CN111533826A - Method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones - Google Patents
Method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones Download PDFInfo
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Abstract
The invention discloses a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones, which adopts a new double-enzyme process, removes the steps of alkali extraction and oxidation, simultaneously obtains a large amount of high-quality collagen, reduces the material consumption, simplifies the operation, realizes a new co-production process of the chondroitin sulfate and the collagen peptide, realizes the clean production of the chondroitin sulfate, avoids the problem of water pollution caused by protein discharge in the traditional production process, and is particularly suitable for popularization and application of industrial production.
Description
Technical Field
The invention relates to clean production of chondroitin sulfate, in particular to a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones. Belongs to the technical field of pig bone extraction.
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.
Most of the existing production processes of chondroitin sulfate are alkaline hydrolysis, then the chondroitin sulfate is subjected to enzymolysis by pancreas and alkaline protease, and the chondroitin sulfate is released from proteoglycan. However, in the alkali extraction process, the color of the extract is deepened, the later protein removal is difficult, the color and the quality of the product are seriously influenced, and oxidation and decoloration treatment needs to be carried out in the subsequent process. Therefore, the prior art has the defects of long production period, high production cost, unstable product quality and the like. Meanwhile, a large amount of protein impurities are generated in the production, and the environment is seriously polluted.
In addition, pure cartilage or fresh bone containing partial cartilage is often selected for production in the current production, the precious resources are rarely utilized in chondroitin sulfate extraction factories except for extracting chondroitin, and most of water-soluble protein is discharged as wastewater, so that the serious pollution problem is also brought. Therefore, the deep development of fresh bone resources and the effective improvement of the comprehensive utilization value thereof are very important and significant works.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones, thereby not only realizing the maximization of economic value, but also solving the problem of protein pollution.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, performing first extraction for 60-80 minutes at 115-120 ℃, and then performing second extraction for 50-60 minutes at 130-135 ℃ to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting collagen peptides with a molecular weight of less than 1000Da (weight average) by the ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
Preferably, in the step (1), the pig bones are firstly crushed and then added into a first part of water with the weight of 2-3 times, and first extraction and filtration are carried out to obtain a first extraction liquid and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, performing secondary extraction, filtering to obtain a second extract, and combining the second extract with the first extract to obtain the extract.
Further preferably, the pig bone is pulverized as follows: crushing fresh and clean pig bones to 3-5 mm by using a powerful osteoclast machine, then adding the crushed pig bones into water with the weight of 1-1.5 times of that of the pig bones, then grinding the pig bones for 5-8 minutes by a wet method, centrifuging, drying and scattering the ground pig bones to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m.
Further preferably, the wet grinding is carried out by a colloid mill, the gap between two grinding discs of the colloid mill is 0.02-0.03 mm, and the rotating speed is 10000-12000 r/min.
Still more preferably, the process conditions of centrifugation are: centrifuging at 10000-12000 r/min for 5-6 minutes, and discarding the supernatant.
Further preferably, the drying process is carried out at 90-100 ℃ for 5-8 hours.
It is further preferred that the dried material is broken up by means of a gas stream.
Preferably, in step (2), before the first enzymolysis, the floating oil on the surface of the extract is skimmed.
Preferably, in the step (2), the process conditions of the first enzymolysis are as follows: adding a first complex enzyme with the weight of 0.004-0.006 time of the weight of the pig bones into the extract, and carrying out enzymolysis treatment for 2 hours at the pH of 8.5 and the temperature of 54 ℃.
Further preferably, the first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease and 0.3 to 0.5 part of trypsin.
In the step (2), the resin adsorption time is 3-5 hours, the diameter-height ratio of the used resin column is 1: 3-15, and the resin type is as follows: rohm and Hass (AMBERLITE) acrylic strong base anion exchange resin FPA94CL (food grade) with particle size of 0.630-0.850 mm.
Preferably, in the step (2), the process conditions of the second enzymolysis are as follows: adding a second complex enzyme with the weight 0.004-0.006 time of that of the pig bone, and carrying out enzymolysis treatment for 2 hours at the pH of 7.0 and the temperature of 53 ℃.
Further preferably, the second complex enzyme comprises: 1 part of keratinase, 0.4-0.6 part of bromelain and 0.2-0.3 part of neutral protease.
Preferably, in the step (2), the process conditions of the secondary shearing are as follows: the first shearing is carried out at 6000-8000 rpm for 30-60 minutes, and the first shearing is carried out at 2000-3000 rpm for 80-120 minutes.
