Processing technology of sodium chondroitin sulfate

Abstract

The invention relates to the field of food deep processing, and particularly discloses a processing technology of sodium chondroitin sulfate, which comprises the following steps: S1, crushing raw materials, boiling, carrying out alkaline hydrolysis, and carrying out enzymolysis to obtain an enzymatic hydrolysate; s2, the enzymatic hydrolysate is subjected to filtration, ultrafiltration, oxidation, secondary filtration, ultrafiltration membrane filtration, alcohol precipitation, dehydration, centrifugation, drying, crushing and sieving, and a finished product is obtained; the method has the advantages of high extraction rate, high extraction rate and high purity.

Description

Technical Field

[0001] This application relates to the field of food deep processing, and more specifically, it relates to a processing technology for sodium chondroitin sulfate. Background Technology

[0002] Chondroitin sulfate sodium is a sulfated chain-like mucopolysaccharide sodium salt extracted from cartilage tissues such as the laryngeal bone, nasal bone, and trachea of ​​pigs. It has the effects of protecting articular cartilage, promoting cartilage repair, anti-inflammatory and analgesic, improving blood lipid metabolism, as well as anticoagulation, accelerating wound healing, and anti-tumor effects.

[0003] In existing technologies, sodium chondroitin sulfate can be extracted using alkaline methods, enzymatic methods, and extraction methods. Traditional alkaline soaking requires 5-7 days, and the alkaline solution must be replaced every 24 hours to prevent the accumulation of byproducts. Therefore, although the alkaline method has a high extraction rate, the production cycle is long and it is easy to cause excessive environmental pollution. Although the enzymatic method has a high yield and purity, it is easy to introduce other residues. The acetic acid extraction method has a low extraction rate.

[0004] Therefore, developing a processing method for sodium chondroitin sulfate with high extraction rate, fast extraction speed, and high purity is an urgent problem to be solved. Summary of the Invention

[0005] In order to obtain a processing method for chondroitin sulfate sodium with high extraction rate, fast extraction speed and high purity, this application provides a processing technology for chondroitin sulfate sodium.

[0006] This application provides a processing technology for sodium chondroitin sulfate, which adopts the following technical solution: S1. The raw materials are crushed, cooked, alkali-hydrolyzed, and enzymatically hydrolyzed to obtain an enzymatic hydrolysate; S2. The enzymatic hydrolysate is filtered, ultrafiltered, oxidized, filtered again, filtered through an ultrafiltration membrane, precipitated with alcohol, dehydrated, centrifuged, dried, pulverized, and sieved to obtain the finished product.

[0007] By adopting the above technical solution, raw materials such as chicken short bones are pulverized to obtain powder, which is then boiled. The boiling process softens the chicken short bone tissue and destroys the cell structure, making it easier for the proteins and chondroitin sulfate in the cartilage to be released through alkaline and enzymatic hydrolysis. The subsequent alkaline hydrolysis uses alkali to break the covalent bonds between chondroitin sulfate and proteins, releasing chondroitin sulfate and increasing the extraction rate of sodium chondroitin sulfate. The subsequent enzymatic hydrolysis uses enzymes to further hydrolyze and destroy the proteins on the long chains of chondroitin sulfate, forming soluble small molecule peptides, amino acids, and other substances, thus improving the purity of sodium chondroitin sulfate. The combination of alkaline and enzymatic hydrolysis overcomes the excessively long processing time of single alkaline treatment, improving the extraction efficiency of sodium chondroitin sulfate. This ensures that sodium chondroitin sulfate simultaneously possesses the advantages of high extraction rate and high extraction efficiency.

[0008] By employing processes such as filtration, ultrafiltration, oxidation, secondary filtration, and alcohol precipitation, undissolved cartilage fragments and protein precipitates from alkaline and enzymatic hydrolysis are separated. Ultrafiltration can retain large-molecule proteins and polysaccharides, while small-molecule chondroitin sulfate permeates through the membrane pores, thus improving the purity of sodium chondroitin sulfate and reducing residues. Oxidation can improve color and stability, and secondary filtration further refines the product, increasing the purity of sodium chondroitin sulfate and reducing residues.

[0009] Preferably, the cooking process in S1 is as follows: first, heat at 85-95℃ for 10-20 minutes, then heat to 99-100℃ and cook for 30-40 minutes, then cool down to 60-68℃ to obtain a cooking liquid. Add a low DE value maltodextrin complex liquid to the cooking liquid at a mass ratio of 100:5-10 and mix well.

[0010] By adopting the above technical solution, heating at a higher temperature first removes oil and helps soften cartilage tissue, initially disrupting the cross-linking structure between collagen and mucopolysaccharides, allowing chondroitin sulfate to begin to be released from the fibrous network; combined with subsequent heating treatment, the cell wall and cell membrane are further disrupted, significantly increasing cell permeability and accelerating the diffusion of chondroitin sulfate from the cell interior into the extract, thereby greatly improving the extraction rate.

[0011] Then, the temperature is lowered to 60-68℃. Since low DE value maltodextrin is soluble in water above 60℃, and water is gradually lost during the aforementioned heating process, the low DE value maltodextrin complex solution can stably bond with the pulverized raw material powder, forming an adhesion on the surface of the pulverized raw material powder. In the subsequent alkaline hydrolysis and enzymatic hydrolysis processes, it can protect the structural stability of chondroitin sulfate, preventing excessive alkaline hydrolysis that could affect the molecular chain of chondroitin sulfate and ensuring the bioactivity of chondroitin sulfate sodium. Furthermore, the addition amount of low DE value maltodextrin complex solution is limited, allowing the surface of the raw material powder to be loaded with maltodextrin solution without complete blockage. This ensures the penetration of alkaline solution during alkaline hydrolysis and the penetration of enzymes during enzymatic hydrolysis, thereby ensuring the extraction rate and efficiency of chondroitin sulfate sodium while protecting the structural stability of chondroitin sulfate sodium.

