1. A process for improving the clarity of chondroitin sulfate is characterized by comprising the following steps:

(1) raw material extraction: mixing 100 parts of chicken bone residues and 350 parts of tap water, heating the mixture by using a pressure tank after mixing, standing the mixture for 15 hours, and performing primary extraction at 115-120 ℃ for 60-80 min; adding the same amount of water for secondary extraction at 130-135 ℃ for 50-60min, and stopping stirring after 1 h;

(2) primary enzymolysis: fishing oil before enzymolysis, adding 2709 protease for enzymolysis, adjusting pH to 8.5, keeping temperature at 54 deg.C, performing enzymolysis for 4h, adjusting pH to 6.2 with citric acid, standing, heating to 90 deg.C, and standing for 10 min;

(3) adsorption: adjusting the pH value to 7.2, adjusting the temperature to 50-60 ℃, adding 10-20 parts of resin, and adsorbing for 5 hours to obtain an eluent;

(4) and (3) oxidation: soaking the collected resin in saline water, collecting all eluates, adding hydrogen peroxide, adjusting pH to 10.5, oxidizing for 4h, and filtering the oxidized liquid with plate-and-frame filter cloth after oxidation;

(5) secondary enzymolysis: adding bromelain and papain to carry out secondary enzymolysis, wherein the mass ratio of the bromelain to the papain is 2:1, the addition amount of the bromelain to the papain is 0.5 percent of the mass of the oxidation liquid, adjusting the temperature to 60-65 ℃, and carrying out enzymolysis on the oxidation liquid for 2 hours;

(6) alkali treatment: firstly, adjusting the pH value to 7-9 with sodium bicarbonate, stirring for 10min, and standing for 1.5 h; adjusting the pH value of the solution to 11-12 by using 30% alkali liquor, stirring for 10min, standing for 1.5h, and filtering by using filter cloth to remove alkali-insoluble impurities;

(7) acid treatment: adjusting the pH value of the filtrate obtained in the step (6) to 2-3 by using 60% lactic acid, stirring for 10min, standing, and removing acid-insoluble impurities by using filter cloth;

(8) alcohol precipitation: carrying out alcohol precipitation by using an alcohol solution with the mass fraction of 74%, and extracting alcohol after 8 hours of alcohol precipitation;

(9) and (3) dehydrating: dehydrating by using a centrifugal machine, and drying in a vacuum drying oven;

(10) and (3) finished product: and cooling to obtain the chondroitin sulfate finished product.

2. The process for improving the clarity of chondroitin sulfate as claimed in claim 1, wherein the resin in the step (3) is Rohm and Haas resin, and the resin is 18 parts.

3. The process for improving the clarity of chondroitin sulfate as claimed in claim 1, wherein the amount of hydrogen peroxide added in the step (4) is 2% of the mass of the eluent.

4. The process for improving the clarity of chondroitin sulfate as claimed in claim 1, wherein the temperature in the vacuum drying oven in the step (9) is 55-60 ℃.

Background

Chondroitin Sulfate (CHS) is a kind of mucopolysaccharide of animal, and is prepared from cartilage tissue containing Chondroitin Sulfate, such as animal larynx, nasal cartilage, trachea or bone tendon, ligament, etc. In cartilage, chondroitin sulfate is bound to proteins (e.g., collagen) and exists in the form of proteoglycans. The chondroitin sulfate is prepared by enzymolysis with alkaline protease 2709 and resin adsorption. The chondroitin sulfate and the protein are combined in a covalent bond mode, and due to the limitation of 2709 protease, some proteins, nucleic acids and the like in the material are remained on the chondroitin. And insoluble impurities with other properties are added, so that the purity of the chondroitin sulfate is not high, the clarity is unqualified, and the product sale is seriously influenced. The method adopts a new process of secondary enzymolysis and acid-base insoluble impurity removal, so that the clarity and purity of the chondroitin sulfate can be improved, and particularly the clarity is obviously improved.

Disclosure of Invention

In order to solve the problems, the invention provides a process for improving the clarity of chondroitin sulfate, which improves the clarity and purity of the chondroitin sulfate, obtains high-quality chondroitin sulfate and facilitates the product to enter high-end markets.

In order to realize the functions, the invention comprises the following steps: (1) raw material extraction: mixing 100 parts of chicken bone residues and 350 parts of tap water, heating the mixture by using a pressure tank after mixing, standing the mixture for 15 hours, and performing primary extraction at 115-120 ℃ for 60-80 min; adding the same amount of water for secondary extraction at 130-135 ℃ for 50-60min, and stopping stirring after 1 h;

(2) primary enzymolysis: fishing oil before enzymolysis, adding 2709 protease for enzymolysis, adjusting pH to 8.5, keeping temperature at 54 deg.C, performing enzymolysis for 4h, adjusting pH to 6.2 with citric acid, standing, heating to 90 deg.C, and standing for 10 min;

(3) adsorption: adjusting the pH value to 7.2, adjusting the temperature to 50-60 ℃, adding 10-20 parts of resin, and adsorbing for 5 hours to obtain an eluent;