Preferably, in the step (4), the specific method of oxidation is: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
Preferably, in the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity and the concentration ratio of the permeate liquid reach the required range, preparing to convey the concentrated feed liquid into the oxidation tank, and simultaneously checking whether the switching of the valve in the oxidation tank is correct.
Further preferably, the salinity requirement range of the permeating liquid is determined according to the actual condition and is usually 5-6%; the concentration ratio is usually controlled to be 1: 3-4, i.e. the concentration is 1/3-1/4 of the original volume.
Preferably, in the step (4), the alcohol precipitation method specifically comprises: adding 70-80% ethanol water solution with volume concentration equal to the volume concentration of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
Preferably, in the step (4), the drying process conditions are as follows: drying the mixture for 8 to 12 hours at the temperature of between 55 and 60 ℃.
The invention has the beneficial effects that:
the invention adopts a new double-enzyme process, removes the steps of alkali extraction and oxidation, simultaneously obtains a large amount of high-quality collagen, reduces the material consumption, simplifies the operation, realizes the new process for co-producing chondroitin sulfate and collagen peptide, realizes the clean production of chondroitin sulfate, also avoids the problem of water pollution caused by protein discharge in the traditional production process, and is particularly suitable for popularization and application of industrial production.
The temperature of the secondary extraction is low before and high after, which is more beneficial to the free treatment of chondroitin sulfate and protein and improves the yield of the chondroitin sulfate and the collagen peptide. The enzymolysis is divided into two times, the first time of enzymolysis is carried out under the condition of higher pH by utilizing the combination of alkaline protease and trypsin, and the second time of enzymolysis is carried out under the condition of lower pH by utilizing keratinase, bromelain and neutral protease, so that the collagen can be more fully degraded, and a small peptide product can be obtained. The elution liquid obtained after the resin adsorbs the chondroitin sulfate is desalted and concentrated by using a hollow fiber membrane, so that the dosage of ethanol in the step of alcohol precipitation is greatly reduced.
The invention also carries out crushing treatment on the pig bones in advance during secondary extraction, and the specific method is that firstly, a powerful osteoclast is utilized to crush the pig bones to a millimeter level, then, the pig bones are ground into a micron level by a wet method, fine pig bone particles can release chondroitin sulfate and ossein protein more easily in the extraction process, and the extraction rate is improved.
The inventive process is compared to the conventional process as shown in table 1.
TABLE 1 comparison of the Processes
Detailed Description
The present invention will be further illustrated by the following examples, which are intended to be merely illustrative and not limitative.
Example 1:
a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, firstly performing first extraction for 60 minutes at 115 ℃, and then performing second extraction for 50 minutes at 130 ℃ to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting ossein peptide with molecular weight lower than 1000Da through ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
In the step (1), crushing pig bones, adding the crushed pig bones into a first part of water with the weight 2 times that of the crushed pig bones, performing first extraction, and filtering to obtain a first extraction solution and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, carrying out secondary extraction, filtering to obtain a second extraction solution, and combining the second extraction solution with the first extraction solution to obtain the extraction solution.
The pig bone crushing method comprises the following steps: crushing fresh and clean pig bones to 3mm by using a powerful osteoclast machine, then adding the crushed pig bones into 1 time of water by weight, then grinding the pig bones for 5 minutes by a wet method, centrifuging, drying and scattering the pig bones to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m. And wet grinding is carried out by adopting a colloid mill, the gap between two grinding discs of the colloid mill is 0.02mm, and the rotating speed is 10000 r/min. The process conditions of centrifugation are as follows: centrifuging at 10000r/min for 5 min, and discarding the supernatant. The drying process condition is drying for 5 hours at 90 ℃. And scattering the dried material by using air flow.
In the step (2), before the first enzymolysis, floating oil on the surface of the extract is skimmed.
In the step (2), the process conditions of the first enzymolysis are as follows: adding 0.004 times of pig bone weight of first complex enzyme into the extractive solution, and performing enzymolysis at pH8.5 and 54 deg.C for 2 hr. The first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease.
In the step (2), the resin adsorption time is 3 hours, and the diameter-height ratio of the used resin column is 1: 3.
In the step (2), the process conditions of the second enzymolysis are as follows: adding second complex enzyme with the weight 0.004 times of that of the pig bone, and carrying out enzymolysis treatment for 2 hours at the pH of 7.0 and the temperature of 53 ℃. The second compound enzyme comprises: 1 part of keratinase, 0.4 part of bromelain and 0.2 part of neutral protease.