[0012] Preferably, the low DE value maltodextrin complex solution is composed of a low DE value maltodextrin solution and ethyl cellulose in a mass ratio of 1:0.1-0.2.

[0013] By adopting the above technical solution, the cooking process softens the tissue and increases its looseness, facilitating contact with alkali and enzymes. However, the outflow of chondroitin sulfate and protein during alkaline and enzymatic hydrolysis can cause shrinkage in the powder structure, affecting the extraction rate and efficiency of sodium chondroitin sulfate. By using a low-DE maltodextrin solution and ethyl cellulose, which are mixed evenly and then applied to the raw material powder, the viscosity of the low-DE maltodextrin solution allows it to adhere to the surface of the powder. Combined with the skeletal support of ethyl cellulose, this further stabilizes the structural stability of the powder during alkaline and enzymatic hydrolysis, preventing excessive shrinkage and ensuring the extraction rate and efficiency of sodium chondroitin sulfate. Furthermore, the low-DE maltodextrin can penetrate and fill the surface pores of the powder, further preventing shrinkage and protecting the bioactivity of sodium chondroitin sulfate, ensuring that the sodium chondroitin sulfate prepared by alkaline and enzymatic hydrolysis has good purity and quality.

[0014] Preferably, in the S1 alkaline hydrolysis process, the material-to-liquid ratio is 1:5-7, the alkaline solution is a 2-4% sodium hydroxide solution, the temperature is 50-56℃, the pH is 9-12, and the hydrolysis time is 1.5-2h.

[0015] By adopting the above technical solution, alkaline conditions (pH 9-12) can effectively disrupt the hydrogen bonds of collagen in chicken short bones, causing collagen fibers to depolymerize and release the encapsulated chondroitin sulfate. The alkaline hydrolysis temperature of 50-56℃ can avoid the degradation of chondroitin sulfate caused by high temperature. Furthermore, excessively high temperature or alkaline concentration can lead to excessive protein denaturation, increasing the difficulty of subsequent purification. Conversely, excessively low temperature or insufficient alkaline concentration will result in incomplete reaction and prolong the extraction time. Therefore, limiting the temperature and time allows for the selective dissolution of chondroitin sulfate. Lipids, non-collagenous proteins, and other impurities have low solubility under alkaline conditions, ensuring the extraction rate, extraction efficiency, and purity of sodium chondroitin sulfate.

[0016] Preferably, the specific steps of the S1 enzymatic hydrolysis are as follows: First, under the conditions of 50-56℃ and pH 7-11, add protease and hydrolyze for 1-1.5h. Then, adjust the pH to 6-8, add neutral protease and amylase, and hydrolyze for 4-6h. Finally, adjust the pH to 5-7, raise the temperature to 60-70℃, and let stand to obtain the enzymatic hydrolysate.

[0017] By employing the above technical solution, enzymatic hydrolysis with protease at 50-56℃ for 1-1.5 hours initially degrades collagen and some non-collagen proteins in cartilage. The initial alkaline treatment breaks down the hydrogen bonds of collagen, making the structure loose and facilitating protease entry to cleave peptide bonds, releasing the encapsulated chondroitin sulfate, and reducing the physical barriers to subsequent enzymatic hydrolysis. Combined with subsequent enzymatic hydrolysis by neutral protease and amylase, the neutral protease efficiently hydrolyzes proteins, and the amylase decomposes maltodextrin, ensuring that the raw material powder can be further decomposed by protease to release chondroitin sulfate, thereby improving the extraction rate and efficiency of chondroitin sulfate sodium. Furthermore, the final acidic conditions inactivate the enzymes, promoting further precipitation of chondroitin sulfate, and the high temperature also destroys enzyme activity, further improving the purity of chondroitin sulfate sodium.

[0018] Preferably, the secondary filtration in S2 uses diatomaceous earth adsorption treatment, and the mass ratio of powder to diatomaceous earth is 9:0.04-0.06.

[0019] By adopting the above technical solution, diatomaceous earth can utilize its porous structure to adsorb incompletely degraded protein fragments, pigments, and small molecule organic matter, thereby improving the purity of sodium chondroitin sulfate.

[0020] Preferably, the diatomaceous earth is prepared by the following method: the diatomaceous earth is crushed, acid-washed, and water-washed. After filtering out the diatomaceous earth, it is soaked in a polyethylene glycol solution. The surface of the filtered diatomaceous earth is uniformly sprayed with chitosan film liquid. The mass ratio of diatomaceous earth to chitosan film liquid is 1:0.8-1.2. The diatomaceous earth is dried at 60-65℃ to obtain the finished product.

[0021] By adopting the above technical solution, chondroitin sulfate is a large-molecule polysaccharide with a molecular weight usually greater than 10 kDa, while small-molecule peptides and free amino acids usually have a molecular weight less than 1 kDa. Therefore, after crushing and acid washing of diatomaceous earth to remove iron and aluminum impurities and expand pores, soaking in polyethylene glycol solution increases the hydroxyl content, promotes the adsorption of small-molecule peptides and amino acids, and improves the purity of chondroitin sulfate sodium. Finally, combined with the coating treatment of chitosan membrane solution, the partial coating effect of chitosan can form a barrier channel. On the one hand, it will not completely block all the pores of diatomaceous earth. On the other hand, the nano-level channels formed by chitosan membrane have mesopores or micropores with a particle size of less than 50 nm, which ensures the penetration of small-molecule peptides and amino acids while blocking the penetration of chondroitin sulfate, thereby removing impurities and improving the purity of chondroitin sulfate sodium.

[0022] Preferably, the chitosan membrane solution is prepared as follows: nano zinc oxide and sodium bicarbonate in a mass ratio of 1:1-2:20-25 are mixed evenly with chitosan solution, and 0.3-0.6% glutaraldehyde by mass is added and mixed evenly to obtain the membrane solution.

