on the purpose of the final product, ensures a reduction in allergic reactions in consumers of this product.

Example 1. The technology of obtaining lipid and protein-amino acid fractions from the extract of Artemia cysts

Eggs (cysts), nauplii and adult crustaceans Artemia salina are obtained from lakes in Western Siberia, repeatedly washed in fresh water from impurities, salts and low-quality eggs or nauplii. At the stage of washing, whole cysts, nauplii, or adult Artemia crustaceans can be additionally treated with detergents. The wet product is dried in an air stream with a moisture content of not more than 5 wt.%, At a temperature not exceeding 50 ° C. Next, the powder is ground in a vortex mill developed by Vortex Technologies, Novosibirsk (RF patent N 2057588, MKI V02C 19/06, publ. 04/10/96), which is a gas-dynamic mill using compressed gas energy to accelerate crushed particles of the product in a vortex flow to an ultrafine state with a particle size of not more than 20 microns and a moisture content of not more than 5.0 wt.%. As a grinder can also serve as a ball or disk mill or other technical means that provide the appropriate dispersed composition of the product without compromising its useful properties. In accordance with examples 1 and 2, a biologically active additive is obtained from raw materials.

To do this, take 1 kg of Artemia cyst powder is extracted with a water-ethanol mixture (1: 2) in a volume ratio of 1: 6 (raw material: solvent) at room temperature and stirring for 1.0-2.0 hours. Then the mass is filtered through a calico bag on a suction filter and washed in small portions with the same mixture of solvents.

The resulting filtrate is defended until the complete formation of two layers of liquids. The lower lipid layer is poured into a container, followed by packing into bottles of the finished product. If necessary, in order to remove alcohol from the product, the lower lipid layer is poured into the evaporator through a coarse calico sack and the alcohol is completely removed in vacuum. The resulting brown product is a low-melting lipid complex (melting point 50 ° C). Yield 3.7% of the dry weight of raw materials.

The upper water-alcohol layer is also poured into a container, followed by packing into product bottles. To remove alcohol, the upper water-alcohol layer is also poured into the evaporator and the main part of the alcohol is removed in vacuo. If necessary, the resulting solution is dried, the remainder is a cream-colored powder, including a concentrate of amino acids and peptides. In addition, this product contains a small amount of carbohydrates, phospholipids. Yield 7.0% of the dry weight of the feed.

Example 2. The technology of obtaining lipid and protein-amino acid fractions from an extract of nauplii or adult Artemia

Nauplii and adult crustaceans Artemia salina are also mined from lakes in Western Siberia, repeatedly washed in fresh water from impurities, salts and low-quality eggs or nauplii. At the stage of washing, whole cysts, nauplii, or adult Artemia crustaceans can be additionally treated with detergents. The wet product is dried in an air stream with a moisture content of not more than 5 wt.%, At a temperature not exceeding 50 ° C. Next, the powder is ground in a vortex mill, developed by the company "Vortex Technologies", Novosibirsk (RF patent No. 2057588, MKI V02C 19/06, publ. 04/10/96), which is a gas-dynamic grinder that uses the energy of compressed gas to accelerate crushed particles of the product in a vortex flow to an ultrafine state with a particle size of not more than 20 microns and a moisture content of not more than 5.0 wt.%. As a grinder can also serve as a ball or disk mill or other technical means that provide the appropriate dispersed composition of the product without compromising its useful properties. In accordance with examples 1 and 2, a biologically active additive is obtained from raw materials.

To do this, take 1 kg of powder of nauplii or adults Artemia is extracted with a water-ethanol mixture (1: 2) in a volume ratio of 1: 5 (raw materials: solvent) at room temperature and stirring for 1.0-2.0 hours. Then the mass is filtered through a calico bag on a suction filter and washed in small portions with the same mixture of solvents (0.2 volume 3 times with respect to the obtained volume).

The resulting filtrate is defended until the complete formation of two layers of liquids. The lower lipid layer is poured into a container, followed by packing into bottles of the finished product. To remove alcohol from the product, the lower lipid layer is poured into the evaporator through a coarse calico sack and the alcohol is completely removed in vacuum. The resulting brown product is a low-melting lipid complex (melting point 50 ° C). Yield 5.0% of the dry weight of the feed.