In the step (2), the process conditions of the secondary shearing are as follows: the first shearing time was 30 minutes at 6000 rpm and 80 minutes at 2000 rpm.
In the step (4), the specific method of oxidation is as follows: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
In the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity (5%) and the concentration ratio (1/3 concentrated to the original volume) of the permeate reach the required range, the concentrated feed liquid is ready to be conveyed into the oxidation tank, and whether the valve switching of the oxidation tank is correct or not is checked.
In the step (4), the alcohol precipitation method specifically comprises the following steps: adding 70% ethanol water solution with volume concentration equal to the volume of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
In the step (4), the drying process conditions are as follows: dried at 55 ℃ for 8 hours.
Example 2:
a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, firstly performing first extraction at 120 ℃ for 80 minutes, and then performing second extraction at 135 ℃ for 60 minutes to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting ossein peptide with molecular weight lower than 1000Da through ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
In the step (1), crushing pig bones, adding the crushed pig bones into a first part of water with the weight 3 times that of the crushed pig bones, performing first extraction, and filtering to obtain a first extraction solution and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, carrying out secondary extraction, filtering to obtain a second extraction solution, and combining the second extraction solution with the first extraction solution to obtain the extraction solution.
The pig bone crushing method comprises the following steps: crushing fresh and clean pig bones to 5mm by using a powerful osteoclast machine, then adding the crushed pig bones into water with the weight of 1.5 times of that of the pig bones, then grinding the pig bones for 8 minutes by a wet method, centrifuging, drying and scattering the pig bones to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m. And wet grinding is carried out by adopting a colloid mill, the gap between two grinding discs of the colloid mill is 0.03mm, and the rotating speed is 12000 r/min. The process conditions of centrifugation are as follows: centrifuging at 12000r/min for 6 min, and discarding the supernatant. The drying process condition is drying at 100 ℃ for 8 hours. And scattering the dried material by using air flow.
In the step (2), before the first enzymolysis, floating oil on the surface of the extract is skimmed.
In the step (2), the process conditions of the first enzymolysis are as follows: the extract was added with 0.006 times the weight of pig bone of the first complex enzyme, and subjected to enzymatic hydrolysis at pH8.5 and 54 ℃ for 2 hours. The first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease and 0.5 part of trypsin.
In the step (2), the resin adsorption time is 5 hours, and the diameter-height ratio of the used resin column is 1: 15.
In the step (2), the process conditions of the second enzymolysis are as follows: adding second complex enzyme with the weight 0.006 times of that of pig bone, and performing enzymolysis at pH7.0 and 53 deg.C for 2 hr. The second compound enzyme comprises: keratinase 1 part, neutral protease 0.3 part.
In the step (2), the process conditions of the secondary shearing are as follows: 8000 rpm for 60 minutes, 3000 rpm for 120 minutes.
In the step (4), the specific method of oxidation is as follows: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
In the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity (6%) and the concentration ratio (1/4 concentrated to the original volume) of the permeate reach the required range, the concentrated feed liquid is ready to be delivered into the oxidation tank, and whether the valve switching of the oxidation tank is correct is checked.
In the step (4), the alcohol precipitation method specifically comprises the following steps: adding 80% ethanol water solution with volume concentration equal to the volume of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
In the step (4), the drying process conditions are as follows: drying at 60 deg.C for 12 hr.
Example 3:
a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, firstly performing first extraction for 80 minutes at 115 ℃, and then performing second extraction for 60 minutes at 130 ℃ to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting ossein peptide with molecular weight lower than 1000Da through ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
In the step (1), crushing pig bones, adding the crushed pig bones into a first part of water with the weight 2 times that of the crushed pig bones, performing first extraction, and filtering to obtain a first extraction solution and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, carrying out secondary extraction, filtering to obtain a second extraction solution, and combining the second extraction solution with the first extraction solution to obtain the extraction solution.
The pig bone crushing method comprises the following steps: crushing fresh and clean pig bones to 5mm by using a powerful osteoclast machine, then adding the crushed pig bones into water with the weight of 1 time, then carrying out wet grinding for 8 minutes, centrifuging, drying and scattering to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m. And wet grinding is carried out by adopting a colloid mill, the gap between two grinding discs of the colloid mill is 0.02mm, and the rotating speed is 12000 r/min. The process conditions of centrifugation are as follows: centrifuging at 10000r/min for 6 min, and discarding the supernatant. The drying process condition is drying for 8 hours at 90 ℃. And scattering the dried material by using air flow.