[0023] By adopting the above technical solution, a chitosan membrane solution is prepared by combining nano-sized zinc oxide with chitosan solution. The nano-sized zinc oxide is used as a template to induce the chitosan chains to oriented and form nano-sized gaps and channels. Combined with the gas generated by heating sodium bicarbonate, the formation of nano-channels in the chitosan membrane is further promoted. The nano-sized channels, together with the pores of diatomaceous earth, adsorb small molecule peptides and amino acids, and do not easily adsorb sodium chondroitin sulfate, thus ensuring the purity of sodium chondroitin sulfate.

[0024] Preferably, the alcohol precipitation in step S2 is as follows: add ethanol to make the alcohol content in the liquid reach 60-68°, stir for 30-40 minutes, then let stand for 5-16 hours, and remove the supernatant; then add ethanol to adjust the alcohol content to 92-95°, stir for 20-30 minutes, then let stand for 1-2 hours, and remove the supernatant.

[0025] By adopting the above technical solution, sodium chondroitin sulfate is insoluble in water but can form a precipitate in ethanol. First, it is stirred at an alcohol concentration of 60-68° and then allowed to stand to allow the ethanol and chondroitin sulfate to mix evenly and precipitate. Then, it is stirred at an alcohol concentration of 92-95° to gradually dissolve and remove ethyl cellulose. After standing, the ethyl cellulose dissolved in ethanol is in the supernatant, and the chondroitin sulfate precipitates, thus ensuring the purity of sodium chondroitin sulfate.

[0026] Preferably, the specific steps for dehydration in S2 are as follows: Add ethanol solution to adjust the alcohol content of the feed liquid to 70-80°, stir for 1-2 hours, let stand for 0.5-1 hours, remove the supernatant, adjust the alcohol content of the ethanol solution to 82-86°, continue stirring for 1-2 hours, let stand for 0.5-1 hours, remove the supernatant, finally pump in ethanol solution to adjust the alcohol content to 85-90°, continue stirring at 60-65°C for 0.5-1 hours, let stand for 0.5-1 hours, and remove the supernatant.

[0027] By adopting the above technical solution, the temperature is limited during the dehydration process. Under ethanol conditions of 88-92°, combined with stirring, standing time, and repeated operation, the ethyl cellulose is further dissolved. Removing the supernatant removes the ethyl cellulose, ensuring the sedimentation of chondroitin sulfate sodium, thereby further ensuring the purity of chondroitin sulfate sodium.

[0028] In summary, this application has the following beneficial effects: 1. Chicken short bones are crushed and then boiled to soften the bone tissue. This is followed by alkaline hydrolysis, where alkaline substances break the covalent bonds between chondroitin sulfate and protein, releasing the chondroitin sulfate and increasing the extraction rate of sodium chondroitin sulfate. Subsequent enzymatic hydrolysis further hydrolyzes and breaks down the proteins in the long chains of chondroitin sulfate, forming soluble small peptides and amino acids, thus increasing the purity of sodium chondroitin sulfate. This combination of alkaline and enzymatic hydrolysis overcomes the excessively long extraction time of single-alkaline treatment, improving the extraction efficiency of sodium chondroitin sulfate. Impurities are removed and residues are reduced through filtration, ultrafiltration, oxidation, secondary filtration, and alcohol precipitation, further enhancing the purity of sodium chondroitin sulfate.

[0029] 2. Proteases break down collagen, and amylases break down maltodextrin. The addition of a small amount of ethyl cellulose allows it to be removed during subsequent ethanol precipitation, as ethyl cellulose dissolves in ethanol while sodium chondroitin sulfate does not. The added low-DE-value maltodextrin complex not only protects the structural stability of the raw material powder during alkaline hydrolysis, ensuring the extraction rate and efficiency of sodium chondroitin sulfate, but also does not affect its purity. It can be removed during processing, ensuring both the quality and bioactivity of sodium chondroitin sulfate.

[0030] 3. By limiting the amount of ethyl cellulose added to the low DE value maltodextrin compound solution, the ratio of ethyl cellulose to raw materials is controlled. The amount of ethanol added during alcohol precipitation and dehydration, as well as the gradual removal of the supernatant, ensures that ethyl cellulose does not produce excessive viscosity even when dissolved in ethanol. In the final dehydration treatment, ethyl cellulose can be completely removed, ensuring the purity of chondroitin sulfate sodium. Detailed Implementation

[0031] The present application will be further described in detail below with reference to the embodiments.

[0032] All the following ingredients are commercially available food-grade ingredients.

[0033] Preparation Example of Low DE Value Maltodextrin Complex Solution Example 1: The low DE value maltodextrin complex solution was prepared using the following method: Maltodextrin was placed in water and stirred until completely dissolved. The DE value of maltodextrin was 8, and the water temperature was 65℃, resulting in a 3% low DE value maltodextrin solution. 0.15 kg of ethyl cellulose was added to 1 kg of the low DE value maltodextrin solution and mixed evenly to obtain a low DE value maltodextrin composite solution. The average particle size of ethyl cellulose was 3 μm.

[0034] Preparation Example 2: The difference between this preparation example and Preparation Example 1 is that: Maltodextrin was placed in water and stirred until completely dissolved. The DE value of maltodextrin was 8, and the water temperature was 65℃ to obtain a 3% low DE value maltodextrin solution. 0.1 kg of ethyl cellulose was added to 1 kg of low DE value maltodextrin solution and mixed and stirred evenly to obtain a low DE value maltodextrin composite solution.

[0035] Preparation Example 3: The difference between this preparation example and Preparation Example 1 is that: Maltodextrin was placed in water and stirred until completely dissolved. The DE value of maltodextrin was 8, and the water temperature was 65℃ to obtain a 3% low DE value maltodextrin solution. 0.2 kg of ethyl cellulose was added to 1 kg of low DE value maltodextrin solution and mixed and stirred evenly to obtain a low DE value maltodextrin composite solution.

