Method of producing chondroitin sulphate from sea hydrobiont tissue. RU patent 2458134.
| Synthetic oligonucleotide primers and method for detecting streptococcus thermophilus in starter cultures in hard cheese production / 2455363 There are offered synthetic oligonucleotide primers and method for detecting Streptococcus thermophilus in starter cultures in hard cheese production. The presented method involves conducting the PCR. If having found the DNA fragment of size of 655 b.p., the presence of Streptococcus thermophilus in the analysed biomaterial is stated. |
 Recombinant dna and method of obtaining l-lysine / 2451748Invention relates to biotechnology. The invention relates to a method of obtaining L-lysine using Escherichia coli bacteria containing versions of the dapA gene of B. subtilis bacteria. |
 Method of producing tagatose using soya oligosaccharide / 2451688Invention relates to a method of producing tagatose. The method involves the following steps: a) extraction of soya oligosaccharide from by-products of soyabean whey; b) hydrolysis of soya oligosaccharide using α-galactosidase which does not contain invertase; c) isomerisation of the hydrolysed sugar solution using arabinose isomerase to obtain an isomerised product containing mixed sugar containing saccharose, galactose and tagatose as the basic components; and d) separation of the isomerised solution via chromatography. |
| Biocatalyst, method of preparing said biocatalyst and method of obtaining invert syrup using said catalyst / 2451546 Invention is meant for use in food industry and specifically relates to the technology of obtaining syrups containing glucose and fructose, used in confectionary and baking industry to make confectionary products and candy. Disclosed is a biocatalyst for producing invert syrup, a method of producing said biocatalyst and a method of producing invert syrup - an inversion of saccharose using the prepared biocatalysts. The biocatalyst for producing invert syrup contains the following in wt % on dry substances: yeast autolysate as enzyme-active biomass 30-50, nanocarbon component 5-15 and filler consisting of silicon dioxide - up to 100. A nanocarbon component with a nanofibre structure or a carbon nanotube structure or a nanodiamond structure or a bulbous nanocarbon structure is used. The method of obtaining invert syrup is realised in a reactor with a fixed bed of the biocatalyst described above at temperature not higher than 50°C. |
 Fermentative production of organic compounds / 2451081Starch-containing raw materials (cereals) c are milled to produce a milled product including solid starch-free components in an amount of no less than 20 wt % One prepares a water-based suspension of the milling product to ultimately obtain flour pulp with dry weight content equal to at least 45 wt %. The pulp is heated in a jet boiler by way of water steam delivery to a temperature exceeding that of starch gelatination. One performs flour pulp hydrolysis by way of dilution and subsequent saccharification in the presence of α-amylase enzyme in a quantity of 0.002 - 3.0 wt % of the total quantity of the starch-containing raw material used to produce water medium M. Water medium M is added to the fermentative medium wherewith one cultivates a microorganism capable of the organic compound hyperexpression for fermentative broth production. |
| Fermentative production of organic compounds / 2451081 Starch-containing raw materials (cereals) c are milled to produce a milled product including solid starch-free components in an amount of no less than 20 wt % One prepares a water-based suspension of the milling product to ultimately obtain flour pulp with dry weight content equal to at least 45 wt %. The pulp is heated in a jet boiler by way of water steam delivery to a temperature exceeding that of starch gelatination. One performs flour pulp hydrolysis by way of dilution and subsequent saccharification in the presence of α-amylase enzyme in a quantity of 0.002 - 3.0 wt % of the total quantity of the starch-containing raw material used to produce water medium M. Water medium M is added to the fermentative medium wherewith one cultivates a microorganism capable of the organic compound hyperexpression for fermentative broth production. |
| Fermentative production of organic compounds / 2451081 Starch-containing raw materials (cereals) c are milled to produce a milled product including solid starch-free components in an amount of no less than 20 wt % One prepares a water-based suspension of the milling product to ultimately obtain flour pulp with dry weight content equal to at least 45 wt %. The pulp is heated in a jet boiler by way of water steam delivery to a temperature exceeding that of starch gelatination. One performs flour pulp hydrolysis by way of dilution and subsequent saccharification in the presence of α-amylase enzyme in a quantity of 0.002 - 3.0 wt % of the total quantity of the starch-containing raw material used to produce water medium M. Water medium M is added to the fermentative medium wherewith one cultivates a microorganism capable of the organic compound hyperexpression for fermentative broth production. |
