Method for extracting high-purity collagen from bovine bone

A technology of pure collagen and extraction method, which is applied in the field of high-purity collagen extraction from bovine bone, can solve the problems of wide molecular weight distribution, high protein yield, and many impurity proteins, and achieves short extraction time, improved yield and purity. , the effect of improving biological activity

Active Publication Date: 2019-02-22
GUIZHOU MOUNT FANJING ECOLOGICAL AGRI INC
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AI-Extracted Technical Summary

Problems solved by technology

Enzymatic extraction is to control a certain temperature and PH, use protease to enzymolyze raw materials, and extract collagen after impurity removal and fine filtration. The cycle is short, 1-24h e...
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Abstract

The invention relates to the technical field of bio-collagen preparation, in particular to a method for extracting high-purity collagen from bovine bone. The method includes the following steps: the step S1 of washing fresh bovine bone clean; the step S2 of putting the washed bovine bone into a reactor, adding an ethanol solution, ventilating and conducting sealed reaction to form bovine bone pulp; the step S3 of treating the bovine bone pulp with a high pressure homogenizer to form uniform bovine bone pulp; the step S4 of adding an acidic solution to the bovine bone pulp, mixing uniformly, conducting ultra-high pressure impinging stream treatment, closed reactor treatment, high-voltage pulsed electric field treatment and closed reactor treatment, conducting solid and liquid separation with a filter press to obtain an extract; the step S4 of conducting ultrafiltration treatment on the extract to collect an ultrafiltrate and conducting spray drying to obtain the collagen. The purity ofthe collagen is 98% or more, the yield of the collagen is 90% or more, the biological activity of the collagen is improved, the absorption rate of the human body is increased to more than 98.5%, and the extraction time is short.

Application Domain

Connective tissue peptidesPeptide preparation methods

Technology Topic

UltrafiltrationChemistry +14

Examples

  • Experimental program(4)
  • Comparison scheme(4)
  • Effect test(2)

Example Embodiment

[0027] Example 1
[0028] A method for extracting high-purity collagen from bovine bones includes the following steps:
[0029] S1 Wash the fresh beef bones with water to remove the meat residue on the surface, and then wash them with a citric acid solution with a mass concentration of 10%;
[0030] S2 Put the washed beef bones into the reactor and add an ethanol solution with 7 times the weight of the washed beef bones and a mass concentration of 50%. After aeration to a temperature of 130°C, stop the aeration and adjust the pressure to 95MPa for a sealed reaction for 2h Then, stir at high speed to form a slurry, and pass through a 60-mesh sieve to obtain bovine bone slurry;
[0031] S3 uses a high-pressure homogenizer to process the bovine bone slurry. The material is sprayed from the flow-limiting gap, turbulent shearing and collision, so that the material is ultra-fine into very small particles, forming a uniform bovine bone slurry;
[0032] S4 Add an organic acid solution with a mass concentration of 65% to the bovine bone slurry and mix it uniformly. After 15 minutes of ultra-high pressure impinging flow treatment, the reactor is sealed for 30 minutes, and the mixed solution is treated by a high-voltage pulse electric field for 90 seconds, and then the reactor is sealed After processing for 30 minutes, use a filter press to separate solid and liquid to obtain an extract;
[0033] In S5, the extract is processed by ultrafiltration, the ultrafiltrate is collected, and spray-dried to obtain collagen;
[0034] The ventilation refers to a mixed gas of nitrogen and carbon dioxide, wherein the ratio of the amount of nitrogen to carbon dioxide is 5:1;
[0035] The organic acid is composed of the following substances: 11kg lactic acid, 7kg tartaric acid, 2kg gallic acid, 5kg acetic acid;
[0036] The added amount of the organic acid solution is 6 times the mass of the washed bovine bone;
[0037] The pressure in the ultra-high pressure impinging stream processing is 230 MPa, and the impinging distance is 4 mm;
[0038] In the high-voltage pulsed electric field treatment, the electric field intensity is 14kV/cm, and the number of pulses is 15;
[0039] The molecular weight of the ultrafiltration membrane is 0.7 million Daltons.