[0036] Preparation Example 4: Diatomaceous earth was prepared using the following method: 1 kg of nano zinc oxide, 1.5 kg of sodium bicarbonate and 22.5 kg of chitosan solution with a mass fraction of 1.5% were mixed evenly. The average particle size of the nano zinc oxide was 40 nm and the average particle size of the sodium bicarbonate was 80 nm. The solvent of the chitosan solution was an aqueous acetic acid solution with a mass fraction of 2%. The degree of deacetylation of the chitosan was 85%. 0.5% glutaraldehyde was added and stirred for 20 min. After mixing evenly, the chitosan film solution was obtained. Diatomaceous earth is crushed to 250 mesh, then soaked in 10% citric acid for 10 minutes, followed by water washing for 5 minutes. After filtering out the diatomaceous earth, it is soaked in a polyethylene glycol solution with a mass ratio of 1:8 (diatomaceous earth to polyethylene glycol solution, 5% polyethylene glycol aqueous solution, polyethylene glycol 1000). After soaking and contact, the diatomaceous earth is filtered out. Then, chitosan film solution is evenly sprayed onto the surface with a mass ratio of 1:1 (diatomaceous earth to chitosan film solution). It is then air-dried at 65℃ to remove moisture from the polyethylene glycol solution while the chitosan film solution dries to form a film, yielding the finished product. The finished product passes through a 150-mesh sieve.

[0037] Preparation Example 5: The difference between this preparation example and Preparation Example 4 is that: Mix 1 kg of nano zinc oxide, 1 kg of sodium bicarbonate and 20 kg of 1% chitosan solution evenly, add 0.3% glutaraldehyde, stir for 20 min, and after mixing evenly, obtain chitosan film solution. Diatomaceous earth is crushed to 250 mesh, then soaked in 10% citric acid for 10 minutes, followed by water washing for 5 minutes. After filtering out the diatomaceous earth, it is soaked in polyethylene glycol solution at a mass ratio of 1:8. Then, chitosan film solution is evenly sprayed onto the surface at a mass ratio of 1:0.8. The mixture is dried at 60°C to obtain the finished product, which is then passed through a 150-mesh sieve.

[0038] Preparation Example 6: The difference between this preparation example and Preparation Example 4 is that: 1 kg of nano zinc oxide, 2 kg of sodium bicarbonate and 25 kg of chitosan solution with a mass fraction of 2% were mixed evenly, 0.6% glutaraldehyde was added, and the mixture was stirred for 20 min. After mixing evenly, chitosan film solution was obtained. Diatomaceous earth is crushed to 250 mesh, then soaked in 10% citric acid for 10 minutes, followed by water washing for 5 minutes. After filtering out the diatomaceous earth, it is soaked in polyethylene glycol solution at a mass ratio of 1:8. Then, chitosan film solution is evenly sprayed onto the surface at a mass ratio of 1:1.2. The mixture is dried at 65°C to obtain the finished product, which is then passed through a 150-mesh sieve. Example

[0039] Example 1: A processing method for sodium chondroitin sulfate: S1. The raw material is chicken short bones. After thawing frozen chicken short bones, 9 tons per tank are pulverized and passed through a 40-mesh sieve to obtain powder. This powder is then transferred to a cooking tank with a material-to-liquid ratio of 1:5. The steam valve is opened, and the mixture is first heated at 90℃ for 15 minutes, then heated to 100℃ for 35 minutes, and then cooled to 65℃ to obtain a cooking liquid. The low DE value maltodextrin complex liquid prepared in Example 1 is added to the cooking liquid at a mass ratio of 100:8. The mixture is stirred evenly, allowed to stand, and then the oil is skimmed off to complete the cooking process. After cooking, alkaline hydrolysis is performed with a material-to-liquid ratio of 1:6. The alkaline solution is a 3% sodium hydroxide solution, the solvent is water, the temperature is 54℃, the pH is adjusted to 10, and the hydrolysis time is 2 hours. Alkaline hydrolysis was completed in h; then enzymatic hydrolysis was performed. First, papain (200,000 enzyme activity (u / g)) was added at 55℃ and pH 10, with a mass ratio of powder to papain of 3:0.002. Enzymatic hydrolysis was carried out for 1 h. Then, the pH was adjusted to 7, and neutral protease (65,000 enzyme activity (u / g)) and amylase were added. The amylase was thermostable α-amylase (20,000 enzyme activity (u / g)), with a mass ratio of powder to neutral protease and amylase of 9:0.002:0.0005. Enzymatic hydrolysis was carried out for 5 h, with temperature and pH measured every 1 h. The conditions were maintained. Finally, the pH was adjusted to 6, the temperature was raised to 65℃, and the mixture was allowed to stand for 2 h to obtain the enzymatic hydrolysate. The enzymatic hydrolysis treatment was completed, and the enzymatic hydrolysate was obtained. S2. Place the enzymatic hydrolysate in a filter tank, add 200-mesh commercially available diatomaceous earth (powder to diatomaceous earth mass ratio 9:0.035), turn on the filter, and circulate for 20 minutes to complete the filtration process. Then, control the feed temperature at 60℃ and perform ultrafiltration at a post-membrane pressure of 0.6MPa to concentrate the feed to 6m³. 3 Add 8m 3The feed solution was diluted with RO water, then concentrated again. This process of dilution and concentration was repeated three times. Finally, the feed solution in the equipment was replaced with RO water and mixed with the concentrated feed solution, then transferred to the oxidation tank. Stirring was started, and sodium chloride was added to the oxidation tank at a powder-to-sodium chloride mass ratio of 3:0.1. The feed solution temperature was controlled at 55℃. Sodium hydroxide was added to adjust the pH to 11, and hydrogen peroxide was added at a powder-to-hydrogen peroxide mass ratio of 3:0.01. The temperature was maintained at 55℃, and the reaction was carried out for 4 hours, with the temperature measured every hour to complete the oxidation process. After oxidation, the feed solution was heated to 64℃, and diatomaceous earth prepared in Example 4 was added at a powder-to-diatomaceous earth mass ratio of 9:0.05. The filter was started, and the circulation time was 20 minutes to complete the secondary filtration. The filtered feed solution was then filtered through an ultrafiltration membrane, and the pre-membrane pressure was controlled at 0.5 MPa for concentration. The final total volume of the feed solution was 3.3 m³. 3 The ultrafiltration membrane filtration is completed. After adjusting the pH of the feed solution to 6 with hydrochloric acid, it is transferred to an alcohol precipitation tank. Stirring is started, and 99% ethanol is pumped in to bring the alcohol content to 65°. Stirring is continued for 35 minutes, followed by 12 hours of sedimentation. The supernatant is then removed, and 99% ethanol is added to adjust the alcohol content to 94°. Stirring is continued for 25 minutes, followed by 1.5 hours of sedimentation. The supernatant is then removed, completing the alcohol precipitation process. A 90° ethanol solution is prepared in an ethanol preparation tank. This 90° ethanol solution is pumped into the material after the supernatant has been removed during sedimentation, adjusting the alcohol content to 75°. Stirring is continued at room temperature for 1.5 hours, followed by 1 hour of sedimentation. The supernatant is then removed, and the 90° ethanol solution is pumped in to adjust the alcohol content to 84°. Stirring continues at room temperature for 1.5 hours, followed by 1 hour of sedimentation. The supernatant is then removed. The supernatant is then pumped into ethanol to adjust the alcohol content to 88°. Stirring continues for 1 hour at 62°C, followed by 1 hour of settling. The supernatant is then removed, and the material is ready for discharge, completing the dehydration process. Ethanol is continuously added during this process to maintain the alcohol content. The ethanol used is then distilled to remove impurities and increase the ethanol concentration. The distilled ethanol is recovered, stored, and reused. The discharged wet material is centrifuged, not exceeding 2 / 3 of the centrifuge volume, for 5 minutes each time. It is then dried under a vacuum of -0.07 MPa and at 60°C for 4 hours to complete the drying process. The material is then poured into a ball mill for pulverization. The pulverized semi-finished product is then passed through a magnetic rod to remove iron impurities, sieved through an 80-mesh sieve, packaged in polyethylene bags, labeled, and stored in a light-proof environment to obtain the finished product.