In the step (2), before the first enzymolysis, floating oil on the surface of the extract is skimmed.
In the step (2), the process conditions of the first enzymolysis are as follows: adding 0.004 times of pig bone weight of first complex enzyme into the extractive solution, and performing enzymolysis at pH8.5 and 54 deg.C for 2 hr. The first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease and 0.5 part of trypsin.
In the step (2), the resin adsorption time is 3 hours, and the diameter-height ratio of the used resin column is 1: 15.
In the step (2), the process conditions of the second enzymolysis are as follows: adding second complex enzyme with the weight 0.004 times of that of the pig bone, and carrying out enzymolysis treatment for 2 hours at the pH of 7.0 and the temperature of 53 ℃. The second compound enzyme comprises: 1 part of keratinase, 0.6 part of bromelain and 0.2 part of neutral protease.
In the step (2), the process conditions of the secondary shearing are as follows: 8000 rpm for 30 minutes and 3000 rpm for 80 minutes.
In the step (4), the specific method of oxidation is as follows: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
In the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity (6%) and concentration ratio (1/3 concentrated to original volume) of the permeate reach the required range, preparingAnd (4) conveying the concentrated feed liquid into the oxidation tank, and simultaneously checking whether the valve in the oxidation tank is switched correctly.
In the step (4), the alcohol precipitation method specifically comprises the following steps: adding 80% ethanol water solution with volume concentration equal to the volume of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
In the step (4), the drying process conditions are as follows: dried at 55 ℃ for 12 hours.
Example 4:
a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, firstly performing first extraction at 120 ℃ for 60 minutes, and then performing second extraction at 135 ℃ for 50 minutes to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting ossein peptide with molecular weight lower than 1000Da through ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
In the step (1), crushing pig bones, adding the crushed pig bones into a first part of water with the weight 3 times that of the crushed pig bones, performing first extraction, and filtering to obtain a first extraction solution and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, carrying out secondary extraction, filtering to obtain a second extraction solution, and combining the second extraction solution with the first extraction solution to obtain the extraction solution.
The pig bone crushing method comprises the following steps: crushing fresh and clean pig bones to 3mm by using a powerful osteoclast machine, then adding the crushed pig bones into water with the weight of 1.5 times of that of the pig bones, then grinding the pig bones for 5 minutes by a wet method, centrifuging, drying and scattering the pig bones to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m. And wet grinding is carried out by adopting a colloid mill, the gap between two grinding discs of the colloid mill is 0.03mm, and the rotating speed is 10000 r/min. The process conditions of centrifugation are as follows: centrifuging at 12000r/min for 5 min, and discarding the supernatant. The drying process condition is drying at 100 ℃ for 5 hours. And scattering the dried material by using air flow.
In the step (2), before the first enzymolysis, floating oil on the surface of the extract is skimmed.
In the step (2), the process conditions of the first enzymolysis are as follows: the extract was added with 0.006 times the weight of pig bone of the first complex enzyme, and subjected to enzymatic hydrolysis at pH8.5 and 54 ℃ for 2 hours. The first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease and 0.3 part of trypsin.
In the step (2), the resin adsorption time is 5 hours, and the diameter-height ratio of the used resin column is 1: 3.
In the step (2), the process conditions of the second enzymolysis are as follows: adding second complex enzyme with the weight 0.006 times of that of pig bone, and performing enzymolysis at pH7.0 and 53 deg.C for 2 hr. The second compound enzyme comprises: 1 part of keratinase, 0.4 part of bromelain and 0.3 part of neutral protease.
In the step (2), the process conditions of the secondary shearing are as follows: the first shearing time was 60 minutes at 6000 rpm and 120 minutes at 2000 rpm.
In the step (4), the specific method of oxidation is as follows: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
In the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity (5%) and the concentration ratio (1/4 concentrated to the original volume) of the permeate reach the required range, the concentrated feed liquid is ready to be conveyed into the oxidation tank, and whether the valve switching of the oxidation tank is correct or not is checked.
In the step (4), the alcohol precipitation method specifically comprises the following steps: adding 70% ethanol water solution with volume concentration equal to the volume of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
In the step (4), the drying process conditions are as follows: drying at 60 ℃ for 8 hours.
Example 5:
a method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, firstly performing primary extraction at 118 ℃ for 70 minutes, and then performing secondary extraction at 132 ℃ for 55 minutes to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting ossein peptide with molecular weight lower than 1000Da through ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
In the step (1), crushing pig bones, adding the crushed pig bones into a first part of water with the weight 2 times that of the crushed pig bones, performing first extraction, and filtering to obtain a first extraction solution and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, carrying out secondary extraction, filtering to obtain a second extraction solution, and combining the second extraction solution with the first extraction solution to obtain the extraction solution.
