Method for complex processing of fish raw material for obtaining hyaluronic acid and collagen
SUBSTANCE: skins of pond fish are flushed with cold flushing water during 10-15 minutes. They are crushed to the size of 2-3 mm. Water extraction is performed at the temperature of 40-45°C during 40-50 minutes at the ratio of crushed skins to water, which is equal to 1:1 at periodic mixing. Then, they are filtered; liquid fraction is dried in a spraying drier at the drier outlet product temperature of 60-65°C during 15-25 minutes so that hyaluronic acid is obtained. Solid fraction is subject to bleaching during 12 hours with hydrogen peroxide-salt solution that is prepared by mixing of 1 l of 3% hydrogen peroxide and 20 g of sodium chloride. Treatment of bleached solid fraction is performed with 1.0-1.2% solution of sodium hydroxide during 24 hours at the temperature of 20-25°C with further neutralisation of the obtained mixture with 3% boric acid solution. Treatment of swollen solid fraction is performed with Pancreatin ferment preparation solution taken in the quantity of 0.5-0.6% to the weight of solid fraction during 1.5-2.0 hours at the temperature of 37-40°C. Flushing of solid fraction is performed with cold flushing water for removal of Pancreatin residues so that collagen is obtained. The obtained collagen, depending on the purpose, is supplied for drying in drying chambers with forced air circulation at the temperature of 18-20°C during 12 hours and storage in dry ventilated rooms at the temperature of not higher than 20°C during 24 months or frozen to the temperature of minus 18 - minus 20°C and stored at the temperature of minus 18 - minus 20°C during 24 months. The liquid fraction dried in the spraying chamber is stored at the temperature of 0-4°C during 12 months or dissolved in physiological buffer solution.
EFFECT: improvement of the method.
2 dwg, 1 tbl, 1 ex
The invention relates to fish processing and cosmetic industries, and in particular to methods of processing the skins of fish to obtain hyaluronic acid and collagen.
The closest in technical essence and the achieved effect is a method of processing raw material containing collagen, providing preliminary freezing of raw materials, grinding, degreasing, alkali-salt processing, washing, neutralization, grinding the obtained collagen, dissolving it in the organic acid and homogenization [Patent No. 2139937. The method of processing raw material containing collagen / Aotion; Opotow; Vggrtusahela; Nambouwalu; Geantrai No. 2139937; Claimed 07.12.1998; Publ. 20.10.1999].
The disadvantages of the method are mnogovershinnoe and duration of the process; the use of more aggressive reagents; the necessity of dissolving the collagen in the organic acid.
The technical problem of the invention is to develop a method for integrated processing of fish raw materials for the production of hyaluronic acid and collagen, which allows to obtain hyaluronic acid and collagen for one cycle, to implement waste-free technology for the processing of pond fish through the best use of secondary products cutting, to expand the raw material base of the cosmetic industry, and is also to reduce the time and simplify the process of obtaining the target product.
To solve the technical problem of the invention a method for complex processing of fish raw materials for the production of hyaluronic acid and collagen, characterized in that for obtaining hyaluronic acid and collagen using the skins pond fish, which are obtained from fish processing plants skins pond fish washed with cold running water for 10-15 minutes, reduce them to a size of 2-3 mm, conduct water extraction at a temperature of 40-45°C for 40-50 min at a ratio of shredded skin:water is 1:1 with periodic stirring, filtered, the liquid fraction is dried in a spray drier at the temperature of the product leaving the dryer 60-65°C for 15-25 min with a production of hyaluronic acid, the solid fraction is subjected to bleaching peroxide-salt solution is taken so that all the solid fraction was covered peroxide-salt solution, which is prepared by mixing 3%hydrogen peroxide with sodium chloride, from the calculation: 1 l of 3%hydrogen peroxide is introduced 20 g of sodium chloride, the process is conducted within 12 hours, after this time the peroxide-salt solution is drained, then blanched solid fraction is subjected to swelling in 1,0-1,2%solution of a hydroxide sodium taken in 1:1 ratio to the mass of the solid fraction, 24 h at 20-25°C is followed by neutralization of the mixture 3%solution of boric acid, next, handle the swollen solids solution of enzyme preparation "Pancreatin" (for example, "Pancreatin" Irbit himfarmzavod, Russia, according to the manufacturer, the solid amorphous powder with enzyme activity: proteolytic - 200 FIP units (IU enzyme activity), amylolytic - 3500 FIP units, lytic - 4300 FIP units), taken in an amount of 0.5-0.6% by weight solids, for 1.5-2.0 hours at a temperature of 37-40°C, and the enzyme preparation is prepared by mixing with water in the ratio 1:3, then hold washing the solids with cold running water to remove residual "Pancreatin"derived collagen, depending on destination, sent for drying in the drying chamber with forced air circulation at a temperature of 18-20°C for 12 h and stored in a dry ventilated room at a temperature not exceeding 20°C for 24 months or frozen to a temperature of minus 18 to minus 20°C and stored at a temperature of minus 18 to minus 20°C for 24 months, dried in a spray drier, the liquid fraction is stored at a temperature of 0-4°C for 12 months or dissolved in a physiological buffer solution.