| Fermentative production of organic compounds / 2451081 Starch-containing raw materials (cereals) c are milled to produce a milled product including solid starch-free components in an amount of no less than 20 wt % One prepares a water-based suspension of the milling product to ultimately obtain flour pulp with dry weight content equal to at least 45 wt %. The pulp is heated in a jet boiler by way of water steam delivery to a temperature exceeding that of starch gelatination. One performs flour pulp hydrolysis by way of dilution and subsequent saccharification in the presence of α-amylase enzyme in a quantity of 0.002 - 3.0 wt % of the total quantity of the starch-containing raw material used to produce water medium M. Water medium M is added to the fermentative medium wherewith one cultivates a microorganism capable of the organic compound hyperexpression for fermentative broth production. |
| Method for preparing proteinase ulp275 / 2451076 Strain Escherichia coli Russian National Collection of Industrial Microorganisms B-10996 is cultivated on a nutrient conditioned medium containing: peptone 2-4 wt %, yeast extract 1 - 2 wt % and water the rest in the presence of a selective antibiotic to stationary growth phase at temperature 20 °C to 30°C. The prepared is dissolved in a fresh portion of the same medium in the ratio 1:1 to 1:3. Simultaneously an inducer is added, and the cultivation is continued to for at least 4 hours. Synthesised proteinase is purified. |
 Enzymatic demethylation of flavonoids / 2449018Invention relates to biotechnology. The method of producing 8-prenylnaringenin in vitro involves: a) providing a first composition containing a culture of Eubacterium limosum or Peptostreptococcus productus bacterial cells, having 5-alkoxyflavonoid-dealkylating activity, and b) bringing a second composition containing 5-alkoxyflavonoids into contact with said first composition to facilitate dealkylation of said 5-alkoxyflavonoids with said bacterial cell culture. Disclosed is a combination for obtaining 8-prenylnaringenin in vitro and use of the culture of Eubacterium limosum or Peptostreptococcus productus bacterial cells, having 5-alkoxyflavonoid-dealkylating activity to obtain 8-prenylnaringenin in vitro. The method of enhancing 5-alkoxyflavonoid-dealkylating activity of the culture of Eubacterium limosum or Peptostreptococcus productus bacterial cells involves a step for inoculating said bacterial cell culture on a medium containing 5-alkoxyflavonoids, and a step for selecting a colony with the highest 5-alkoxyflavonoid-dealkylating activity. The Eubacterium limosum LMG P-23546 strain has 5-alkoxyflavonoid-dealkylating activity and is capable for converting isoxanthohumol to 8-prenylnaringenin. |
 Cross-linked hyaluronic acid and production method thereof / 2456299Method involves activation of hyaluronic acid using a cross-linking agent and an auxiliary cross-linking agent. The activated hyaluronic acid then reacts with a nucleophilic cross-linking agent. The pH of the reaction medium ranges from 8 to 12. The nucleophilic cross-linking agent contains at least 50 wt % oligopeptide or polypeptide. Further, pH of the reaction medium is regulated to 5-7 and cross-linked hyaluronic acid is precipitated in the organic solvent. The invention also relates to use of the cross-linked hyaluronic acid obtained using this method in plastic surgery to make implants and to a hedrogel containing said cross-linked hyaluronic acid in a buffer aqueous solvent. |
 Method of determining antibacterial activity of chitosan / 2450022Disclosed is a method of determining antibacterial properties of chitosan by estimating its minimum bacteriostatic and/or bactericidal concentration. Complex buffer solutions based on three organic acids MES, ACES and TES with different pH values are prepared. The ready buffer solutions are poured into a vessel. Double dilutions of chitosan are then prepared in vessels with the buffer solutions. Aliquots of a bacterial suspension in a fluid medium are added to the chitosan solutions in the buffer. The solutions are incubated for 24 hours at temperature which is optimum for bacterial growth. The minimum bacteriostatic and/or minimum bactericidal concentration of chitosan is then determined after incubation by determining growth of the culture or a drop in the number of living cells, respectively. |