The pig bone crushing method comprises the following steps: crushing fresh and clean pig bones to 4mm by using a powerful osteoclast machine, then adding the crushed pig bones into water with the weight of 1.2 times of that of the pig bones, then grinding the pig bones for 6 minutes by a wet method, centrifuging, drying and scattering the pig bones to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m. And wet grinding is carried out by adopting a colloid mill, the gap between two grinding discs of the colloid mill is 0.02mm, and the rotating speed is 11000 r/min. The process conditions of centrifugation are as follows: centrifuging at 11000r/min for 6 min, and discarding the supernatant. The drying process condition is drying for 6 hours at 95 ℃. And scattering the dried material by using air flow.
In the step (2), before the first enzymolysis, floating oil on the surface of the extract is skimmed.
In the step (2), the process conditions of the first enzymolysis are as follows: adding 0.005 times of pig bone weight of first complex enzyme into the extractive solution, and performing enzymolysis at pH8.5 and 54 deg.C for 2 hr. The first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease and 0.4 part of trypsin.
In the step (2), the resin adsorption time is 4 hours, and the diameter-height ratio of the used resin column is 1: 10.
In the step (2), the process conditions of the second enzymolysis are as follows: adding second complex enzyme 0.005 times the weight of pig bone, and performing enzymolysis at pH7.0 and 53 deg.C for 2 hr. The second compound enzyme comprises: 1 part of keratinase, 0.5 part of bromelain and 0.25 part of neutral protease.
In the step (2), the process conditions of the secondary shearing are as follows: 7000 rpm for 50 minutes and 3000 rpm for 100 minutes.
In the step (4), the specific method of oxidation is as follows: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
In the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity (5%) and the concentration ratio (1/4 concentrated to the original volume) of the permeate reach the required range, the concentrated feed liquid is ready to be conveyed into the oxidation tank, and whether the valve switching of the oxidation tank is correct or not is checked.
In the step (4), the alcohol precipitation method specifically comprises the following steps: adding 74% ethanol water solution with volume concentration equal to the volume of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
In the step (4), the drying process conditions are as follows: drying at 58 ℃ for 10 hours.
Comparative example 1
A method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, firstly performing first extraction for 50 minutes at 130 ℃, and then performing second extraction for 60 minutes at 115 ℃ to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting ossein peptide with molecular weight lower than 1000Da through ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
In the step (1), crushing pig bones, adding the crushed pig bones into a first part of water with the weight 2 times that of the crushed pig bones, performing first extraction, and filtering to obtain a first extraction solution and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, carrying out secondary extraction, filtering to obtain a second extraction solution, and combining the second extraction solution with the first extraction solution to obtain the extraction solution.
The pig bone crushing method comprises the following steps: crushing fresh and clean pig bones to 3mm by using a powerful osteoclast machine, then adding the crushed pig bones into 1 time of water by weight, then grinding the pig bones for 5 minutes by a wet method, centrifuging, drying and scattering the pig bones to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m. And wet grinding is carried out by adopting a colloid mill, the gap between two grinding discs of the colloid mill is 0.02mm, and the rotating speed is 10000 r/min. The process conditions of centrifugation are as follows: centrifuging at 10000r/min for 5 min, and discarding the supernatant. The drying process condition is drying for 5 hours at 90 ℃. And scattering the dried material by using air flow.
In the step (2), before the first enzymolysis, floating oil on the surface of the extract is skimmed.
In the step (2), the process conditions of the first enzymolysis are as follows: adding 0.004 times of pig bone weight of first complex enzyme into the extractive solution, and performing enzymolysis at pH8.5 and 54 deg.C for 2 hr. The first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease and 0.3 part of trypsin.
In the step (2), the resin adsorption time is 3 hours, and the diameter-height ratio of the used resin column is 1: 3.
In the step (2), the process conditions of the second enzymolysis are as follows: adding second complex enzyme with the weight 0.004 times of that of the pig bone, and carrying out enzymolysis treatment for 2 hours at the pH of 7.0 and the temperature of 53 ℃. The second compound enzyme comprises: 1 part of keratinase, 0.4 part of bromelain and 0.2 part of neutral protease.
In the step (2), the process conditions of the secondary shearing are as follows: the first shearing time was 30 minutes at 6000 rpm and 80 minutes at 2000 rpm.