The technical result consists in a comprehensive receipt from the skins pond fish hyaluronic acid and collagen in one process cycle, EXT is of the functionality of secondary raw materials, the implementation of non-waste technology and deep processing of secondary products.
The method is as follows.
To obtain hyaluronic acid and collagen using the skins of the following species of fish: carp, carp, pike, carp, carp and others, the Proposed method can be used to produce hyaluronic acid and collagen from the skins of marine fish.
Adopted by the skin is subjected to washing with cold running water for 10-15 minutes to remove the mucus, dirt and retrieve globulin and albumen proteins. Next, the skin is crushed manual or automated way up to a size of 2-3 mm, because hyaluronic acid is soluble in warm water, then a water extraction of hyaluronic acid from the skin of pond fish. Extraction was carried out at a temperature of 40-45°C for 50 min with a ratio of the crushed skins:water is 1:1 with periodic stirring. The liquid is then filtered. The liquid fraction is used to produce GUK, and the solid fraction secrete collagen. The liquid fraction is dried in a spray drier at the temperature of the product leaving the dryer 60-65°C for 15-25 minutes Hyaluronic acid is stored in a dried form
at a temperature of 0-4°C for 12 months or dissolved in a physiological buffer solution.
Solid fraction of all Laut bleaching peroxide-salt solution, which is prepared by mixing 3%hydrogen peroxide and sodium chloride from the calculation: 1 l of 3%hydrogen peroxide is introduced 20 g of sodium chloride, peroxide-salt solution is taken in the same numbers, so that all the solid fraction was covered with it, the process is conducted within 12 hours, after this time the peroxide-salt solution is drained. Then bleached solid fraction is subjected to swelling in 1,0-1,2%sodium hydroxide solution taken in a 1:1 ratio to the mass of the solid fraction within 24 hours at 20-25°C, followed by neutralization of the mixture of 3%boric acid solution for 10 minutes Then swollen solid fraction is treated with a solution of enzyme preparation "Pancreatin" for the purpose of purification of collagen from the ballast globulin and albumen protein and fat components. The enzyme preparation contribute in the amount of 0.5 to 0.6% by weight solids, and the enzyme preparation is prepared by mixing with water in the ratio 1:3, the process is carried out for 1.5-2.0 hours at a temperature of 37-40°C, followed by washing with cold running water to remove residual "Pancreatin".
After washing in water, collagen or frozen to a temperature of minus 18 to minus 20°C or dried, and the drying is best conducted in the drying chambers with forced air circulation at a temperature which e 18-20°C during 12 hours
The dried collagen stored in dry, ventilated place at a temperature not higher than 20°C within 24 months, the frozen collagen stored at a temperature of minus 18 to minus 20°C for 24 months.
Method for integrated processing of fish raw materials for the production of hyaluronic acid and collagen is illustrated by the following example.
Example No. 1.
1 kg of skins carp subjected to washing in running cold water temperature of 18°C for 10 minutes to remove from the surface of the raw material mucus, dirt, extract ballast globulin and albumen proteins. Next skins carp crushed to a size of 2-3 mm manually, add 1 l of water, heat the mixture in thermostat to 40°C and incubated for 50 min at the same temperature and periodic stirring. After this time the liquid fraction is filtered off.