| Method for preparing boron-containing hyaluronic acid / 2445978 Invention refers to a method for preparing sodium salt of hyaluronic acid modified by boron compounds with no fluid medium added. The method consists in the fact that powdered sodium salt of hyaluronic acid together with a modifying agent and mixed modifying agents is pre-homogenised in a mixer at temperature ranging within 20° to 50°C; thereafter the prepared homogenous powder mixture is simultaneously exposed to pressure and shearing deformation in a mechanochemical reactor at temperature ranging within 20° to 50°C and pressure 5-1000 MPa. |
 Method for producing chitosan oligomers / 2445101Invention refers to medicine, more specifically to producing chitosan oligomers possessing biological activity and applicable in food industry and medicine. In a method for producing chitosan oligomers, a chitosan solution is taken in the concentration of 0.025-0.075% (weight/volume) and exposed to low-frequency ultrasound of the intensity of 92-460 Wt/cm2 for 5-30 minutes. |
| Method of producing d(+) glucosamine hydrochloride / 2440362 Method involves preliminary acetylation of chitin with acetic anhydride, washing and drying the acetylated chitin in order to reduce degree of deacetylation thereof and, as a result, increase output of the desired product - D(+)-glucosamine hydrochloride when obtaining said product through hydrolysis of acetylated chitin with concentrated hydrochloric acid while heating, followed by evaporation, crystallisation, separation, washing and drying the desired product. |
 Chitosan chromate, synthesis method thereof and energy-intensive composition containing said chitosan chromate / 2439081Method of producing chitosan chromate involves reaction of soluble chitosan salts with metal chromates in ratio of 2 moles of the chitosan cation to 1 mole of chromate anion or with metal bichromates in ratio of 4 moles of the chitosan cation to 1 mole of the bichromate anion. The solid chitosan chromate residue formed is then separated and dried at temperature not higher than 150°C. The invention discloses an energy-intensive composition based on chitosan dodecahydro-closo-dodecaborate containing an effective amount of chitosan chromate. The quantitative ratio in the energy-intensive composition is by the required combustion mode: the higher the content of chitosan chromate, the higher the activity of the composition. |
 Method of determining degree of deacetylation of chitosan / 2436798Method involves taking a certain weighed amount of chitosanium chromate which is first purified from extraneous impurities and reduced to constant weight. The weighed amount is then turned into a stable weighted form through thermal treatment on air at temperature 800-900°C to form chromium oxide Cr2O3. The weight of the formed chromium oxide is then determined. Content of chromic acid in the initial weighed amount of chitosanium chromate is then calculated from the weight of chromium oxide. The degree of deacetylation of chitosan is calculated using defined formulae. |
 Method of extracting and stabilising low-molecular aminoglycans from eggshell wastes / 2435786Invention relates to a method of extracting and stabilising ultra low-molecular aminoglycans from eggshell wastes. Aminoglycan extract is used to produce cosmetic creams with skin moisturising and anti-wrinkle properties. The method of extracting low-molecular aminoglycan compound of formula I from a natural source of eggshell wastes, which consists of alternating glucuronic acid and N-acetylglucosamine units, where M can be one or more of Na, Ca, K, Mg; and n is a whole number from 20 to 40, involves the following steps: (a) preparing eggshell wastes for extraction of embryonic low-molecular aminoglycan compound of formula I using a polar organic solvent in water, (b) extracting low-molecular aminoglycan compound of formula I in form of a water-soluble salt, for which the eggshell from step (a) is vigorously shaken with aqueous polar salt solution at 10°C - 35°C for 6-12 hours, then filtered or centrifuged in order to collect an aqueous layer containing a dissolved aminoglycan compound of formula I; (c) extracting a purified low-molecular aminoglycan compound of formula I by forming a gel from an aqueous mixture of salts using a polar organic solvent, for which the solution from step (b) is successively and step-by-step mixed with an organic solvent mixed with water while gently stirring and then cooled to maintain temperature from 20°C to 25°C, and the formed gel is left for 2-24 hours for complete precipitation, then filtered or centrifuged in order to extract a semidry aminoglycan compound of formula I; (d) the extracted aminoglycan compound of formula I from step (c) is stabilised via gradual addition of organic oils to the semidry gel to form aminoglycan compound of formula I. In order to prepare a composition having anti-wrinkle properties, at least one pharmaceutically acceptable filler is added to the stabilised aminoglycan compound of formula I obtained at step (d). |