The upper water-alcohol layer is also poured into a container, followed by packing into product bottles. To remove alcohol, the upper water-alcohol layer is also poured into the evaporator and the main part of the alcohol is removed in vacuo. The resulting solution is dried, the remainder is a creamy protein complex powder, is a concentrate of amino acids and peptides. In addition, this product contains a small amount of carbohydrates, phospholipids. Yield 8.0% of the dry weight of the feed.

Further, the obtained products (lipid-vitamin and protein-amino acid fractions) are mixed in a ratio of from 1000: 1 to 1: 1 depending on the purpose and can be used as a dietary supplement or component of a pharmaceutical or cosmetic.

A food or pharmaceutical product in dry form is prepared in a powder mixer. The mixer is a container with a spherical bottom having an axis of rotation inclined to the horizontal. The mixing efficiency is ensured by inert bodies of a spherical shape, significantly smaller than the bottom of the tank of radius of curvature, and significantly larger specific gravity than the specific gravity of the mixed components of the material. Inert bodies prevent aggregation of particles and provide mixing with particles of any dispersion in a small gap between the surface of inert bodies and the bottom of the tank. The number of inert bodies ensures their location in only one layer to avoid their active collision and, as a result, to avoid overheating of the mixed materials.

Example 3. Obtaining a pharmaceutical preparation

The following components of the pharmaceutical preparation (wt.%) Are introduced into the mixer:

lipid and protein-amino acid fractions extract of crushed cysts, or nauplii, or adult crustacean Artemia salina in the ratio of 1000: 1 1.0-10.0 a mixture of chalk with starch (2: 1) the rest is up to 100%

The mixture is then passed through a tablet press.

Example 4. Obtaining a nutritional supplement

The following components of the food product (wt.%) Are introduced into the mixer:

lipid and protein-amino acid fractions extract of crushed cysts, or nauplii, or adult crustacean Artemia salina in the ratio 1: 1 0.1-1.0 salt the rest is up to 100%

Next, the resulting product is Packed.

Example 5. Obtaining a food product in liquid form

The following components of the food product (wt.%) Are introduced into the homogenizer:

Lipid and protein-amino acid fractions extract of crushed cysts, or nauplii, or adult crustacean Artemia salina in a ratio of 100: 1 0.1-1.0 vegetable oil the rest is up to 100%

Next, the resulting product is poured into containers.

Example 6. Obtaining a cosmetic product

In the manufacture of the drug, simple technological equipment is used, equipped with a reactor with a stirrer.

To prepare a cream, a gel base is loaded into a stirred reactor at a temperature of + (18-20) ° С and a biologically active additive is added with stirring according to the following formulation (wt.%):

lipid and protein-amino acid fractions extract of crushed cysts, or nauplii, or adult crustacean Artemia salina in the ratio of 500: 1 5.0-30.0 gel base the rest is up to 100%

The components are mixed for 20-30 minutes and the pH of the mixture is measured. If the pH of the mixture is below 7.2 (the gel has an acidic environment), then a phosphate-salt additive (Na ₂ HPO ₄ × 12H ₂ O) is added to the composition in an amount sufficient to maintain a pH of 7.2. Then the mixture is homogenized again in the reactor for 20-30 minutes, the finished cosmetic product is packaged in a container and stored at a temperature of + 7 ° C. The cold method of preparing the cream allows you to maximize the activity of its constituents.

Proteins in cysts, nauplii, and adult individuals of Artemia salina were determined by chromatography, and amino acids were determined by capillary electrophoresis.

Amino acid composition. 18 amino acids were found in the proteins of Artemia, the relative content of which changes during development (Table 1). The amount of free amino acids decreases in nauplii during the development period. However, amino acids such as proline, alanine, glycine, series, remain unchanged. The high content of essential amino acids is noteworthy: threonine, valine, methionine, isoleucine, leucine, phenylalanine, lysine and histidine. As you know, these amino acids have great biological value, since they are necessary for proper nutrition of organisms and the synthesis of proteins in them. At the same time, the highest content of essential amino acids was observed in nauplii as compared with the corresponding indices in cysts and adult individuals, which determines the advantages of Artemia larvae as an effective biological product.

