CN101358220A

CN101358220A - Method for extracting calcium chondroitin sulfate in shark cartilage

Info

Publication number: CN101358220A
Application number: CNA2008101210533A
Authority: CN
Inventors: 王清荣; 傅应华; 罗程; 杨明富
Original assignee: Individual
Current assignee: Individual
Priority date: 2008-09-25
Filing date: 2008-09-25
Publication date: 2009-02-04

Abstract

A method for extracting shark calcium chondroitin sulfate from a shark cartilage comprises the processes in sequence such as curing, alkalization, enzymatic hydrolysis, deproteinization, crystallization, resin pretreatment, resin exchange, ultrafiltration dehydration, calcification of filtrate and crystallization dehydration dying. Because secondary crystallization is adopted, the covalent binding of protein and shark chondroitin sulfate is broken by hydrogen peroxide to enable the chondroitin sulfate in the protein to be dissociated, impurities such as the protein and the like can be effectively removed to improve the purity and the content of shark sodium chondroitin sulfate; cation resin is adopted to complete the sodium removal process by column chromatography, the exchanged shark chondroitin sulfate is high in concentration and the shark chondroitin sulfate which is produced during ion exchange can be accurately controlled, and the sodium content in the product is low; with dehydration of ultrafiltration, not only the concentration can be realized, but also the small molecular impurities can be removed to improve the purity of the shark chondroitin sulfate, at the same time, to improve the content of calcium. The method also can save the using amount of ethanol and reduce the production cost.

Description

Extract the method for calcium chondroitin sulfate in the shark suft bone

Technical field

The present invention relates to the preparation method of shark chondroitine calcium, particularly a kind of method of from shark suft bone, extracting shark chondroitine calcium

Background technology

Chondroitin sulfate gets important component part as reticular tissue, has multiple pharmacological effect and biological function, often is used as dietary supplements and uses, and is mainly used in the assisting therapy of osteoarthritis.The traditional method that chondroitin sulfate extracts mainly contains alkali and puies forward enzymolysis process, alkaline extraction, ultrasonic wave auxiliary law and acetic acid extraction method, the chondroitin sulfate of making by traditional method mainly exists with the form of sodium salt, because chondroitin sulfate is stronger than sodium to the affinity of calcium, after the use, the calcium that often causes blood vessel and capillary vessel position stockpiles and causes petechiae and hemotoncus, common alkaline process production technique to have also that purity is low, the ethanol usage quantity causes the high defective of production cost greatly.

Summary of the invention

The objective of the invention is to propose a kind ofly can overcome above-mentioned defective, chondroitin sulfate is changed into the method for extracting the high purity calcium chondroitin sulfate in the shark suft bone of calcium salt by sodium salt.

Technical scheme of the present invention is: the method for extracting shark chondroitine calcium in a kind of shark suft bone, include slaking, alkalization, enzymolysis, Deproteinization, crystallization, resin pre-treatment, resins exchange, ultrafiltration dehydration, filtrate calcification, dry each the operation process composition of exsiccation in regular turn, above-mentioned:

Slaking: get shark suft bone, under neutrallty condition, 80-90 degree centigrade poach 1-3 hour, wash down and remove grease and add water boil again, be incubated 2-5 hour;

Alkalization: the 2%-10% that presses the shark suft bone charging capacity adds sodium hydroxide, and the 3%-10% that presses the shark suft bone charging capacity adds sodium-chlor, is heated to 30-40 degree centigrade, stirs alkalization 1-5 hour, treats to stop alkalization after the chondrolysis;

Enzymolysis: transferring pH value with hydrochloric acid is 8-10, and temperature 40-60 degree centigrade, the 0.1%-0.4% that presses the cartilage charging capacity adds pancreatin, and enzymolysis process is constantly added sodium hydroxide;

Deproteinization: adjusting pH value with hydrochloric acid is 7-9, is warming up to 80-100 degree centigrade, is incubated 1-3 hour, filters, and obtains settled solution;

It is characterized in that: secondary crystal is adopted in above-mentioned crystallization, in the clear liquor of Deproteinization, add 95% ethanol earlier, make alcohol concn reach 70%, crystallization is filtered and is obtained the shark chondroitine raw product, be dissolved in water again, add 1-5% sodium-chlor again, transfer pH value to 7-9 with sodium hydroxide, add hydrogen peroxide, elevated temperature is to 40-70 degree centigrade, and oxidization time was controlled at 3-8 hour, transfers pH value to 2-5 again, filter, get clear filtrate, transfer pH value to 6-7, add 95% ethanol with sodium hydroxide, make alcohol concn reach 70%, crystallization is filtered and is obtained the shark chondroitine highly finished product; Above-mentioned resin pre-treatment is to use 732 strongly acidic styrene type cation exchange resins, uses water rinse, soaks, and with 1 mol sodium hydroxide alkali cleaning, with 1 mol chlorohydric acid pickling, water washes to the about 6-7 of pH value again, by wet method dress post; Above-mentioned resins exchange is to get the shark chondroitine highly finished product, be dissolved in water by weight, make the solution of 5%-15%, upper prop is controlled take-off rate well, and with the pH value of the online detection effluent liquid of acidometer, when the pH value of effluent liquid becomes 4 by neutrality, begin to collect effluent liquid, when pH value becomes 4 by strong acid, stop to collect; Above-mentioned ultrafiltration dehydration is to get above-mentioned ion exchanged soln, and the ultrafiltration dehydration is sloughed molecular weight simultaneously less than impurity such as 1000 polypeptide, amino acid, improves shark chondroitine purity; Above-mentioned filtrate calcification is the concentrated solution of getting after ultrafiltration is dewatered, and regulates pH value to 3-8 with calcium hydroxide water saturation liquid, adds calcium chloride again, and stirring and dissolving is filtered.