In the step (4), the specific method of oxidation is as follows: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
In the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity (5%) and the concentration ratio (1/3 concentrated to the original volume) of the permeate reach the required range, the concentrated feed liquid is ready to be conveyed into the oxidation tank, and whether the valve switching of the oxidation tank is correct or not is checked.
In the step (4), the alcohol precipitation method specifically comprises the following steps: adding 70% ethanol water solution with volume concentration equal to the volume of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
In the step (4), the drying process conditions are as follows: dried at 55 ℃ for 8 hours.
Comparative example 2
A method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones comprises the following steps:
(1) secondary extraction: adding pig bones into water, firstly performing first extraction for 60 minutes at 115 ℃, and then performing second extraction for 50 minutes at 130 ℃ to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting ossein peptide with molecular weight lower than 1000Da through ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
In the step (1), crushing pig bones, adding the crushed pig bones into a first part of water with the weight 2 times that of the crushed pig bones, performing first extraction, and filtering to obtain a first extraction solution and filter residues; and adding the filter residue into a second part of water with the same quantity as the first part of water, carrying out secondary extraction, filtering to obtain a second extraction solution, and combining the second extraction solution with the first extraction solution to obtain the extraction solution.
The pig bone crushing method comprises the following steps: crushing fresh and clean pig bones to 3mm by using a powerful osteoclast machine, then adding the crushed pig bones into 1 time of water by weight, then grinding the pig bones for 5 minutes by a wet method, centrifuging, drying and scattering the pig bones to obtain fine pig bone powder with the particle size of less than or equal to 10 mu m. And wet grinding is carried out by adopting a colloid mill, the gap between two grinding discs of the colloid mill is 0.02mm, and the rotating speed is 10000 r/min. The process conditions of centrifugation are as follows: centrifuging at 10000r/min for 5 min, and discarding the supernatant. The drying process condition is drying for 5 hours at 90 ℃. And scattering the dried material by using air flow.
In the step (2), before the first enzymolysis, floating oil on the surface of the extract is skimmed.
In the step (2), the process conditions of the first enzymolysis are as follows: adding 0.004 times of the first complex enzyme of the weight of the pig bone, and carrying out enzymolysis treatment for 2 hours at the pH of 7.0 and the temperature of 53 ℃. The second compound enzyme comprises: 1 part of keratinase, 0.4 part of bromelain and 0.2 part of neutral protease.
In the step (2), the resin adsorption time is 3 hours, and the diameter-height ratio of the used resin column is 1: 3.
In the step (2), the process conditions of the second enzymolysis are as follows: adding second complex enzyme 0.004 times of pig bone weight into the extractive solution, and performing enzymolysis at pH8.5 and 54 deg.C for 2 hr. The first complex enzyme comprises the following components in parts by weight: 1 part of alkaline protease and 0.3 part of trypsin.
In the step (2), the process conditions of the secondary shearing are as follows: the first shearing time was 30 minutes at 6000 rpm and 80 minutes at 2000 rpm.
In the step (4), the specific method of oxidation is as follows: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
In the step (4), the process conditions of the hollow fiber membrane equipment are as follows: the inlet pressure is less than or equal to 0.5MPa, the outlet pressure is less than or equal to 0.2MPa, and the feeding flow rate is less than or equal to 15m3H, permeate flow rate is more than or equal to 0.6m3H; in the operation process, feeding materials into the raw material barrel according to the mass ratio of the materials to the water of 1: 1; when the salinity (5%) and the concentration ratio (1/3 concentrated to the original volume) of the permeate reach the required range, the concentrated feed liquid is ready to be conveyed into the oxidation tank, and whether the valve switching of the oxidation tank is correct or not is checked.
In the step (4), the alcohol precipitation method specifically comprises the following steps: adding 70% ethanol water solution with volume concentration equal to the volume of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
In the step (4), the drying process conditions are as follows: dried at 55 ℃ for 8 hours.
Test examples
1. The results of examining the chondroitin sulfate yield and purity, and the collagen peptide yield and physicochemical indexes of examples 1 to 5 and comparative examples 1 to 2 are shown in Table 2.
The physicochemical index detection of the ossein peptide refers to the collagen peptide GB31645-2018 of national food safety standard, and comprises the following steps: the hydroxyproline detection method is carried out according to GB/T9695.23, the total nitrogen detection method is carried out according to GB5009.5, and the ash detection method is carried out according to GB 5009.4.