Example Embodiment

[0040] Example 2
[0041] A method for extracting high-purity collagen from bovine bones includes the following steps:
[0042] S1 Wash the fresh beef bones with water to remove the meat residue on the surface, and then wash them with a citric acid solution with a mass concentration of 7%;
[0043] S2 Put the washed beef bones into the reactor and add an ethanol solution with 4 times the weight of the washed beef bones and a mass concentration of 45%. After ventilating to a temperature of 110°C, stop the aeration and adjust the pressure to 80MPa for a sealed reaction for 1h Then, stir at high speed to form a slurry, and pass through a 60-mesh sieve to obtain bovine bone slurry;
[0044] S3 uses a high-pressure homogenizer to process bovine bone slurry, and the material is sprayed from the flow-limiting gap, turbulent shearing, and collision, so that the material is ultra-fine into very small particles, forming a uniform bovine bone slurry;
[0045] S4 Add an organic acid solution with a mass concentration of 55% to the bovine bone slurry and mix it evenly. After 10 minutes of ultra-high pressure impinging stream treatment, the reactor is closed for 20 minutes, and the mixed solution is treated with a high-voltage pulsed electric field for 60 seconds, and then the reactor is closed. After processing for 20 minutes, use a filter press to separate solid and liquid to obtain an extract;
[0046] In S5, the extract is processed by ultrafiltration, the ultrafiltrate is collected, and spray-dried to obtain collagen;
[0047] The ventilation refers to a mixed gas of nitrogen and carbon dioxide, wherein the ratio of the amount of nitrogen to carbon dioxide is 3:1;
[0048] The organic acid is composed of the following substances: 8kg lactic acid, 4kg tartaric acid, 1kg gallic acid, 2kg acetic acid;
[0049] The added amount of the organic acid solution is 5 times the mass of the washed bovine bone;
[0050] The pressure in the ultra-high pressure impinging stream processing is 180 MPa, and the impinging distance is 2 mm;
[0051] In the high-voltage pulsed electric field treatment, the electric field intensity is 10kV/cm, and the number of pulses is 10;
[0052] The molecular weight of the ultrafiltration membrane is 0.1 million Daltons.

Example Embodiment

[0053] Example 3
[0054] A method for extracting high-purity collagen from bovine bones includes the following steps:
[0055] S1 Wash the fresh beef bones with water to remove the meat residue on the surface, and then wash them with a citric acid solution with a mass concentration of 8%;
[0056] S2 Put the washed beef bones into the reactor and add ethanol solution with 5 times the weight of the washed beef bones and a mass concentration of 48%. After ventilating to a temperature of 120°C, stop the aeration and adjust the pressure to 85MPa. The airtight reaction is 1.5 After h, stir at high speed to form a slurry, and pass through a 60-mesh sieve to obtain bovine bone slurry;
[0057] S3 uses a high-pressure homogenizer to process bovine bone slurry, and the material is sprayed from the flow-limiting gap, turbulent shearing, and collision, so that the material is ultra-fine into very small particles, forming a uniform bovine bone slurry;
[0058] S4 Add an organic acid solution with a mass concentration of 60% to the bovine bone slurry and mix it evenly. After 12 minutes of ultra-high pressure impinging flow treatment, the reactor is sealed for 25 minutes, and the mixed solution is treated by a high-voltage pulse electric field for 75 seconds, and then the reactor is sealed After processing for 25 minutes, use a filter press to separate solid and liquid to obtain an extract;
[0059] In S5, the extract is processed by ultrafiltration, the ultrafiltrate is collected, and spray-dried to obtain collagen;
[0060] The ventilation refers to a mixed gas of nitrogen and carbon dioxide, wherein the ratio of the amount of nitrogen to carbon dioxide is 4:1;
[0061] The organic acid is composed of the following substances: 9.5kg lactic acid, 5kg tartaric acid, 1.5kg gallic acid, 3kg acetic acid;
[0062] The added amount of the organic acid solution is 5.6 times the mass of the washed bovine bone;
[0063] The pressure in the ultra-high pressure impinging stream processing is 200 MPa, and the impinging distance is 3 mm;
[0064] In the high-voltage pulsed electric field treatment, the electric field intensity is 12kV/cm, and the number of pulses is 12;
[0065] The molecular weight of the ultrafiltration membrane is 0.5 million Daltons.

PUM

PropertyMeasurementUnit
Molecular weight0.7
Molecular weight0.1
Molecular weight0.5

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