[0040] Example 2: The difference between this example and Example 1 is that: S1. The raw material is chicken short bones. After thawing frozen chicken short bones, 9 tons per tank are pulverized and passed through a 40-mesh sieve to obtain powder. This powder is then transferred to a cooking tank with a material-to-liquid ratio of 1:5. The steam valve is opened, and the mixture is first heated at 85°C for 20 minutes, then heated to 99°C for 40 minutes, and then cooled to 60°C to obtain a cooking liquid. The low DE value maltodextrin complex liquid prepared in Preparation Example 2 is added to the cooking liquid at a mass ratio of 100:5. The mixture is stirred evenly, allowed to stand, and then the oil is skimmed off to complete the cooking process. After cooking, alkaline hydrolysis is performed with a material-to-liquid ratio of 1:5. The alkaline solution is a 2% sodium hydroxide solution, the solvent is water, the temperature is 50°C, the pH is adjusted to 9, and the hydrolysis time is 1.5 minutes. Alkaline hydrolysis was completed in h; then enzymatic hydrolysis was performed. First, papain (200,000 enzyme activity (u / g)) was added at 50℃ and pH 7, with a mass ratio of powder to papain of 3:0.002. Enzymatic hydrolysis was performed for 1 h. Then, the pH was adjusted to 6, and neutral protease (65,000 enzyme activity (u / g)) and amylase were added. The amylase was thermostable α-amylase (20,000 enzyme activity (u / g)), with a mass ratio of powder to neutral protease and amylase of 9:0.002:0.0005. Enzymatic hydrolysis was performed for 4 h, with temperature and pH measured every 1 h. The conditions were maintained. Finally, the pH was adjusted to 5, the temperature was raised to 70℃, and the mixture was allowed to stand for 2 h to obtain the enzymatic hydrolysate. The enzymatic hydrolysis treatment was completed, and the enzymatic hydrolysate was obtained. S2. The enzymatic hydrolysate was placed in a filter tank, and diatomaceous earth was added at a powder-to-diatomaceous earth mass ratio of 9:0.035. The filter was turned on and circulated for 20 minutes to complete the filtration process. Then, the feed temperature was controlled at 60℃, and ultrafiltration was performed under a post-membrane pressure of 0.6MPa to concentrate the feed to 6m³. 3 Add 8m 3 The feed solution was diluted with RO water, and then further concentrated. This process of dilution and concentration was repeated three times. Finally, the feed solution in the equipment was replaced with RO water, mixed with the concentrated feed solution, and transferred to the oxidation tank. Stirring was started, and sodium chloride was added to the oxidation tank at a powder-to-sodium chloride mass ratio of 3:0.1. The feed solution temperature was controlled at 50℃. Sodium hydroxide was added to adjust the pH to 10, and hydrogen peroxide was added at a powder-to-hydrogen peroxide mass ratio of 3:0.01. The temperature was maintained at 50℃, and the reaction was carried out for 3 hours. The temperature was measured every hour to complete the oxidation process. After oxidation, the feed solution was heated to 60℃, and diatomaceous earth prepared in Example 5 was added at a powder-to-diatomaceous earth mass ratio of 9:0.04. The filter was started, and the circulation time was 20 minutes to complete the secondary filtration. The filtered feed solution was then filtered through an ultrafiltration membrane, and the pre-membrane pressure was controlled at 0.5 MPa for concentration. The final total volume of the feed solution was 3.3 m³. 3The ultrafiltration membrane filtration is completed. After adjusting the pH of the feed solution to 6 with hydrochloric acid, it is transferred to an alcohol precipitation tank. Stirring is started, and 99% ethanol is pumped in to bring the alcohol content of the feed solution to 60°. Stirring is performed for 30 minutes, and then the solution is allowed to stand for 5 hours for alcohol precipitation. The supernatant is removed, and then 99% ethanol is added to adjust the alcohol content to 92°. Stirring is performed for 30 minutes, and then the solution is allowed to stand for 1 hour. The supernatant is removed to complete the alcohol precipitation process. An 88° ethanol solution is prepared in an ethanol preparation tank. The 88° ethanol solution is pumped into the material after the supernatant has been removed from the alcohol precipitation tank to adjust the alcohol content of the feed solution to 70°. Stirring is performed for 1 hour, and the solution is allowed to stand for 0.5 hours. The supernatant is then removed, and the 88° ethanol solution is pumped in to adjust the alcohol content to 82°. Stirring is performed for 1 hour, and the solution is allowed to stand for 0.5 hours. The supernatant is then removed, and finally, ethanol is pumped in. The alcohol concentration was adjusted to 85°, and the mixture was stirred at 60°C for 1 hour. After standing for 0.5 hours, the supernatant was removed, and the mixture was ready to be discharged to complete the dehydration process. Ethanol was continuously added during this process to maintain the alcohol concentration. The ethanol used was distilled to remove impurities and increase the ethanol concentration. The distilled ethanol was recovered, stored, and reused. The discharged wet material was centrifuged, with the centrifuge volume not exceeding 2 / 3, for 5 minutes each time. Then, it was dried under a vacuum of -0.07 MPa and a temperature of 60°C for 4 hours to complete the drying process. The material was then poured into a ball mill for pulverization. The pulverized semi-finished product was then passed through a magnetic rod to remove iron impurities, sieved through an 80-mesh sieve, packaged in polyethylene bags, labeled, and stored in the dark to obtain the finished product.