TABLE 2 examination of Process yield, purity, etc
Note: "- -" indicates the absence of this item.
The molecular weight distribution ranges of the collagen peptides obtained in examples 1 to 5 and comparative example 2 were examined and are shown in Table 3.
TABLE 3 weight average molecular weight distribution of collagen peptides
As is clear from tables 2 and 3, in examples 1 to 5, high-purity chondroitin sulfate was obtained in high yield.
Claims (10)
1. A method for simultaneously extracting chondroitin sulfate and collagen peptide from pig bones is characterized by comprising the following steps:
(1) secondary extraction: adding pig bones into water, performing first extraction for 60-80 minutes at 115-120 ℃, and then performing second extraction for 50-60 minutes at 130-135 ℃ to obtain an extraction solution;
(2) enzymolysis, adsorption and shearing: performing first enzymolysis, adsorbing chondroitin sulfate by resin, performing second enzymolysis, and performing secondary shearing to make the separated collagen peptide in a small peptide form;
(3) ceramic membrane segmentation: intercepting collagen peptides with a molecular weight of less than 1000Da (weight average) by the ceramic membrane;
(4) and (3) eluting and collecting the resin obtained in the step (2), oxidizing, desalting and concentrating by using hollow fiber membrane equipment, precipitating with ethanol, centrifuging, and drying to obtain the chondroitin sulfate.
2. The method according to claim 1, wherein in the step (1), the pig bones are firstly crushed and then added into a first part of water with the weight of 2-3 times of that of the pig bones, and the first extraction liquid and the filter residue are obtained after the first extraction and the filtration; and adding the filter residue into a second part of water with the same quantity as the first part of water, performing secondary extraction, filtering to obtain a second extract, and combining the second extract with the first extract to obtain the extract.
3. The method according to claim 1, wherein in the step (2), the process conditions of the first enzymolysis are as follows: adding a first complex enzyme with the weight of 0.004-0.006 time of the weight of the pig bones into the extract, and carrying out enzymolysis treatment for 2 hours at the pH of 8.5 and the temperature of 54 ℃.
4. The method according to claim 1, wherein the resin adsorption time is 3 to 5 hours, and the diameter-height ratio of the resin column used is 1: 3 to 15.
5. The method according to claim 1, wherein in the step (2), the process conditions of the second enzymolysis are as follows: adding a second complex enzyme with the weight 0.004-0.006 time of that of the pig bone, and carrying out enzymolysis treatment for 2 hours at the pH of 7.0 and the temperature of 53 ℃.
6. The method according to claim 1, wherein in the step (2), the process conditions of the secondary shearing are as follows: the first shearing is carried out at 6000-8000 rpm for 30-60 minutes, and the first shearing is carried out at 2000-3000 rpm for 80-120 minutes.
7. The method according to claim 1, wherein in the step (4), the specific method of oxidation is: the collected eluent is averagely divided into three parts, 2 percent, 4 percent and 6 percent of hydrogen peroxide by weight of each part are respectively added, and oxidation treatment is carried out for 4 hours under the condition of pH10.5.
8. The method according to claim 1, wherein in the step (4), the process conditions of the hollow fiber membrane device are as follows: carrying out thin-wall high-speed high; in the operation process, the mass ratio of the materials to the water is 1: 1 feeding materials into a raw material barrel; when the salinity and the concentration ratio of the permeate liquid reach the required range, the concentrated feed liquid is ready to be conveyed into the oxidation tank, and whether the switching of the valve of the oxidation tank is correct or not is checked.
9. The method as claimed in claim 1, wherein in step (4), the alcohol precipitation is carried out by the following specific method: adding 70-80% ethanol water solution with volume concentration equal to the volume concentration of the concentrated solution obtained by the hollow fiber membrane equipment, and removing the ethanol by rotary evaporation after 8 hours.