 Method and apparatus for treating fermented wastes and producing chitin containing biomass / 2430929Method involves feeding wastes to be treated into artificial containers, biotreatement, tapping the filtrate and removing the obtained biomass. Biotreatement is carried out by culturing hoverfly larvae from the freshly laid eggs phase to the pupation phase in the fermented wastes to be treated, placed in artificial meshed containers the bottom and walls of which are covered with filter cloth. The apparatus has artificial containers, devices for feeding the wastes to be treated, outputting the filtrate and collecting the biomass. The artificial containers have a meshed bottom and walls covered with filter cloth. |
| Method of producing nanoparticles of low-molecular chitosan / 2428432 Method of producing nanoparticles of low-molecular chitosan involves preparing a solution of pre-purified low-molecular chitosan in filtered 1-2 wt % aqueous acetic acid, adding solutions of hydroxides of alkali metals or ammonia for 2 hours, dispersing the system using a mechanical mixer at a rate of 200-300 rpm at temperature 20°C to pH 6.9-7.0. Further, the dispersion is centrifuged at 10000 rpm. The obtained solid residue is redispersed in bidistillate while mechanically mixing at a rate of 200-300 rpm at temperature 20°C. |
 Method of treating degeneratively-dystrophic diseases of musculoskeletal system and posttraumatic pain syndromes / 2433844Invention relates to medicine, namely to traumatology and orthopedics and can be applied in complex treatment of posttraumatic, dystrophic, neurological, degenerative and scar-adhesion processes in joint area, deforming osteoarthrosis of large joints and osteochondrosis. Method of treatment includes introduction of drug mixture by ultraphonoresis for 5 minutes on a field, daily. To obtain said mixture applied are ointment "Indomethacyne", "Chondroxyde", gel "Essaven" in proportion 1:1:1, in dose, expressed in form of 3-10 cm long stripe of each ointment. These components are mixed with 50 mg of medication Karipasim, dissolved in 5 ml of physiological solution and 5 ml 2% solution of Trental. Treatment course includes 15-20 procedures. |
FIELD: chemistry.
SUBSTANCE: method involves preparation of material for enzymatic hydrolysis. Alkaline hydrolysis is carried out with proteolytic enzyme preparations with neutralisation of the obtained solution to pH=7. A salt is added to the obtained enzymatic hydrolysate to a value of not less than 0.1 mol/l. Successive ultrafiltration is carried out, first on a membrane with maximum retention of 50 kD with separation of high-molecular weight impurities, and then on a membrane with maximum retention of 5 kD with separation of low-molecular weight substances. The chondroitin sulphate solution retained at the membrane is washed on the same membrane with distilled water until complete removal of salts. Final washing with distilled water is carried out on a membrane with maxim retention of 50 kD.
EFFECT: invention enables to obtain a chondroitin sulphate preparation with weight ratio of the basic substance.
7 ex
The invention relates to the fishing industry, in particular to methods for chondroitin sulphate from the tissues of aquatic organisms, such as cartilage tissue of fish, muscular muscular bag molluscs and others, and can be used in food, cosmetic industries, in medicine.
The basic properties of chondroitin sulphate, having crucial for its successful use in various areas - high bioavailability, biological compatibility, low toxicity, ability to selective accumulation in the cartilage (Kofuji K., T. Ito, Y. Murata, S. Kawashima Effect of chondroitin sulfate on the biodegradation and drug release of chitosan gel beads in the subcutaneous air pouches of mice // Biological and Pharmaceutical Bulletin. - 2002. - Vol.25, No. 2. - P. 268-271). Listed properties are determined by the chemical structure of molecules chondroitin sulfate, namely the molecular weight, degree and place . (Michelacci Y. M., Dietrich S.R. Structure of chondroitin sulphate from whale cartilage: distribution of 6 - and 4-sulphated oligosaccharides in the polymer chains // International Journal of Biological Macromolecules. -1986. - Vol.8, No. 2. - P. 108-113. Toida So, Amornrut C., Linhardt R. J. Structure and bioactivity of sulfated polysaccharides // Trends in Glycoscience and Glycotechnology.-2003.-Vol.15, No. 81.-P. 29-46).