[0041] Example 3: The difference between this example and Example 1 is that: S1. The raw material is chicken short bones. After thawing frozen chicken short bones, 9 tons per tank are pulverized and passed through a 40-mesh sieve to obtain powder. This powder is then transferred to a cooking tank with a material-to-liquid ratio of 1:5. The steam valve is opened, and the mixture is first heated at 95°C for 10 minutes, then heated to 100°C for 30 minutes, and then cooled to 68°C to obtain a cooking liquid. The low DE value maltodextrin complex liquid prepared in Preparation Example 3 is added to the cooking liquid at a mass ratio of 100:10. The mixture is stirred evenly, allowed to stand, and then the oil is skimmed off to complete the cooking process. After cooking, alkaline hydrolysis is performed with a material-to-liquid ratio of 1:7. The alkaline solution is a 4% sodium hydroxide solution, the solvent is water, the temperature is 56°C, the pH is adjusted to 12, and the hydrolysis time is 1.5 minutes. Alkaline hydrolysis was completed in h; then enzymatic hydrolysis was performed. First, papain (200,000 enzyme activity (u / g)) was added at 56℃ and pH 11, with a mass ratio of powder to papain of 3:0.002, and enzymatic hydrolysis was carried out for 1.5 h. Then, the pH was adjusted to 8, and neutral protease (65,000 enzyme activity (u / g)) and amylase were added. The amylase was thermostable α-amylase (20,000 enzyme activity (u / g)), with a mass ratio of powder to neutral protease and amylase of 9:0.002:0.0005, and enzymatic hydrolysis was carried out for 6 h. The temperature and pH were measured every 1 h to maintain the conditions. Finally, the pH was adjusted to 7, the temperature was raised to 60℃, and the mixture was allowed to stand for 2 h to obtain the enzymatic hydrolysate. The enzymatic hydrolysis treatment was completed, and the enzymatic hydrolysate was obtained. S2. The enzymatic hydrolysate was placed in a filter tank, and diatomaceous earth was added at a powder-to-diatomaceous earth mass ratio of 9:0.035. The filter was turned on and circulated for 20 minutes to complete the filtration process. Then, the feed temperature was controlled at 60℃, and ultrafiltration was performed under a post-membrane pressure of 0.6MPa to concentrate the feed to 6m³. 3 Add 8m 3 The feed solution was diluted with RO water, then concentrated again. This process of dilution and concentration was repeated three times. Finally, the feed solution in the equipment was replaced with RO water and mixed with the concentrated feed solution, then transferred to the oxidation tank. Stirring was started, and sodium chloride was added to the oxidation tank at a powder-to-sodium chloride mass ratio of 3:0.1. The feed solution temperature was controlled at 56℃. Sodium hydroxide was added to adjust the pH to 12, and hydrogen peroxide was added at a powder-to-hydrogen peroxide mass ratio of 9:0.04. The temperature was maintained at 56℃, and the reaction was carried out for 5 hours. The temperature was measured every hour to complete the oxidation process. After oxidation, the feed solution was heated to 64℃, and diatomaceous earth prepared in Example 6 was added at a powder-to-diatomaceous earth mass ratio of 9:0.05. The filter was started, and the circulation time was 20 minutes to complete the secondary filtration. The filtered feed solution was then filtered through an ultrafiltration membrane, and the pre-membrane pressure was controlled at 0.5 MPa for concentration. The final total volume of the feed solution was 3.3 m³. 3 The ultrafiltration membrane filtration is completed. After adjusting the pH of the feed solution to 6 with hydrochloric acid, it is transferred to an alcohol precipitation tank. Stirring is started, and ethanol is pumped in to bring the alcohol content of the feed solution to 68°. Stirring is continued for 40 minutes, followed by standing for 16 hours of alcohol precipitation. The supernatant is then removed, and ethanol is added again to adjust the alcohol content to 95°. Stirring is continued for 20 minutes, followed by standing for 2 hours. The supernatant is then removed to complete the alcohol precipitation process. A 90° ethanol solution is prepared in an ethanol preparation tank. This 90° ethanol solution is pumped into the material after the supernatant has been removed from the alcohol precipitation tank to adjust the alcohol content to 80°. The mixture is stirred at 68° for 2 hours, allowed to stand for 1 hour, and then the supernatant is removed. The 90° ethanol solution is then pumped in to adjust the alcohol content to 86°. Stirring is continued for 2 hours, allowed to stand for 1 hour, and then the supernatant is removed. The supernatant is then pumped into ethanol to adjust the alcohol content to 90°, and stirring is continued for 2 hours. After standing for 1 hour, the supernatant is pumped out and the material is ready to be discharged, completing the dehydration process. The ethanol used is then distilled to remove impurities and increase the ethanol concentration. The distilled ethanol is recovered, stored, and reused. The discharged wet material is centrifuged, not exceeding 2 / 3 of the centrifuge volume, for 5 minutes each time. Then, it is dried under a vacuum of -0.07 MPa and a temperature of 60°C for 4 hours to complete the drying process. The material is then poured into a ball mill for pulverization. The pulverized semi-finished product is then passed through a magnetic rod to remove iron impurities, sieved through an 80-mesh sieve, packaged in polyethylene bags, labeled, and stored in the dark to obtain the finished product.