The liquid fraction is used to highlight the hyaluronic acid and the solid is used to obtain collagen.
The liquid fraction is dried in a spray drier at a temperature of product at the outlet of the apparatus 61°C for 15 minutes After drying, the product is a white amorphous powder, soluble in water. The output of biologic - 12% in terms of SV. Hyaluronic acid is stored in dried form at a temperature of 2°C for 12 months.
The solid fraction is subjected to bleaching,to do this, prepare a solution of 3%hydrogen peroxide and sodium chloride, taken from the calculation: 1 l of 3%hydrogen peroxide is introduced 20 g of sodium chloride and pour this solution on the solid fraction for 12 hours, so that all the solid fraction was covered peroxide-salt solution. After this time the peroxide-salt solution is drained.
Then bleached pulp for swelling pour 1 liter of a 1.0%aqueous solution of sodium hydroxide for 24 hours at 20°C. After swelling the mass of the solid fraction is 1.5 kg
Then hold the neutralization of sodium hydroxide 3%boric acid solution for 10 minutes
For purification of the solid fraction from the ballast globulin and albumen protein and fat components are treated with a solution of enzyme preparation "Pancreatin". The solid fraction add 3 l of water, heat the mixture to 37°C, contribute to 7.5 g of Pancreatin and maintain a 2.0 h at the same temperature and periodic stirring. After this time the solid fraction was washed with cold running water to remove residual enzyme preparation.
After rinsing in water the resulting collagen frozen at minus 18°C or dried, and the drying is best conducted in the drying chambers with forced circulation of air and at a temperature of 18°C for 12 hours
The dried collagen stored in dry ventilated by which edenia at a temperature not exceeding 20°C for 24 months.
As can be seen from table 1, the proposed method allows to obtain hyaluronic acid and collagen in one process cycle, hyaluronic acid, after drying, is a white amorphous powder, soluble in water, the yield of hyaluronic acid is 10-12% in terms of MW, the output of collagen - 45-50% by weight of the crushed skins. In the process of water extraction a significant impact on the yield of hyaluronic acid has a temperature. When the temperature reaches 50°C maximum observed output of the biopolymer, the use of 3%hydrogen peroxide leads to a significant increase in interstitial pressure in the connective tissue, pushing its structural elements, causes the formation of multiple ties radical type between the active groups of the side chains of collagen molecules and allows you to achieve in the future better absorption of sodium hydroxide. Analysis of experimental data shows that upon receipt of collagen after 3 h after the beginning of the peroxide-alkaline hydrolysis the degree of swelling of the skins of fish is about 50%. The maximum value of the degree of swelling of the sample is observed after 24 h after the beginning of the hydrolysis. The mechanism of action of sodium hydroxide is in violation and the weakening of some of the hydrogen bridges, partial rupture of protein-carbohydrate member is, surrounding the bundles of collagen fibrils, the impact of sodium hydroxide and hydrogen peroxide promotes better penetration of alkali in the structure of collagen fibrils.
The impact of "Pancreatin" leads to a weakening and partial rupture of relations between protein, protein-lipid complexes, which leads to the extraction from the skins of mucopolysaccharides, leaching protein formations structureless substance of the dermis.
In the method for integrated processing of fish raw materials for the production of hyaluronic acid and collagen possible use of the enzyme preparation "Pancreatin" different manufacturers (Bovines, Russia; Mikos, Russia; OOO Indukern-Rus, Russia; Samson-Med, Russia; Hubei Maxpharm industries Co., China; Biozym Gesellshaft fur Enzymtechnologie GmbH, Germany, and others)that have similar enzymatic activity with the enzyme preparation "Pancreatin" Irbit himfarmzavod, Russia.
Drying of collagen best conducted in the drying chambers with forced air circulation at a temperature of 18-20°C for 12 H. the Advantage of this method is that the dried collagen in appearance corresponds to the given type of product, drying does not require additional power costs.
