Method for preparing high-purity chondroitin sulfate

Abstract

The invention discloses a method for preparing chondroitin sulfate, which comprises the following steps: boiling animal cartilage, performing enzymolysis and filtering to obtain enzymolysis liquid; subjecting the enzymolysis liquid to ultrafiltration concentration to obtain primary concentrate; passing the primary concentrate through cation exchange resin, and collecting effluent; washing the cation exchange resin with water, and collecting and mixing effluent; subjecting the effluent to ultrafiltration concentration to obtain secondary concentrate; and precipitating the secondary concentrate in ethanol, dewatering and drying to obtain chondroitin sulfate. The method disclosed by the invention has the advantages of high selectivity, high product purity, high yield, low energy consumption, low ethanol consumption, mild process conditions, simple operation and the like.

Description

(1) Technical field

[0001] The invention relates to a method for preparing high-purity chondroitin sulfate, in particular to a method for directly separating and purifying chondroitin sulfate from cartilage enzymatic hydrolysis solution. (2) Background technology

[0002] Chondroitin Sulfate (CS) is a kind of acidic mucopolysaccharide widely distributed in animal cartilage tissue. Its molecular structure is a polysaccharide composed of repeating disaccharide units composed of D-glucuronic acid and N-acetylgalactosamine. Clinically, it is mainly used to prevent and treat neuralgia, neuropathic migraine, arthritis, shoulder joint pain, post-abdominal surgery pain, coronary heart disease, angina pectoris and myocardial infarction, etc., for hearing impairment caused by streptomycin and hearing impairment caused by various noises. The prevention and treatment of difficulty, tinnitus and other diseases also have good curative effect.

[0003] my country is a large agricultural ...

Examples

Embodiment 1

[0029] 1) Take 20Kg pig nose bone (water content 12%), add 400L water, 4KgNaCl, heat up to 100°C, keep warm for 2h; naturally cool to 45°C-48°C, adjust the pH to 9.0 with a 40% NaOH solution, Add 300g of trypsin for enzymolysis for 6h, after the end of enzymolysis, heat up to 70°C, stir for 30min, filter to obtain 400L of enzymolysis solution;

[0030] 2) Naturally cool the enzymolysis solution to 40°C, adjust the pH to 7.0-8.0 with 6mol / L hydrochloric acid solution, and conduct ultrafiltration concentration with a hollow fiber ultrafilter under a working pressure of 0.15MPa, with a molecular weight cut-off of 3000, to obtain 68L concentrate;

[0031] 3) Pass 68L of concentrated solution through DK110 type cation exchange resin with a volume of 68L at a flow rate of 1.0BV / h, and collect the effluent; then wash the cation exchange resin with 272L of water at a flow rate of 2.0BV / h;

[0032] 4) Combine the effluents, adjust the pH to 7.0-8.0 with a NaOH solution with a mass con...

Embodiment 2

[0035] 1) with embodiment 1

[0036]2) Naturally cool the enzymolysis solution to 40°C, adjust the pH to 7.0-8.0 with 6mol / L hydrochloric acid solution, and conduct ultrafiltration concentration with a hollow fiber ultrafilter under a working pressure of 0.15MPa, with a molecular weight cut-off of 5000, to obtain 33L concentrate;

[0037] 3) Pass 33L of concentrated solution through HD-8 type cation exchange resin with a volume of 66L at a flow rate of 2.0BV / h, and collect the effluent; then wash the cation exchange resin with 132L of water at a flow rate of 3.0BV / h;

[0038] 4) Combine the effluents, adjust the pH to 7.0-8.0 with a NaOH solution with a mass concentration of 40%, and carry out ultrafiltration and concentration with a roll-type ultrafilter under a working pressure of 0.5 MPa. The molecular weight cut-off is 2000, and 22 L of secondary concentration is obtained. liquid;

[0039] 5) Add 66L of 95% ethanol to 22L of the secondary concentrated solution, alcohol p...

Embodiment 3

[0041] 1) with embodiment 1

[0042] 2) Naturally cool the enzymolysis solution to 40°C, adjust the pH to 7.0-8.0 with 6mol / L hydrochloric acid solution, and conduct ultrafiltration concentration with a hollow fiber ultrafilter under a working pressure of 0.15MPa, with a molecular weight cut-off of 6000, to obtain 41L concentrate;

[0043] 3) Pass 41L of the concentrated solution through a C150s type cation exchange resin with a volume of 123L at a flow rate of 0.5BV / h, and collect the effluent; then wash the cation exchange resin with 492L of water at a flow rate of 0.1BV / h;

[0044] 4) Combine the effluents, adjust the pH to 7.0-8.0 with a NaOH solution with a mass concentration of 40%, and carry out ultrafiltration and concentration with a roll-type ultrafilter at a working pressure of 0.5 MPa. The molecular weight cut-off is 3000, and 22 L of secondary concentration is obtained. liquid;

[0045] 5) Add 110L of 95% ethanol to 22L of the secondary concentrated solution, al...