[0042] Example 4: The difference between this example and Example 1 is that: No low-DE value maltodextrin complex was added during the cooking process of S1.

[0043] Example 5: The difference between this example and Example 1 is that: Ethyl cellulose was not added during the preparation of the S1 low DE value maltodextrin complex.

[0044] Example 6: The difference between this example and Example 1 is that: In the S2 secondary filtration process, no polyethylene glycol solution was added for soaking during the preparation of diatomaceous earth.

[0045] Example 7: The difference between this example and Example 1 is that: In the S2 secondary filtration process, no chitosan membrane liquid was added during the preparation of diatomaceous earth.

[0046] Example 8: The difference between this example and Example 1 is that: No nano zinc oxide or sodium bicarbonate was added during the preparation of the chitosan membrane solution in the diatomaceous earth of the S2 secondary filtration process.

[0047] Example 9: The difference between this example and Example 1 is that: The specific steps for S2 alcohol precipitation and dehydration are as follows: Start the stirring, pump in ethanol to bring the alcohol content of the liquid to 65°, stir for 35 minutes, then let it stand for 12 hours for alcohol precipitation, remove the supernatant to complete the alcohol precipitation process; prepare 90° ethanol solution in the ethanol preparation tank, pump in the 90° ethanol solution to the material from which the supernatant has been removed during alcohol precipitation, adjust the alcohol content of the solution to 75°, stir at room temperature for 1.5 hours, let it stand for 1 hour, then remove the supernatant, pump in the 90° ethanol solution to adjust the alcohol content to 84°, continue stirring at room temperature for 1.5 hours, let it stand for 1 hour, remove the supernatant, and prepare to discharge the material to complete the dehydration process.

[0048] Comparative Example Comparative Example 1: The difference between this comparative example and Example 1 is that: It has not undergone enzymatic hydrolysis.

[0049] Comparative Example 2: This comparative example differs from Example 1 in that: It has not undergone secondary filtration or ultrafiltration membrane filtration.

[0050] Performance testing 1. Extraction rate detection Chondroitin sulfate sodium was prepared using the methods of Examples 1-8 and Comparative Example 1, respectively. The content of chondroitin sulfate sodium was detected by high performance liquid chromatography. The extraction rate was calculated as: (Chondroitin sulfate sodium content in finished product / Theoretical chondroitin sulfate sodium content in raw material) × 100%. The data were recorded.

[0051] 2. Purity testing Chondroitin sulfate sodium was prepared using the methods of Examples 1-3, 6-9 and Comparative Example 2, respectively. The purity was calculated and the data were recorded using high performance liquid chromatography.

[0052] Table 1 Performance Test Table (In the table below, " / " indicates that this item was not tested and no data is displayed) As can be seen from Examples 1-3 and Table 1, the chondroitin sulfate sodium prepared in this application has a good extraction rate and high purity.

[0053] Combining Examples 1 and 4-9 with Table 1, it can be seen that in Example 4, no low DE value maltodextrin complex solution was added during the cooking process. Compared with Example 1, the extraction rate of Example 4 was lower than that of Example 1. This indicates that the binding effect of the low DE value maltodextrin complex solution can protect the structural stability of chondroitin sulfate during subsequent alkaline and enzymatic hydrolysis, preventing excessive alkaline hydrolysis that could affect the molecular chain of chondroitin sulfate. It also ensures the penetration of alkaline solution during alkaline hydrolysis and the penetration of enzymes during enzymatic hydrolysis, thereby ensuring the extraction rate and efficiency of chondroitin sulfate sodium while protecting the structural stability of chondroitin sulfate sodium, thus ensuring the extraction outflow of chondroitin sulfate sodium and maintaining the extraction rate.

[0054] In Example 5, no ethyl cellulose was added during the preparation of the low DE value maltodextrin complex solution. Compared with Example 1, the extraction rate of Example 5 was lower than that of Example 1. This indicates that ethyl cellulose has a supporting effect, which can further support the structural stability of the raw material powder during the extraction process, ensure the extraction of chondroitin sulfate sodium, and ensure the extraction rate.

[0055] In Example 6, no polyethylene glycol solution was added during the diatomaceous earth preparation process in the secondary filtration. Compared with Example 1, the extraction rate and purity of Example 6 were lower than those of Example 1. This indicates that the polyethylene glycol solution can adsorb small molecule peptides and amino acids, promote the adsorption of impurities by diatomaceous earth during the secondary filtration process, and improve the purity of sodium chondroitin sulfate.

[0056] In Example 7, no chitosan membrane solution was added during the secondary filtration process of diatomaceous earth preparation. Compared with Example 1, the extraction rate and purity of Example 7 were lower than those of Example 1. This indicates that the treatment with chitosan membrane solution can further promote the adsorption of impurities by diatomaceous earth and improve the purity of sodium chondroitin sulfate.