(4) and (3) oxidation: soaking the collected resin in saline water, collecting all eluates, adding hydrogen peroxide, adjusting pH to 10.5, oxidizing for 4h, and filtering the oxidized liquid with plate-and-frame filter cloth after oxidation;

(5) secondary enzymolysis: adding bromelain and papain to carry out secondary enzymolysis, wherein the mass ratio of the bromelain to the papain is 2:1, the addition amount of the bromelain to the papain is 0.5 percent of the mass of the oxidation liquid, adjusting the temperature to 60-65 ℃, and carrying out enzymolysis on the oxidation liquid for 2 hours;

(6) alkali treatment: firstly, adjusting the pH value to 7-9 with sodium bicarbonate, stirring for 10min, and standing for 1.5 h; adjusting the pH value of the solution to 11-12 by using 30% alkali liquor, stirring for 10min, standing for 1.5h, and filtering by using filter cloth to remove alkali-insoluble impurities;

(7) acid treatment: adjusting the pH value of the filtrate obtained in the step (6) to 2-3 by using 60% lactic acid, stirring for 10min, standing, and removing acid-insoluble impurities by using filter cloth;

(8) alcohol precipitation: carrying out alcohol precipitation by using an alcohol solution with the mass fraction of 74%, and extracting alcohol after 8 hours of alcohol precipitation;

(9) and (3) dehydrating: dehydrating by using a centrifugal machine, and drying in a vacuum drying oven;

(10) and (3) finished product: and cooling to obtain the chondroitin sulfate finished product.

Further, the resin in the step (3) is a Rohm and Haas resin, and the resin is 18 parts.

Further, the adding amount of hydrogen peroxide in the step (4) is 2% of the mass of the eluent.

Further, the temperature in the vacuum drying oven in the step (9) is 55-60 ℃.

The invention adopts the structure to obtain the following beneficial effects: the process for improving the clarity of the chondroitin sulfate provided by the invention is simple to operate, is different from the traditional preparation process mainly through three processes of secondary enzymolysis, alkali treatment and acid treatment, improves the clarity and purity of the chondroitin sulfate, obtains high-quality chondroitin sulfate, and facilitates the product to enter the high-end market.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1:

the invention relates to a process for improving the clarity of chondroitin sulfate, which comprises the following production processes: (1) raw material extraction: mixing 100 parts of chicken bone residues and 350 parts of tap water, heating the mixture by using a pressure tank after mixing, standing the mixture for 15 hours, and performing primary extraction at 115-120 ℃ for 60-80 min; adding the same amount of water for secondary extraction at 130-135 ℃ for 50-60min, and stopping stirring after 1 h;

(2) primary enzymolysis: fishing oil before enzymolysis, adding 2709 protease for enzymolysis, adjusting pH to 8.5, keeping temperature at 54 deg.C, performing enzymolysis for 4h, adjusting pH to 6.2 with citric acid, standing, heating to 90 deg.C, and standing for 10 min;

(3) adsorption: adjusting the pH value to 7.2, adjusting the temperature to 50-60 ℃, adding 10-20 parts of resin, and adsorbing for 5 hours to obtain an eluent;

(4) and (3) oxidation: soaking the collected resin in saline water, collecting all eluates, adding hydrogen peroxide, adjusting pH to 10.5, oxidizing for 4h, and filtering the oxidized liquid with plate-and-frame filter cloth after oxidation;

(5) secondary enzymolysis: adding bromelain and papain to carry out secondary enzymolysis, wherein the mass ratio of the bromelain to the papain is 2:1, the addition amount of the bromelain to the papain is 0.5 percent of the mass of the oxidation liquid, adjusting the temperature to 60 ℃, and carrying out enzymolysis on the oxidation liquid for 2 hours;

(6) alkali treatment: firstly, adjusting the pH value to 8 by using sodium bicarbonate, stirring for 10min, and standing for 1.5 h; adjusting the pH value of the solution to 11 by using 30% alkali liquor, stirring for 10min, standing for 1.5h, and filtering by using filter cloth to remove alkali insoluble impurities;

(7) acid treatment: adjusting the pH value of the filtrate obtained in the step (6) to 2 by using 60% lactic acid, stirring for 10min, standing, and removing acid-insoluble impurities by using filter cloth;

(8) alcohol precipitation: carrying out alcohol precipitation by using an alcohol solution with the mass fraction of 74%, and extracting alcohol after 8 hours of alcohol precipitation;

(9) and (3) dehydrating: dehydrating by using a centrifugal machine, and drying at 58 ℃ in a vacuum drying oven;

(10) and (3) finished product: and cooling to obtain the chondroitin sulfate finished product.