(1) Technical field

[0001] The invention relates to a method for preparing high-purity chondroitin sulfate, in particular to a method for directly separating and purifying chondroitin sulfate from cartilage enzymatic hydrolysis solution. (2) Background technology

[0002] Chondroitin Sulfate (CS) is a kind of acidic mucopolysaccharide widely distributed in animal cartilage tissue. Its molecular structure is a polysaccharide composed of repeating disaccharide units composed of D-glucuronic acid and N-acetylgalactosamine. Clinically, it is mainly used to prevent and treat neuralgia, neuropathic migraine, arthritis, shoulder joint pain, post-abdominal surgery pain, coronary heart disease, angina pectoris and myocardial infarction, etc., for hearing impairment caused by streptomycin and hearing impairment caused by various noises. The prevention and treatment of difficulty, tinnitus and other diseases also have good curative effect.

[0003] my country is a large agricultural ...

Examples

Embodiment 1

[0029] 1) Take 20Kg pig nose bone (water content 12%), add 400L water, 4KgNaCl, heat up to 100°C, keep warm for 2h; naturally cool to 45°C-48°C, adjust the pH to 9.0 with a 40% NaOH solution, Add 300g of trypsin for enzymolysis for 6h, after the end of enzymolysis, heat up to 70°C, stir for 30min, filter to obtain 400L of enzymolysis solution;

[0030] 2) Naturally cool the enzymolysis solution to 40°C, adjust the pH to 7.0-8.0 with 6mol / L hydrochloric acid solution, and conduct ultrafiltration concentration with a hollow fiber ultrafilter under a working pressure of 0.15MPa, with a molecular weight cut-off of 3000, to obtain 68L concentrate;

[0031] 3) Pass 68L of concentrated solution through DK110 type cation exchange resin with a volume of 68L at a flow rate of 1.0BV / h, and collect the effluent; then wash the cation exchange resin with 272L of water at a flow rate of 2.0BV / h;

[0032] 4) Combine the effluents, adjust the pH to 7.0-8.0 with a NaOH solution with a mass con...

Embodiment 2

[0035] 1) with embodiment 1

[0036]2) Naturally cool the enzymolysis solution to 40°C, adjust the pH to 7.0-8.0 with 6mol / L hydrochloric acid solution, and conduct ultrafiltration concentration with a hollow fiber ultrafilter under a working pressure of 0.15MPa, with a molecular weight cut-off of 5000, to obtain 33L concentrate;

[0037] 3) Pass 33L of concentrated solution through HD-8 type cation exchange resin with a volume of 66L at a flow rate of 2.0BV / h, and collect the effluent; then wash the cation exchange resin with 132L of water at a flow rate of 3.0BV / h;

[0038] 4) Combine the effluents, adjust the pH to 7.0-8.0 with a NaOH solution with a mass concentration of 40%, and carry out ultrafiltration and concentration with a roll-type ultrafilter under a working pressure of 0.5 MPa. The molecular weight cut-off is 2000, and 22 L of secondary concentration is obtained. liquid;

[0039] 5) Add 66L of 95% ethanol to 22L of the secondary concentrated solution, alcohol p...

Embodiment 3

[0041] 1) with embodiment 1

[0042] 2) Naturally cool the enzymolysis solution to 40°C, adjust the pH to 7.0-8.0 with 6mol / L hydrochloric acid solution, and conduct ultrafiltration concentration with a hollow fiber ultrafilter under a working pressure of 0.15MPa, with a molecular weight cut-off of 6000, to obtain 41L concentrate;

[0043] 3) Pass 41L of the concentrated solution through a C150s type cation exchange resin with a volume of 123L at a flow rate of 0.5BV / h, and collect the effluent; then wash the cation exchange resin with 492L of water at a flow rate of 0.1BV / h;

[0044] 4) Combine the effluents, adjust the pH to 7.0-8.0 with a NaOH solution with a mass concentration of 40%, and carry out ultrafiltration and concentration with a roll-type ultrafilter at a working pressure of 0.5 MPa. The molecular weight cut-off is 3000, and 22 L of secondary concentration is obtained. liquid;

[0045] 5) Add 110L of 95% ethanol to 22L of the secondary concentrated solution, al...