10. The method according to claim 1, wherein in the step (4), the drying process conditions are as follows: drying the mixture for 8 to 12 hours at the temperature of between 55 and 60 ℃.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114457138A (en) * | 2022-02-25 | 2022-05-10 | 浙江工业大学 | Method for removing type II collagen peptide polysaccharide |
| CN117265054A (en) * | 2023-11-21 | 2023-12-22 | 烟台融科生物科技有限公司 | Method for co-producing chondroitin sulfate and bone collagen active peptide by using chicken bones |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102293427A (en) * | 2011-07-18 | 2011-12-28 | 广东富农生物科技股份有限公司 | Cow bone broth and preparation method thereof |
| CN103397069A (en) * | 2013-08-17 | 2013-11-20 | 青岛贝尔特生物科技有限公司 | Method for producing chondroitin sulfate and realizing clean co-production of protein peptides, bone oil and bone meal |
| CN103755833A (en) * | 2014-01-03 | 2014-04-30 | 漯河双汇生物工程技术有限公司 | Method for extracting chondroitin sulfate from ossein |
| CN104418957A (en) * | 2013-08-22 | 2015-03-18 | 青岛蓝农谷农产品研究开发有限公司 | Method for comprehensively utilizing cartilage to realize clean extraction of chondroitin sulfate |
| CN104531817A (en) * | 2015-01-07 | 2015-04-22 | 吴凌天 | Combined production method of hyaluronic acid, chondroitin sulfate, collagen peptide, bone meal fodder and soap |
| CN104893314A (en) * | 2015-05-22 | 2015-09-09 | 浙江海洋学院 | Application of composite active edible film prepared from peeled shrimp processing leftovers |
| CN105104754A (en) * | 2015-09-02 | 2015-12-02 | 湖南伍星生物科技有限公司 | Bone glue enzyme hydrolyzed protein feed processed by residues obtained by extracting chondroitin sulfate and processing method for bone glue enzyme hydrolyzed protein feed |
| CN109457006A (en) * | 2018-11-26 | 2019-03-12 | 天津市天大天福生物技术有限公司 | A kind of convenient preparation method of the gradient peptide of different molecular weight mixing |
| CN109527499A (en) * | 2018-10-09 | 2019-03-29 | 临沂新程金锣肉制品集团有限公司 | A kind of processing technology that ossein voluptuousness is remarkably reinforced |
-
2019
- 2019-12-31 CN CN201911423904.4A patent/CN111533826A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102293427A (en) * | 2011-07-18 | 2011-12-28 | 广东富农生物科技股份有限公司 | Cow bone broth and preparation method thereof |
| CN103397069A (en) * | 2013-08-17 | 2013-11-20 | 青岛贝尔特生物科技有限公司 | Method for producing chondroitin sulfate and realizing clean co-production of protein peptides, bone oil and bone meal |
| CN104418957A (en) * | 2013-08-22 | 2015-03-18 | 青岛蓝农谷农产品研究开发有限公司 | Method for comprehensively utilizing cartilage to realize clean extraction of chondroitin sulfate |
| CN103755833A (en) * | 2014-01-03 | 2014-04-30 | 漯河双汇生物工程技术有限公司 | Method for extracting chondroitin sulfate from ossein |
| CN104531817A (en) * | 2015-01-07 | 2015-04-22 | 吴凌天 | Combined production method of hyaluronic acid, chondroitin sulfate, collagen peptide, bone meal fodder and soap |
| CN104893314A (en) * | 2015-05-22 | 2015-09-09 | 浙江海洋学院 | Application of composite active edible film prepared from peeled shrimp processing leftovers |
| CN105104754A (en) * | 2015-09-02 | 2015-12-02 | 湖南伍星生物科技有限公司 | Bone glue enzyme hydrolyzed protein feed processed by residues obtained by extracting chondroitin sulfate and processing method for bone glue enzyme hydrolyzed protein feed |
| CN109527499A (en) * | 2018-10-09 | 2019-03-29 | 临沂新程金锣肉制品集团有限公司 | A kind of processing technology that ossein voluptuousness is remarkably reinforced |
| CN109457006A (en) * | 2018-11-26 | 2019-03-12 | 天津市天大天福生物技术有限公司 | A kind of convenient preparation method of the gradient peptide of different molecular weight mixing |
Non-Patent Citations (2)
| Title |
|---|
| SRICHAMROEN ANCHALEE 等: "" Chondroitin sulfate extraction from broiler chicken cartilage by tissue autolysis"", 《LWT-FOOD SCIENCE AND TECHNOLOGY》 * |
| 张春林 等: ""小肽的营养功能及其制备方法"", 《中国饲料》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114457138A (en) * | 2022-02-25 | 2022-05-10 | 浙江工业大学 | Method for removing type II collagen peptide polysaccharide |
| CN117265054A (en) * | 2023-11-21 | 2023-12-22 | 烟台融科生物科技有限公司 | Method for co-producing chondroitin sulfate and bone collagen active peptide by using chicken bones |
| CN117265054B (en) * | 2023-11-21 | 2024-02-13 | 烟台融科生物科技有限公司 | Method for co-producing chondroitin sulfate and bone collagen active peptide by using chicken bones |
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