A water extraction for isolation of hyaluronic acid at a temperature above 50°C does not lead to substantially the mu-change output GUK, but creates conditions for the development of denaturation and coagulation processes, reducing the purity and quality of the drug. Temperatures below 50°C reduces the rate of extraction, and therefore slows down the whole technological cycle.
When using hydrogen peroxide at a concentration below 3%, does not achieve the desired effect, and the use of hydrogen peroxide in concentrations above 3%, results in excessive oxidant.
The proposed method allows to obtain hyaluronic acid and collagen for one cycle, to implement waste-free technology for the processing of secondary products cutting pond fish, rational use of Malostranske raw materials of the fishing industry to reduce the duration of the technological process, to use the skins of all fish species, including marine.
|Criteria comparison||The placeholder||A new technical solution|
|The reception of the skin of pond fish||No data||Skins take lots|
|Flushing||Ethyl alcohol with the purpose of bleeding cocks ' combs (raw material:ethyl alcohol 1:2)||Rinse with cold running water (τ=10-15 min, t=18-20°C)|
|Grinding||Grinding skins to a size of 2-3 mm||Grinding skins to a size of 2-3 mm|
|Extraction||A solution of the enzyme preparation "Collagenase" content 0,035-0,040% by weight of raw materials for 45-50 minutes at a temperature of 45-50°C and a pH of 6.8 to 7.0.||Water (τ=40-50 min, t=40-45°C)|
|Deposition GUK||The deposition of 96%ethyl alcohol (liquid:ethanol 1:3)||Not required|
|Drying||Freeze-drying||Drying in a spray dryer (τ=15-25 min, t=60-65°C)|
|Whitening||No data||3%hydrogen peroxide solution, 2% sodium chloride, τ=12 h|
|Swelling||Molochnosokovoj bath, containing 4-5% NaOH and 6-7% Na2SO4, LCD=3.||1,0-1,2%solution of sodium hydroxide, τ=24 h, t=20-25°C|
|Neutralization||90 minutes at a temperature of 18-20°C ammonium sulfate in an amount of 5% by weight of the feedstock under stirring||3%boric acid solution, τ=10 min|
|Enzymatic processing||No data||Processing enzyme preparation "Pancreatin" (t=37-40°C; τ=1.5 to 2.0 hours; dosage of 0.5-0.6% by weight of raw material, LCD=1-3)|
|The total time of the process (without drying or freezing)|
|There is insufficient data||Less than 2 days|
Method for integrated processing of fish raw materials for the production of hyaluronic acid and collagen, which wash with cold running water for 10-15 minutes skins pond fish, grinding skins to a size of 2-3 mm, water extraction at a temperature of 40-45°C for 40-50 min at a ratio of crushed raw material:water 1:1 with periodic stirring, filtering, drying of liquid fraction in the spray dryer at a temperature of the product leaving Sushil is 60-65°C for 15-25 min with a production of hyaluronic acid, the solid fraction is subjected to bleaching within 12 h of peroxide-salt solution prepared by mixing 1 l of 3%hydrogen peroxide with 20 g of sodium chloride, processing bleached solid fraction 1,0-1,2%sodium hydroxide solution for 24 h at 20-25°C, followed by neutralization of the mixture 3%solution of boric acid, solid fraction enzyme preparation "Pancreatin", taken in an amount of 0.5-0.6% by weight solids, for 1.5-2.0 hours at a temperature of 37-40°C, washing the solid faction cold running water to remove residual "Pancreatin" with the receipt of collagen derived collagen, depending on destination, sent for drying in the drying chamber with forced air circulation at a temperature of 18-20°C for 12 hours or freeze up to a temperature of minus 18 to minus 20°C.
SUBSTANCE: strain is built by transformation of strain-recipient Escherichia coli BL21 (DE3) with plasmid DNA containing the coding area of gene aehR of hydrolase of alpha-amino acids esters Xanthomonas rubrilineans All-Russian collection of industrial microorganisms B-9915 under control of promoter T7 and gene of stability to kanamycin Kan of hydrolase of alpha amino acids esters from Xanthomonas rubrilineans All-Russian collection of industrial microorganisms B-9915. The proposed synthesis method of hydrolase of alpha-amino acids esters is implemented by cultivation of strain-producer Escherichia coli All-Russian collection of industrial microorganisms B-11246 with addition to the composition of environment of kanamycin and isopropyl-"в"-D-tiogallactoside as an expression inducer of the obtained product.
