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Low molecular weight chondroitin sulfate preparation method

A chondroitin sulfate, low molecular weight technology, applied in fermentation and other directions, can solve the problems of unsuitability for large-scale production, wide product molecular weight distribution, high equipment requirements, maintain integrity and biological activity, reduce the difficulty of complex enzymatic hydrolysis, and improve The effect of product quality

Inactive Publication Date: 2015-04-08
厦门蓝湾科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The process requires repeated operations such as purification, precipitation, and drying, which are complicated and costly
In addition, domestic chondroitin sulfate degradation technology research is less, common degradation methods include enzymatic method, acid method, hydrogen peroxide method, etc., but all have their imperfections
The Chinese patent applications whose publication numbers are CN101659973A and CN103602711A all involve the preparation method of low-molecular chondroitin sulfate. Although the enzymatic method has the advantages of less environmental pollution, mild reaction conditions, and high product purity, the enzyme that degrades CS has high specificity and is expensive. Rare sources, not suitable for mass production
The Chinese patent application whose publication number is CN1940082A discloses a method for preparing low-molecular-weight chondroitin sulfate by hydrochloric acid degradation. The production cycle is long and the product purity is not high; Preparation method, however, the degradation mechanism of hydrogen peroxide method is not yet fully clear, the 2011 national food additive use standard (GB2760-2011) has canceled the use of hydrogen peroxide as an auxiliary agent in food processing, and its safety needs further research
The Chinese patent application with publication number CN103554304A relates to a method for preparing low-molecular-weight sturgeon chondroitin sulfate from sturgeon cartilage. Although the degradation of chondroitin sulfate by radiation combined with ultrafine pulverization is simple and easy, and pollution-free, it first obtains large-molecular-weight sulfuric acid Chondroitin, re-degradation to obtain low molecular weight chondroitin sulfate products has complicated process steps, long production cycle, high equipment requirements, and wide molecular weight distribution of the product
So far, the preparation process of low-molecular-weight chondroitin sulfate that is safe, reliable, has industrialization potential, and has low production cost has not yet appeared.

Method used

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  • Low molecular weight chondroitin sulfate preparation method
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  • Low molecular weight chondroitin sulfate preparation method

Examples

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Embodiment 1

[0040] Take fresh sturgeon spine tissue and place it in a boiling water bath for 1 hour, remove muscle, fat and other attachments on the surface of the spine, soak in petroleum ether, wash with alcohol, dry at 70°C, pulverize with a grinder, and weigh for later use. Weigh 30g of cartilage granules, add 4% NaOH solution according to the ratio of material to liquid 1:5, and stir at 45°C for 30min. After adjusting the pH to 7.0 with hydrochloric acid, add malic acid until the pH is 3.0, add pepsin with 0.8% cartilage mass, and stir at 37°C for 4 hours, during which the pH is adjusted to be stable at 3.0. Adjust the pH to 5.0, add papain and cellulase with a mass of 3% cartilage at a ratio of 1:1, stir at 55°C for 4 hours, and adjust the pH to stabilize at 5.0 during this period. Add 0.5% activated carbon in the volume of the solution, stir at 80°C for 30 minutes, cool and filter to obtain the enzymolysis solution. Adjust the pH to 6.0, adjust the NaCl concentration to 0.25mol / L,...

Embodiment 2

[0045] Take fresh sturgeon spine tissue and place it in a boiling water bath for 1 hour, remove muscle, fat and other attachments on the surface of the spine, soak in petroleum ether, wash with alcohol, dry at 70°C, pulverize with a grinder, and weigh for later use. Weigh 30g of cartilage granules, add 4% NaOH solution according to the ratio of material to liquid 1:5, and stir at 45°C for 30min. After adjusting the pH to 7.0 with hydrochloric acid, add malic acid until the pH is 3.0, add pepsin with 1% cartilage mass, and stir at a constant temperature of 37°C for 4 hours, during which the pH is adjusted to be stable at 3.0. Adjust the pH to 5.0, add papain and cellulase with 4% cartilage mass in a ratio of 1.2:1, stir at 55°C for 4 hours, and adjust the pH to stabilize at 5.0 during this period. Add 0.5% activated carbon in the volume of the solution, stir at 80°C for 30 minutes, cool and filter to obtain the enzymolysis solution. Adjust the pH to 6.0, adjust the NaCl concen...

Embodiment 3

[0047] Take fresh sturgeon spine tissue and place it in a boiling water bath for 1 hour, remove muscle, fat and other attachments on the surface of the spine, soak in petroleum ether, wash with alcohol, dry at 70°C, pulverize with a grinder, and weigh for later use. Weigh 30g of cartilage particles, add 6% NaOH solution according to the ratio of material to liquid 1:5, and stir at 50°C for 45min. After adjusting the pH to 7.0 with hydrochloric acid, add malic acid until the pH is 3.0, add pepsin with 1% cartilage mass, and stir at a constant temperature of 37°C for 4 hours, during which the pH is adjusted to be stable at 3.0. Adjust the pH to 5.5, add papain and cellulase with a mass of 4% cartilage at a ratio of 1.4:1, stir at 60°C for 4 hours, and adjust the pH to stabilize at 5.5. Add 0.5% activated carbon by volume of the solution, stir at 90°C for 30 minutes, cool and filter to obtain the enzymatic hydrolysis solution. Adjust the pH to 6.0, adjust the NaCl concentration ...

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Abstract

A low molecular weight chondroitin sulfate preparation method relates to chondroitin sulfate. The method includes the steps of stewing sturgeon to remove flesh and fat on the surface of bone, soaking with petroleum ether, washing with ethanol, drying and pulverizing to obtain cartilage particles; adding a NaOH solution to the cartilage particles, stirring for the first time and adjusting the pH to neutral with acid to obtain a cartilage mother solution; adding malic acid to the cartilage mother solution, adjusting the pH to 3.0 to 3.2, adding pepsin, stirring for the second time, adjusting the pH to 5.0 to 5.5, adding papain and cellulase, stirring for the third time, adding activated carbon, stirring for the fourth time and filtering to obtain clarified enzymatic hydrolysate; subjecting the enzymatic hydrolysate to resin adsorption, gradient elution in a NaCl solution, phloroglucinol spectrophotometry detection and combined collection to obtain an eluent with color reaction; precipitating the eluate with ethanol, collecting precipitate through centrifugation and drying to obtain low molecular weight chondroitin sulfate crystals.

Description

technical field [0001] The invention relates to chondroitin sulfate, in particular to a preparation method of low molecular weight chondroitin sulfate. Background technique [0002] Chondroitin sulfate (CS for short) is a kind of natural acidic mucopolysaccharide from animal cartilage tissue, its sugar chain is composed of alternating disaccharide units (N-glucuronic acid and N-acetylgalactosamine) and connected on the core protein. Studies have shown that chondroitin sulfate plays an important role in the treatment of joint injuries, corneal injuries, neuralgia, anti-inflammatory and antibacterial, and has various biological activities such as promoting coronary circulation, lowering blood lipids, anticoagulation, antiviral and antitumor, etc. It has been widely used as a raw material in the pharmaceutical, health food and cosmetic industries. [0003] my country is the most important sturgeon producing country in the world, and the aquaculture output accounts for more th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/26
Inventor 石国宗潘启胜林颖珺秦飞陈静
Owner 厦门蓝湾科技有限公司
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