To obtain chondroitin sulfate is the most widely used well-known method, providing for the dissolution of chondroitin sulphate in the alkaline environment, enzymatic the hydrolysis of proteins, Department of high-molecular carbohydrate fraction deposition of low molecular weight products of protein hydrolysis, remaining in the solution, rinse the resulting sludge and drying of a finished product (Takai M, Kono N. Salmon-origin chondroitin sulfate: European Patent EP 1270599, IPC A61K 31/737. Appl. 15.12.2000; # EP 20000981747; publ. 02.01.2003).
This common technology is implemented by different authors in different ways: change the sequence number of operations, temperature modes, the nature and concentration of reagents used.
The closest technical solution is a method of obtaining chondroitin sulfate using ultrafiltration (Khare A. Century, Houliston S. A., Black T. J. Isolating chondroitin sulfate: USPTO Application 20070166798. - Appl. 14.02.2007; №11/674695; publ. 07/19/2007).
This way of getting chondroitin sulfate includes collection (preparation of raw material, including connective tissue, hydrolysis feedstock proteoliticakimi enzyme preparations for the solution of the hydrolysate and undissolved substances, handling of liquid hydrolysate reagent, including hydroxide ferrous alkali earth metal, with a pH of more than 10 for the deposition of impurities of protein hydrolysate, Department, at least part of the sludge from the solution of the hydrolysate and processing of liquid hydrolysate using membrane with the formation of leachate (permeate) of low-molecular substances and «detainee» concentrate, which contains the macromolecular fraction of chondroitin sulfate. In the patent is proposed to use the membranes with a molecular weight limit of detention 5 to 15 kDa (better from 8 to 10 kDa).
Product obtained in this manner is a concentrate, which among other substances it contains chondroitin sulfate. Thus, the degree of purification of the target product is not high enough.
The proposed method is also based on division of fractions of the hydrolysate with different molecular weights and uses the high molecular weight of molecules of chondroitin sulphate to separate from the low molecular weight products of hydrolysis of proteins on the ultrafiltration membranes.
Technical the result of this method consists in increasing the degree of purification of the target product by using the ability of molecules chondroitin sulfate change much hydrodynamic radius at the change of ionic force of the solution (electrolyte concentration, e.g. NaCl), allowing for more high in comparison with the prototype of the cleaning of the target product through successive ultrafiltration hydrolysate obtained on the membranes with different threshold of propriety.
As raw materials for production of chondroitin sulfate can be used cartilage tissue containing raw material, received in result of processing of various marine life. When using frozen raw previously conducted its .
Prepared raw material is crushed and loaded into the reaction chamber, which provide alkaline, and then and enzymatic hydrolysis.
Hydrolysed cartilage tissue contains various products breakdown of proteins, salts, high polysaccharides (chondroitin sulfate).
Dissolution substances, including proteins and chondroitin sulfate, carried out at a temperature of 25 to 50 C for 3 h at constant stirring.
After the end of alkaline hydrolysis mixture neutralized to pH 7 and separate sediment filtering or tsentrifugirovaniem.
Holding of alkaline hydrolysis provides early separation impurities and, consequently, increasing the output of the target product, and to improve its purity.
The received solution add an enzyme (FP) or pre-cooked solution OP proteolytic activity, such as enzyme obtained from Kamchatka crab.
Enzymatic hydrolysis of proteins is carried out at optimal for this OP temperature of incubation mixture and duration of treatment (using OP of Kamchatka - temperature of 45 to 55 C and the duration of the hydrolysis of 4 to 8 h), separate the solid residue.
In the resulting solution added salt such as sodium chloride, bringing the salt concentration of 0.1 mol.
Then spend ultrafiltration solution through the membrane from the molecular-mass limit of retaining less than 50 kDa for the Department remaining after hydrolysis of macromolecular proteins and suspended particles.
Concentrated solution containing chondroitin sulfate, wash solution of salts, such as sodium chloride or salt with a concentration of 0.1 mol/L.
For this, the resulting solution added sodium chloride or other salt to maintain its concentration in the solution to the value of not less than 0,1 mol/L. If after the neutralization of the alkali concentration of NaCl above, the additional amount of sodium chloride is added.