EFFECT: high biosynthesis level of target product, which can be used for development of methods for obtaining biocatalysts of synthesis processes of aminopenicillins and aminocephalosporins.
2 cl, 3 dwg, 1 tbl, 5 ex
SUBSTANCE: fermentation of nutritional medium is carried out based on corn starch hydrolysates by actinomycete. Then ultrafiltration of the obtained filtrate of the culture liquid is carried out with an activity of 4250±250 IU/cm3 through membranes with cut-off of 5 kDa. The filtrate obtained is concentrated under vacuum of 0.082 mPa at a temperature of 25-30°C. It is clarified at the temperature of 30-40°C. Its ultrafiltration is carried out again through the membrane with cut-off of 1 kDa. The obtained glycosidase inhibitor is dried and ground.
EFFECT: invention enables to obtain the glycosidase inhibitor with increased activity.
1 tbl, 4 ex
SUBSTANCE: isolated recombinant adenosine deaminase is described, which comprises polypeptide SEQ ID NO: 1 or a version polypeptide SEQ ID NO: 1 of isolated recombinant adenosine deaminase, where the version polypeptide SEQ ID NO: 1 comprises one or more amino acid substitutions selected from the group consisting of: Gin instead Lysl98; Ala instead Thr245; and Arg instead of Gly351, and DNA encoding it. The conjugate of polyalkylene oxide with the said adenosine deaminase for treatment of adenosine deaminase-mediated diseases is presented where adenosine deaminase comprises from 11 to 17 chains of polyalkylene oxide with a molecular weight of 5 kDa for ADA protein. The methods of purification of the recombinant adenosine deaminase are proposed, including protein purification using ion exchange chromatography, or protein purification using hydrophobic interaction chromatography. Also the preparations of recombinant adenosine deaminase produced by these methods are provided.
EFFECT: invention enables to obtain recombinant adenosine deaminase, having increased stability.
14 cl, 1 tbl, 10 ex
SUBSTANCE: bacteria strain Planomicrobium koreense 78k, is extracted from soils and provides for production of site-specific endonuclease PkrI, which recognises and splits both chains of nucleotide sequence of DNA, which contains three or four C5-methylcytosine bases in recognition site 5'-GCNAGC-3' with formation of a single-nucleotide 3'-protruding end. The strain is deposited in the Russian National Collection of Industrial Microorganisms FGUP GosNIIgenetika under the number B-10627. The provided strain may be used for extraction of the new site-specific endonuclease PkrI, which may be applied in detection and splitting of methylated DNA sections.
EFFECT: production of site-specific endonuclease PkrI.
3 dwg, 3 ex
SUBSTANCE: invention represents a bacteria strain Microbacterium testaceum 17B, deposited under the number RNCIM B-10628 in the Russian National Collection of Industrial Microorganisms FGUP GosNIIgenetika, being the producent of the site-specific endonuclease MteI. The invention makes it possible to produce the site-specific endonuclease, which recognises and splits both chains of the nucleotide DNA sequence 5'-G(m5C)G(m5C)∧NG(m5C)G(m5C)-3'/3'-(m5C)G(m5C)GN∧(m5C)G(m5C)G-5', where m5C-5-methylcytosine, with production of the single-nucleotide 5'-protruding end, and does not split the methylated DNA sequence 5'-G(m5C)NG(m5C)-3'/3'-(m5C)GN(m5C)G-5'.
EFFECT: method improvement.