[0057] In Example 8, no nano-zinc oxide and sodium bicarbonate were added during the preparation of the chitosan membrane solution in the diatomaceous earth after secondary filtration. Compared with Example 1, the extraction rate and purity of Example 8 were lower than those of Example 1. This indicates that the addition of nano-zinc oxide and sodium bicarbonate can create microporous channels, promote the permeation of small molecule peptides and amino acids, block the permeation of chondroitin sulfate, control the adsorption of chondroitin sulfate by diatomaceous earth, and ensure the extraction rate and purity of chondroitin sulfate sodium.

[0058] Compared to Example 1, the purity of Example 9 is lower. This indicates that ethyl cellulose can gradually dissolve during the alcohol precipitation process at an alcohol content of 94°. Combined with the conditions of dehydration at an alcohol content of 88° and continued stirring at 62° for 1 hour, the solubility of ethyl cellulose can be further improved. The gradually removed supernatant and the amount of ethyl cellulose added control the viscosity of ethyl cellulose dissolved in ethanol, ensuring the removal of ethyl cellulose while maintaining the purity of chondroitin sulfate sodium.

[0059] Based on Example 1 and Comparative Examples 1-2, and in conjunction with Table 1, it can be seen that Comparative Example 1, which was not subjected to enzymatic hydrolysis, had a lower extraction rate than Example 1. This indicates that enzymatic hydrolysis-assisted alkaline hydrolysis can improve the extraction rate.

[0060] Comparative Example 2 did not undergo secondary filtration or ultrafiltration. Compared to Example 1, the purity of Comparative Example 2 was lower than that of Example 1. This indicates that secondary filtration and ultrafiltration can remove impurities and improve purity.

[0061] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A processing method for sodium chondroitin sulfate, characterized in that, Includes the following steps: S1. The raw materials are crushed, cooked, alkali-hydrolyzed, and enzymatically hydrolyzed to obtain an enzymatic hydrolysate; S2. The enzymatic hydrolysate is filtered, ultrafiltered, oxidized, filtered again, filtered through an ultrafiltration membrane, precipitated with alcohol, dehydrated, centrifuged, dried, pulverized, and sieved to obtain the finished product.

2. The processing method for sodium chondroitin sulfate according to claim 1, characterized in that: The specific steps of cooking in S1 are as follows: first, heat at 85-95℃ for 10-20 minutes, then raise the temperature to 99-100℃ and cook for 30-40 minutes, then lower the temperature to 60-68℃ to obtain the cooking liquid. Add low DE value maltodextrin complex liquid to the cooking liquid at a mass ratio of 100:5-10 and mix well.

3. The processing method for sodium chondroitin sulfate according to claim 2, characterized in that, The low DE value maltodextrin complex solution is composed of a low DE value maltodextrin solution and ethyl cellulose in a mass ratio of 1:0.1-0.

2.

4. The processing method for sodium chondroitin sulfate according to claim 1, characterized in that, During the S1 alkaline hydrolysis process, the material-to-liquid ratio is 1:5-7, the alkaline solution is a 2-4% sodium hydroxide solution, the temperature is 50-56℃, the pH is 9-12, and the hydrolysis time is 1.5-2h.

5. The processing method for sodium chondroitin sulfate according to claim 1, characterized in that, The specific steps of S1 enzymatic hydrolysis are as follows: First, under the conditions of 50-56℃ and pH 7-11, add protease and hydrolyze for 1-1.5h. Then, adjust the pH to 6-8, add neutral protease and amylase, and hydrolyze for 4-6h. Finally, adjust the pH to 5-7, raise the temperature to 60-70℃, and let it stand to obtain the enzymatic hydrolysate.

6. The processing method for sodium chondroitin sulfate according to claim 1, characterized in that, In the S2 process, the secondary filtration is performed using diatomaceous earth adsorption, with the mass ratio of powder to diatomaceous earth being 9:0.04-0.

06.

7. The processing method for sodium chondroitin sulfate according to claim 6, characterized in that, The diatomaceous earth is prepared by the following method: the diatomaceous earth is crushed, acid-washed, and water-washed. After filtering out the diatomaceous earth, it is soaked in a polyethylene glycol solution. The surface of the filtered diatomaceous earth is uniformly sprayed with chitosan film solution. The mass ratio of diatomaceous earth to chitosan film solution is 1:0.8-1.

2. The diatomaceous earth is dried at 60-65℃ to obtain the finished product.

8. The processing method for sodium chondroitin sulfate according to claim 7, characterized in that, The chitosan membrane solution is prepared as follows: nano zinc oxide and sodium bicarbonate in a mass ratio of 1:1-2:20-25 are mixed evenly with chitosan solution, and 0.3-0.6% glutaraldehyde by mass is added and mixed evenly to obtain the membrane solution.

9. The processing method for sodium chondroitin sulfate according to claim 1, characterized in that, The specific steps of alcohol precipitation in S2 are as follows: add ethanol to make the alcohol content in the liquid reach 60-68°, stir for 30-40 minutes, then let it stand for 5-16 hours, and remove the supernatant; then add ethanol to adjust the alcohol content to 92-95°, stir for 20-30 minutes, then let it stand for 1-2 hours, and remove the supernatant.

10. The processing method for chondroitin sulfate sodium according to claim 1, characterized in that, The specific steps for dehydration in S2 are as follows: Add ethanol to adjust the alcohol content of the feed solution to 70-80°, stir for 1-2 hours, let stand for 0.5-1 hours, remove the supernatant, adjust the alcohol content of the ethanol to 82-86°, continue stirring for 1-2 hours, let stand for 0.5-1 hours, remove the supernatant, and finally pump in ethanol to adjust the alcohol content to 85-90°. Continue stirring at 60-65° for 0.5-1 hours, let stand for 0.5-1 hours, and remove the supernatant.

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