When the concentration of NaCl in the solution above 0,1 mol/l molecules chondroitin sulfate strongly that can not distinguish the components of the hydrolysate.
Used salt concentration provides a reduction of hydrodynamic radius of molecules of chondroitin sulfate and the possibility of their passing through the membrane molecular-mass limit of retaining less than 50 kDa, on the membrane linger not hydrolysed proteins such as collagen.
The resulting solution of chondroitin sulphate, low-molecular peptides, amino acids and salts subjected separation by membrane with molecular-mass limit of detention 5 kDa, providing for the detention of molecules of chondroitin sulfate and separation of molecules salts, amino acids and low-molecular peptides.
It is known that lowering the concentration of salt in the solution less than 0.001 mol/l, such as NaCl, molecules chondroitin sulfate are deployed, thus increasing the hydrodynamic radius. Maximum range is observed in distilled water.
Withheld in membrane chondroitin sulfate washed with distilled water resulting concentration of salt is reduced and molecules chondroitin sulfate, hydrodynamic radius of which has substantially grown, can be focused on the membranes with a molecular weight limit of retaining 50 kDa.
On the membrane with a limit of detention 50 kDa chondroitin sulfate finally washed with distilled water from the remaining average molecular weight peptides and implement concentration of its solution by ultrafiltration.
Macromolecular fraction of chondroitin sulfate concentrated on the membrane and low-molecular peptides and amino acids pass through it.
The concentrated solution of chondroitin sulfate is then used to highlight the dry product, or as a solution when making preparations with chondroitin sulfate.
Selection of dry sulfate deposition is adding excess precipitator (e.g., ethyl alcohol) or drying (, spray drying and others).
For example, the resulting solution is precipitated by the addition of alcohol, in a ratio of 1:2, withstand a full sedimentation chondroitin sulfate, separate sludge filtering or tsentrifugirovaniem, wash out with alcohol, acetone, freeze-dried in the dryer, vacuum dryer or other.
Receive drug purified chondroitin sulfate total mass fraction of the basic substance is not less than 90%.
Target product - chondroitin sulfate is a white amorphous powder, odorless, hygroscopic, fraction of total water mass not more than 10%mass fraction of chondroitin sulfate is 90-95%
The definition of a mass fraction of the basic substance is carried out by acid hydrolysis in hydrochloric acid (26% Hcl, 100 C, 1 h) and determine formed glukuronova acid by .
Identification was performed by infrared spectroscopy. As a standard used drug chondroitin 6-sulfate sodium salt of shark cartilage (Catalogue Fluka», cat. no 27043-1G-F).
The results of the comparison showed that samples of chondroitin sulphate obtained by the present method, have almost similar rates to those of the standard sample, which confirms the high degree of purification of the target product.
The use of ultrafiltration is possible when allocating drug chondroitin sulfate after enzymatic hydrolysis of raw materials or when cleaning the besieged ethanol drug, after its dissolution in water.
Examples of the method
Example 1
Getting chondroitin sulfate of cartilage salmon
1) Raw materials - nasal cartilage salmon removed from the goals, cleared from the rests of the surrounding tissues, .
2) 400 g of crushed material is loaded into a flask with 1600 grams of NaOH solution (0.2 mol/dm3 ), heated to boiling water bath for up to 37 C and at this temperature for 3 h at constant stirring held dissolution substances.
3) After the end of the process of mixture neutralized to a pH of 7 using 0.1 N. acetic acid.
4) Separated not residue on the centrifuge at 5000 rpm -1 .
Used enzyme preparation was obtained from Kamchatka crab Paralithodes Camtschaticus by known techniques (Sakharov, I.YU Way to obtain collagenase. /.. .., .., .., .., ... Institute of immunology and Pacific Institute of Bioorganic chemistry: A.S. SU 1343591 A1, 4 61 35/56. - Appl. 13.12.85; №3992368 /28-14. - 2 sec) and had a proteolytic activity on sodium And=280 umol tyrosine*-1 *min-1 . (Substrate: 1%solution of sodium Caseinate. Enzyme: 1 mg/ml solution OP. Conditions of incubation: 37 C, 10 minutes)
6) Held ultrafiltration solution through the membrane 50 kDa. Washed concentrated solution of sodium chloride solution with a concentration of 0.1 mol/l (For the increase of ionic force of the hydrolysate added 9.36 g sodium chloride.)