2 dwg, 1 tbl, 3 ex
SUBSTANCE: supernatant containing a recombinant protein with an amino acid sequence of staphylokinase (RPSF) is defrozen. The RPSF is recovered and chromatographically purified on CM-sepharose. The RPSF is chromatographically purified on DEAE-sepharose. Then the RPSF is transferred into 0.3% NaCl solution. It is followed by sterilisation, packaging, lyophilisation and marking of the substance. 20 litres of water for injections are introduced in a stirred reactor 160 l, then 150 g of the substance and 10 l of water for injections are added; the stirrer is actuated (stirring rate is 80 rpm); the stirring procedure is carried out for 15 min; thereafter the stirrer is stopped; the substance is sampled; the pH value of the prepared solution is recorded; pH is 5.5 to 7.5. It is followed by sterilisation filtration of the solution, filling and pre-corking of bottles. Then lyophilisation and final corking are performed. What is offered is the application of the preparation Fortelysin for treating myocardial infraction by the intravenous introduction in dose 15 mg (2235000 IU) with the content of a bottle 5 mg (745000 IU) before the introduction of the preparation is dissolved in 0.9% sodium chloride 5 ml; the preparation is introduced in two boluses 10 mg (1490000 IU) simultaneously and in 30 minutes 5 mg (745000 IU) or at first 10 mg (1490000 IU) in boluses, and the rest 5 mg (745000 IU) in intravenous infusions in 0.9% sodium chloride 50 ml for 30 min.
EFFECT: invention extends the range of drug preparations with fibrinolytic properties.
SUBSTANCE: disclosed is a plasmid which determines synthesis of alkaline phosphatase CmAP, containing a NcoI/SacI-fragment of plasmid pET-40b(+) (Novagen) and a DNA fragment of 1530 base pairs, which contains a chimeric gene consisting of a CmAP gene structural part which is adapted at the N-end for expression in E.coli cells, and a nucleotide which codes a specific sequence for TEV protease. Described is an E.coli Rosetta(DE3) strain which is transformed by said plasmid, and is a producer of chimeric protein, which contains an amino acid sequence of recombinant alkaline phosphatase CmAP. Disclosed is a method of obtaining recombinant alkaline phosphatase CmAP, comprising steps for: incubating said producer strain in a liquid broth LB for 12 hours at 20°C, depositing bacterial cells by centrifuging, disintegration of the cell suspension in a buffer, centrifuging the extract, chromatography of the supernatant fluid on a column with a metal affinity resin, elution of the protein, concentrating active fractions by ultrafiltration, incubation with protease TEV, concentrating the protein solution and extracting the end product by gel filtration.
EFFECT: improved method.
3 cl, 1 dwg, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to veterinary, agriculture, medicine, biotechnology and microbiological industry and can be applied for protection from infections, induced by DNA - and RNA- containing viruses. Antiviral combined medication of contact action based on "Betadine", represents mixture of medication "Betadine" with solution of bacterial endonuclease - product of gen nuc A (synonym nuc), in any ratio, and content of povidone-iodine in diluted medication "Betadine" constitutes 0.005 - 0.000005%, and pH of solution of bacterial endonuclease in buffer or salt solution, which contains magnesium cations, constitutes 7-9.
EFFECT: medication has simultaneously reduced irritating effect and high virulicidal activity.
SUBSTANCE: Trichoderma aureoviride Rifai fungus strain is recovered from sea ground collected at depth 25 m in the South China Sea. It is deposited in the Russian National Collection of Microorganisms of G.K.Skryabin Institute of Microbial Biochemistry and Physiology of the Russian Academy of Sciences, registration No. VKM F-4115D. The strain is cultivated with using a nutrient medium not containing sugars. Total yield pf 1,3-β-D-glucanase is 13.5-16 units/mg of protein.
EFFECT: strain is characterised by high yield of 1,3-β-D-glucanase.
1 tbl, 2 ex
SUBSTANCE: claimed is method of determining DNA-hydrolysing activity of molecules and device for its realisation. Method includes adsorption of reaction mixture, which contains DNA and tested substances, on working electrode, modified with carbon nanomaterials. After that, registered and compared are values of DNA oxidation in composition of reaction mixture before standing reaction mixture (control) and after standing reaction mixture (experiment). Increase of DNA oxidation signal testifies to presence of DNA-hydrolysing activity in tested molecule.
EFFECT: invention makes it possible to simplify and reduce terms of determining DNA-hydrolysing activity of molecules, increase selectivity and sensitivity of the test reducing its cost.