Example 2:

the invention relates to a process for improving the clarity of chondroitin sulfate, which comprises the following production processes: (1) raw material extraction: mixing 20-100 parts of chicken bone residues and 100-500 parts of tap water, heating by using a pressure tank after mixing, standing for 15 hours, and performing primary extraction at 115-120 ℃ for 60-80 min; adding the same amount of water for secondary extraction at 130-135 ℃ for 50-60min, and stopping stirring after 1 h;

(2) primary enzymolysis: fishing oil before enzymolysis, adding 2709 protease for enzymolysis, adjusting pH to 8.5, keeping temperature at 54 deg.C, performing enzymolysis for 4h, adjusting pH to 6.2 with citric acid, standing, heating to 90 deg.C, and standing for 10 min;

(3) adsorption: adjusting the pH value to 7.2, adjusting the temperature to 50-60 ℃, adding 10-20 parts of resin, and adsorbing for 5 hours to obtain an eluent;

(4) and (3) oxidation: soaking the collected resin in saline water, collecting all eluates, adding hydrogen peroxide, adjusting pH to 10.5, oxidizing for 4h, and filtering the oxidized liquid with plate-and-frame filter cloth after oxidation;

(5) secondary enzymolysis: adding bromelain and papain to carry out secondary enzymolysis, wherein the mass ratio of the bromelain to the papain is 2:1, the addition amount of the bromelain to the papain is 0.5 percent of the mass of the oxidation liquid, adjusting the temperature to 65 ℃, and carrying out enzymolysis on the oxidation liquid for 2 hours;

(6) alkali treatment: firstly, adjusting the pH value to 9 by using sodium bicarbonate, stirring for 10min, and standing for 1.5 h; adjusting the pH value of the solution to 11.5 by using 30% alkali liquor, stirring for 10min, standing for 1.5h, and filtering by using filter cloth to remove alkali insoluble impurities;

(7) acid treatment: adjusting the pH value of the filtrate obtained in the step (6) to 3 by using 60% lactic acid, stirring for 10min, standing, and removing acid-insoluble impurities by using filter cloth;

(8) alcohol precipitation: carrying out alcohol precipitation by using an alcohol solution with the mass fraction of 74%, and extracting alcohol after 8 hours of alcohol precipitation;

(9) and (3) dehydrating: dehydrating by using a centrifugal machine, and drying in a vacuum drying oven at 55 ℃;

(10) and (3) finished product: and cooling to obtain the chondroitin sulfate finished product. .

Example 3:

the invention relates to a process for improving the clarity of chondroitin sulfate, which comprises the following production processes: (1) raw material extraction: mixing 100 parts of chicken bone residues and 350 parts of tap water, heating the mixture by using a pressure tank after mixing, standing the mixture for 15 hours, and performing primary extraction at 115-120 ℃ for 60-80 min; adding the same amount of water for secondary extraction at 130-135 ℃ for 50-60min, and stopping stirring after 1 h;

(2) primary enzymolysis: fishing oil before enzymolysis, adding 2709 protease for enzymolysis, adjusting pH to 8.5, keeping temperature at 54 deg.C, performing enzymolysis for 4h, adjusting pH to 6.2 with citric acid, standing, heating to 90 deg.C, and standing for 10 min;

(3) adsorption: adjusting the pH value to 7.2, adjusting the temperature to 50-60 ℃, adding 10-20 parts of resin, and adsorbing for 5 hours to obtain an eluent;

(4) and (3) oxidation: soaking the collected resin in saline water, collecting all eluates, adding hydrogen peroxide, adjusting pH to 10.5, oxidizing for 4h, and filtering the oxidized liquid with plate-and-frame filter cloth after oxidation;

(5) secondary enzymolysis: adding bromelain and papain to carry out secondary enzymolysis, wherein the mass ratio of the bromelain to the papain is 2:1, the addition amount of the bromelain to the papain is 0.5 percent of the mass of the oxidation liquid, adjusting the temperature to 63 ℃, and carrying out enzymolysis on the oxidation liquid for 2 hours;

(6) alkali treatment: firstly, adjusting the pH value to 8 by using sodium bicarbonate, stirring for 10min, and standing for 1.5 h; adjusting the pH value of the solution to 12 by using 30% alkali liquor, stirring for 10min, standing for 1.5h, and filtering by using filter cloth to remove alkali insoluble impurities;

(7) acid treatment: adjusting the pH value of the filtrate obtained in the step (6) to 2.5 by using 60% lactic acid, stirring for 10min, standing, and removing acid-insoluble impurities by using filter cloth;

(8) alcohol precipitation: carrying out alcohol precipitation by using an alcohol solution with the mass fraction of 74%, and extracting alcohol after 8 hours of alcohol precipitation;

(9) and (3) dehydrating: dehydrating by using a centrifugal machine, and drying in a vacuum drying oven at 60 ℃;

(10) and (3) finished product: and cooling to obtain the chondroitin sulfate finished product.

The invention and its embodiments have been described above, without this being limitative. In summary, those skilled in the art should appreciate that they can readily use the disclosed conception and specific embodiments as a basis for designing or modifying other structures for carrying out the same purposes of the present invention without departing from the spirit and scope of the invention as defined by the appended claims.