7) Then the resulting solution of chondroitin sulphate, low-molecular peptides, amino acids and salts subjected separation by membrane 5 kDa, providing detainment of molecules of chondroitin sulfate.
8) Rinsing with distilled water. Solution chondroitin finally got on the membrane 50 kDa from the remaining average molecular weight peptides.
9) Finally, the solution chondroitin sulfate concentrated ultrafiltration.
10) the Resulting solution was besieged by the addition of alcohol, in a ratio of 1:2, survived a solution for 20 hours.
11) Separated sludge by centrifugation at 5000 rpm -1 , washed in 100 ml of alcohol.
12) the Resulting substance dried in the freeze dryer.
Got 6.15 g dry chondroitin sulfate, mass fraction of the basic substance, defined by its reaction with (method ), 91%.
Example 2
Getting chondroitin sulphate from shark cartilage
The same as in example 1, but in quality of raw material used shark cartilage.
Got 6.15 g dry chondroitin sulfate, mass fraction of the basic substance is 93%.
Example 3
Getting chondroitin sulfate from the cartilage of the North slope
The same as in example 1, but in quality of raw material used cartilage of the North slope. Carried out a preliminary degreasing raw materials acetone. In paragraph 5 of enzymatic hydrolysis conducted in two stages for 3 hours with the addition of enzyme preparation of the corresponding concentration.
Got 5,43 g of dry chondroitin sulfate, mass fraction of the basic substance is 92%.
Example 4
Cleaning of chondroitin sulphate of cartilage salmon
Chondroitin sulfate received by way of example 1, except for paragraphs 6-9, again dissolved in 500 ml of distilled water, added 2,93 grams of sodium chloride, the solution is stirred it up within 60 minutes Then held processing solution, as described in example 1 of p.6-9 and next 10-12.
Got to 5.85 g of dry chondroitin sulfate, mass fraction of the basic substance is 94%.
Example 5
Getting chondroitin sulfate of cartilage salmon
The same as in example 1, but as lye operations p.2) instead of NaOH used potassium hydroxide KOH (of 0.2 mol/dm3 ). When ultrafiltration 6) instead of NaCl used 0,1 mol/dm3 solution of potassium chloride KCl.
Got 6,10 g of dry chondroitin sulfate, mass fraction of the basic substance is 92%.
Example 6
Getting chondroitin sulfate of cartilage salmon
The same as in example 1, but instead of NaCl at UF 6) use of 0.05 mol/dm3 solution of calcium chloride.
Got 6.15 g dry chondroitin sulfate, mass fraction of the basic substance is 91%.
Example 7
Getting chondroitin sulfate of cartilage salmon
The same as in example 1, but instead of enzyme preparation from Kamchatka crab used protosubtilin G3X based 4.5 g on 1 kg of raw materials (activity G3X in relation to fisheries squirrel was 1.5 times lower than the activity of AF of Kamchatka crab).
Got 6,35 g of dry chondroitin sulfate, mass fraction of the basic substance is 89%.
The invention allows to receive the drug chondroitin sulfate total mass fraction of the basic substance is not less than 90% (90-95%), and increase the output of the target product.
Method of obtaining of chondroitin sulphate from the tissues of aquatic organisms in providing for the preparation of raw materials to enzymatic hydrolysis, hydrolysis proteoliticakimi enzyme preparations with deposition of protein impurities and the separation of sludge from the solution of the hydrolysate, the selection of the product by means of ultrafiltration, wherein the previously conducted alkaline hydrolysis with the neutralization of the resulting solution to pH 7 and Department of solid residue, enzymatic hydrolysate add salt to the value of not less than 0.1 mol/l, conduct ultrafiltration enzymatic hydrolysate on the membrane with a limit of detention 50 kDa and the Department of high-molecular impurities, resulting solution at a concentration of salt in it not less than 0,1 mol/l subjected ultrafiltration membrane with a limit of detention 5 kDa and a separate low-molecular substances withheld on the membrane chondroitin sulfate is washed on the same membrane distilled water to remove salts, followed by rinsing with distilled water and concentration of a solution of chondroitin sulfate by ultrafiltration membrane with a limit of detention 50 kDa.