8 cl, 6 dwg, 2 ex
SUBSTANCE: production method of glucan-chitosan complex from yeast biomass of brewing waste involves mechanical and ultrasonic treatment of yeast biomass, destruction of proteins by treatment of the obtained suspension using alkali reagents with further extraction of a target product. As biomass, Saccharomyces living yeast is used. First, yeast is frozen to -15°C during 24 hours. After mechanical destruction, biomass is treated for 15 minutes at 20°C in an ultrasonic bath with frequency of an emitter of 35 kHz and voltage of 285 W. Biomass is acidified with chlorhydric acid till pH=5.5 and treated with ferment preparation in the amount of one pellet containing lipase - 3500 units of Ph.Eur., amylase - 4200 units of Ph.Eur. and protease - 250 units of Ph.Eur. per kilogramme of biomass in terms of dry substance; then, lipid components of yeast are removed. Fermentation is performed at t=20-29°C during 30-60 minutes. Destruction of proteins is performed at 55°C by means of a water bath during 60 minutes by treatment using 4% water solution of caustic soda at the ratio of yeast biomass and alkali, which is equal to 1:4. The medium is neutralised and hydrosol of glucan-chitosan complex is deposited by centrifugation during 10 minutes. The deposit is dried at t=55°C during 48 hours.
EFFECT: invention allows improving the quality of the obtained complex and its biological activity.
SUBSTANCE: invention relates to production of hydroxyalkyl derivatives of polysaccharides. The method of producing 2,3-dihydroxypropyl chitosan involves reacting chitosan with glycidol in the presence of hydrochloric acid with ratio glycidol:chitosan:hydrochloric acid = (2-6):1:1, at room temperature until a gel forms. The mixture is then heated at 55-65°C for 12-14 hours and the reaction mass is treated with water. The mixture is then deposited, subjected to hot extraction with water-soluble alcohols or ketones and dried.
EFFECT: invention simplifies the method of production and output of the end product and improves sorption properties of the compound.
1 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: there are presented: using benzophenanthridine alkaloid salts for preparing therapeutic agents for treating tumours, wherein the alkaloid salt is found in the form luteic, phosphatidic or hyaluronic acid, the benzophenanthridine alkaloid salt with phosphatidic acid or hyaluronic acid, and a based pharmaceutical composition for treating tumours.
EFFECT: what is shown is cytotoxic activity of the sanguinarine salts according to the invention at least twice increased in all studied tumour cell lines in relation to the chloride salt; it is suggested to be caused by higher absorption by the tumour cells.
12 cl, 8 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to a recovered imidised biologically compatible polymer functionalised by an imide group. The above polymer is selected from the group consisting of polyethylene oxide, partially or completely hydrolysed by polyvinyl alcohol, polyvinylpyrrolidone, polyethyloxazoline, polyoxypropylene oxide block copolymers (poloxamers and meroxapol), polyethylene oxide and poloxamine copolymer, carboxymethyl cellulose and hydroxyalkylated cellulose, polypeptides, polysaccharides, carbohydrates, polysaccharose, hyaluronic acid, dextran, heparin sulphate, keratan sulphate, chondroitin sulphate, heparin, alginate, gelatin, collagen, albumin, ovalbumin, complex polyphosphoesters, polylactides, polyglycolides, polycaprolactones, polyamides, polyurethanes, polyesteramides, polyorthoesters, polydioxanones, polyacetals, polyketals, polycarbonates, polyorthocarbonates, polyphosphazenes, polyhydroxybutyrates, polyhydroxyvalerates, polyalkylene oxalates, polyalkylene succinates, polymaleic acids, polyamino acids, polyvinyl alcohol, polyvinylpyrrolidone, polyhydroxy cellulose, chitin, chitosan, and copolymers, ternary copolymers, or combinations or mixtures of the aforementioned materials. Also, the invention refers to a composition for a tissue adhesive, a medical device and a pharmaceutical composition.
EFFECT: invention represents additionally modified or functionalised imidised polymers.