Table 1 The content of total amino acids in Artemia at different stages of its development (%) Amino acid Cysts Nauplii Adults Cystine 1.6-1.8 1.0-3.4 2,3 Aspartic 10.6-22.8 4.7-14.2 9.3 Threonine 0.01-1.9 • 4.2-9.5 4.1 Series 5.8-8.6 3.9-13.1 4.8 Glutamine 15.2-19.5 5.9-14.8 14,4 Proline 4.9-5.9 4.2-11.2 5.3 Glycine 4.9-6.0 5.2-13.5 5.5 Alanya 13.7-16.3 3.6-24.0 7.0 Isoleucine 0.01-0.5 0.7-6.5 5.8 Leucine 0.6-1.4 1,5-9,5 8.2 Tyrosine 1.9-8.5 2.6-9.1 4,5 Histidine 0.8-1.8 0.8-4.2 1.8 Lysine 1.8-2.6 1.8-10.1 7.7 Arginine 3.2-6.3 1.8-9.9 6.6 Valine 0.02-2.0 2,3-8,3 5,6 Phenylalanine 0,0-0,2 1.7-8.9 4.7 Methionine - 1,5-3,2 2.7 Taurine 0.01-27.0 0,0-19,8 -

Squirrels. Artemia is high in protein. However, this indicator also varies depending on the stage of development of the crustacean, the race of Artemia and its food (Table 2). The share of soluble proteins is 29%, insoluble - 71%, the ratio of high molecular weight to low molecular weight - 43% and 57%, respectively.

table 2 The protein content in Artemia at different stages of its development (% to dry weight) Cysts Decapsulated Cysts Nauplii Adults 52.35 61.14 48.25 29.87

The protein content in Artemia cysts can vary in different years. In addition, for each stage of Artemia, specific proteins are characteristic, the identification of which is of great interest to elucidate their biological role and physiological activity.

The determination of lipids in the feedstock (cysts, or nauplii, or adults) was carried out by chromatographic method.

The lipid content in cysts, nauplii and adult Artemia individuals is presented in table 3. During the period of intensive growth, especially on the first day, the lipid concentration decreases in the range of 10-20%. The concentration of fat in Artemia cysts may vary depending on the time of collection.

Table 3 Artemia lipid content at different stages of development (%, to dry weight) Cysts Decapsulated Cysts Nauplii Adults 2.96 25.57 15.25 7.64 Table 4 The lipid composition of Artemia cysts (% of total lipids) Phospholipids Cholesterol Free fatty acids Triglycerides Esters of cholesterol 14.6 10.6 3.0 59.8 12.0

From the analysis of the lipid composition of Artemia cysts, it is seen (Table 4) that the main components of Artemia cysts are triglycerides.

Of particular importance are Artemia fatty acids, which are part of lipids and determine their properties. The total content of fatty acids increases during the development of crustacean: Artemia eggs contain 490-704 mg of fatty acids per 1 g of lipids, in nauplii this indicator increases to 602-854 mg / g.

Relatively large amounts of essential fatty acids are contained in cysts, nauplii, and adult crustaceans - C18: 2 (linoleic), C18: 3 (linolenic), and C20: 4 (arachidonic). Of the unsaturated fatty acids, Artemia is dominated by palmitic and stearic acids, of monounsaturated fatty acids - oleic and palmitoleic, of polyunsaturated ones - eicosapentaenoic, and in nauplii and cysts - linolenic.

High concentrations of essential fatty acids, reproductive, growth and adaptive hormones, vitamins, carotenoids, and essential amino acids determine the biological activity of the claimed product, which ensures metabolic recovery and promotes normal growth and development of the human and animal body. In addition, the dosed introduction of a protein-vitamin fraction (from 1000: 1 to 1: 1) into the lipid fraction, depending on the purpose of the final product, ensures a reduction in allergic reactions among consumers of this product.

Claims (1)

Dietary supplement based on cysts, or nauplii, or adult Artemia salina crayfish for external or internal use, characterized by the content of lipid and protein-amino acid fractions from the extract of crushed cysts, or nauplii, or adult Artemia salina crayfish in a ratio of 1000: 1 up to 1: 1.