Extract the method for calcium chondroitin sulfate in the shark suft bone of the present invention owing to adopt secondary crystal, covalent attachment with hydrogen peroxide destroying protein and shark chondroitine, make the chondroitin sulfate in the protein dissociate out, can effectively remove impurity such as protein, improve the purity and the content of shark chondroitine sodium, adopt Zeo-karb, finish the sodium process with column chromatography, the shark chondroitine concentration height of exchange, and can accurately control the shark chondroitine that generates in the ion exchange process, sodium content is low in the product, the hyperfiltration process dehydration both can have been played concentrated, can slough small molecular weight impurity simultaneously again, improve the purity of shark chondroitine, can improve calcium content simultaneously.Also can save the alcoholic acid usage quantity, reduce production costs.

Embodiment

The present invention is further illustrated below in conjunction with embodiment:

Get shark suft bone 200 grams, add water, be heated to 90 degrees centigrade, boiled 2 hours, after taking out the flushing of cartilage water, added water boil again 4 hours, add 10 gram sodium hydroxide, 10 gram sodium-chlor, 40 degree stir alkalization 5 hours down, transfer pH value to 8.5 with hydrochloric acid, 55 degrees centigrade of heating and controlled temperature, add pancreatin 0.6 gram, add at twice, add for the first time 0.4 gram, add 0.2 gram after three hours, constantly stir, enzyme digestion reaction 6 hours, adjusting pH value with hydrochloric acid is 7.5, be warming up to 80 degrees centigrade, be incubated 2 hours, filter clear filtrate, add 95% ethanol, make alcohol concn reach 70%, stirred once every 30 minutes, crystallization is filtered and is obtained the shark chondroitine raw product, weighing 80 grams, add 480 milliliters of dissolvings of water again, add 4.8 gram sodium-chlor again, transfer pH value to 8.5, elevated temperature to 50 degree centigrade with sodium hydroxide, add 4.8 milliliters of hydrogen peroxide, oxidizing reaction 5 hours is transferred pH value to 2.5 with hydrochloric acid again, filters, get clear filtrate, transfer pH value to 6.5 with sodium hydroxide, add 95% ethanol, make alcohol concn reach 70%, stirred once every 30 minutes, crystallization is filtered and is obtained the shark chondroitine highly finished product, with 500 milliliters of hot water dissolving's throw outs, uses 732 Zeo-karbs, with the online detection pH value of acidometer, when the pH value of effluent liquid becomes 4 by neutrality, begin to collect effluent liquid, when pH value becomes 4 by strong acid, stop to collect, hold back greater than 1000 daltonian chondroitin sulfate acid through ultrafiltration, dehydration is 300 milliliters after concentrating again, and regulates pH value to 6 with calcium hydroxide water saturation liquid, add calcium chloride 140 grams again, stirring and dissolving is filtered, and precipitation is dewatered with dehydrated alcohol, 65 degrees centigrade of vacuum-dryings, get shark chondroitine calcium 50 grams, purity reaches 98% after testing, and all other indexs meet technical requirements.

Claims

1, extracts the method for shark chondroitine calcium in a kind of shark suft bone, include slaking, alkalization, enzymolysis, Deproteinization, crystallization, resin pre-treatment, resins exchange, ultrafiltration dehydration, filtrate calcification, dry each the operation process composition of exsiccation in regular turn, above-mentioned:

It is characterized in that: secondary crystal is adopted in above-mentioned crystallization, in the clear liquor of Deproteinization, add 95% ethanol earlier, make alcohol concn reach 70%, crystallization is filtered and is obtained the shark chondroitine raw product, be dissolved in water again, add 1-5% sodium-chlor again, transfer pH value to 7-9 with sodium hydroxide, add hydrogen peroxide, elevated temperature is to 40-70 degree centigrade, and oxidization time was controlled at 3-8 hour, transfers pH value to 2-5 again, filter, get clear filtrate, transfer pH value to 6-7, add 95% ethanol with sodium hydroxide, make alcohol concn reach 70%, crystallization is filtered and is obtained the shark chondroitine highly finished product.

2, extract the method for shark chondroitine calcium in a kind of shark suft bone according to claim 1, it is characterized in that above-mentioned resin pre-treatment is to use 732 strongly acidic styrene type cation exchange resins, use water rinse, soak, with 1 mol sodium hydroxide alkali cleaning, with 1 mol chlorohydric acid pickling, water washes to the about 6-7 of pH value again, by wet method dress post.

3, extract the method for shark chondroitine calcium in a kind of shark suft bone according to claim 1, it is characterized in that above-mentioned resins exchange is to get the shark chondroitine highly finished product, be dissolved in water by weight, make the solution of 5%-15%, upper prop, control take-off rate well, and, when the pH value of effluent liquid becomes 4 by neutrality, begin to collect effluent liquid with the pH value of the online detection effluent liquid of acidometer, when pH value becomes 4 by strong acid, stop to collect.

4, extract the method for shark chondroitine calcium in a kind of shark suft bone according to claim 1, it is characterized in that above-mentioned ultrafiltration dehydration is to get above-mentioned ion exchanged soln, the ultrafiltration dehydration is sloughed molecular weight simultaneously less than impurity such as 1000 polypeptide, amino acid.

5, extract the method for shark chondroitine calcium in a kind of shark suft bone according to claim 1, it is characterized in that above-mentioned filtrate calcification is the concentrated solution of getting after ultrafiltration is dewatered, regulate pH value to 3-8, add calcium chloride again with calcium hydroxide water saturation liquid, stirring and dissolving is filtered.