9 cl, 2 ex, 20 dwg
SUBSTANCE: fish roe is homogenised. Fish roe hydrolysis is carried out with a ferment preparation "Collagenase" in presence of an inhibitor for 10-12 hours. Chitosan is added to the produced hydrolysate at the ratio of 0.5-1.0:1.0. Components are mixed.
EFFECT: invention makes it possible to accelerate process of chitosan-nucleic complex production.
1 dwg, 1 tbl, 3 ex
SUBSTANCE: method includes depolymerisation of a high-molecular chitosan with hydrogen peroxide. The process of chitosan depolymerisation is carried out in a double-phase system. The solid phase is activated chitosan with Mav = 450-650 kDa and the average particle size of 0.05-0.20 mm. The liquid phase is a water solution of H2O2 with concentration of H2O2 in a reaction system equal to 1-7%. The reaction is carried out for 120-180 minutes at 70°C. Then phase separation of produced chitosan homologs is carried out by means of filtration via paper or textile surface of the produced reaction mixture. The produced filtrate contains water-soluble chitosan oligomers.
EFFECT: invention makes it possible to quantitatively control extent of conversion of an initial high-molecular chitosam into oligomer and low-molecular structures of its homologs.
1 tbl, 1 ex
SUBSTANCE: invention relates to a method of purifying chondroitin sulphate and can be used in food and cosmetic industry and in medicine. The method involves electrochemical deposition to obtain a hydrogel of chondroitin sulphate, stabilisation, removal from the electrode, washing and drying. The chondroitin sulphate is dissolved in a 0.01-0.1 n alkali solution in ratio of 1:50-1:200 and deposited in an alkaline medium with constant cooling and stirring. The solution is stirred at a rate of 10-20 rpm. Current density is equal to 1-10 A/m2. Voltage is preferably not lower than 2.7 V. The hydrogel of chondroitin sulphate is stabilised in a 0.05-0.5 n HCl solution.
EFFECT: invention enables to obtain chondroitin sulphate with high weight ratio of the basic substance and increases output of the end product.
5 cl, 1 ex
SUBSTANCE: present invention relates to a method of producing a N,S-cyclo-containing chitosan derivative. Described is a method of producing a chitosan-based N,S-cyclo-containing polymer (I) which contains in the macrochain 1-oxa-6-thia-4,8-diazocycloundecane fragments: I, by reacting chitosan with formaldehyde and a S-containing compound, characterised by that the S-containing compound used is hydrogen sulphide, the formaldehyde solution is pre-saturated with H2S and the reaction is carried out with molar ratio chitosan: formaldehyde: hydrogen sulphide of 1:2:1, at temperature of 0-60°C in a chloride medium for 24 hours.
EFFECT: obtaining modified chitosan which exhibits properties of a highly efficient heavy metal sorbent for waste water treatment, an extractant for separating rare, noble and precious metals and a complexing agent for biological molecules.
1 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biochemistry. What is presented is a conjugate of hyaluronic acid and novocaine of a structure as defined in the patent claim containing 20-50% residues of novocaine.
EFFECT: conjugate is water-soluble; it possess the amphoteric properties and contains no side O-acylisourea.
3 tbl, 3 ex
SUBSTANCE: disclosed are versions of a method of producing cross-linked polysaccharides, involving reaction of at least one polysaccharide selected from amino-polysaccharide, amino-functionalised polysaccharide containing one or more amino groups which can be cross-linked by reducing sugar, and combinations thereof, with at least one reducing sugar. The invention also discloses polysaccharides obtained using the disclosed method, a method of producing cross-linked matrices based on polysaccharides and matrices obtained using this method. The obtained matrices may include polysaccharide matrices and composite cross-linked matrices, including polysaccharides cross-linked with proteins and/or polypeptides.
EFFECT: obtained polysaccharides have satisfactory resistance to enzymatic degradation coupled with rheological properties of the preparation for injection, obtained matrices exhibit various physical, chemical and biological properties.
29 cl, 12 dwg, 6 tbl, 11 ex
SUBSTANCE: peptides consist of four amino-acid residues that are used for stimulation of collagen production with fibroblast.
EFFECT: invention allows effective stimulation of collagenoses in fibroblast cells.
11 cl, 3 dwg, 7 tbl, 6 ex