CN110551210A - method for extracting active ingredients from rabbit milk - Google Patents
method for extracting active ingredients from rabbit milk Download PDFInfo
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- CN110551210A CN110551210A CN201910891527.0A CN201910891527A CN110551210A CN 110551210 A CN110551210 A CN 110551210A CN 201910891527 A CN201910891527 A CN 201910891527A CN 110551210 A CN110551210 A CN 110551210A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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Abstract
The invention discloses a method for extracting active ingredients from rabbit milk, which comprises the following steps: step 1), when the rabbits produce milk, manually taking out fresh rabbit milk and placing the fresh rabbit milk in a container to prepare fresh milk for later use; step 2), filtering the fresh milk, filtering out impurities, carrying out first centrifugal operation, and selecting intermediate whey liquid; step 3), carrying out second centrifugal operation, extracting the intermediate whey liquid as in the step 2), fully stirring to form insoluble impure protein particles, and filtering out the insoluble impure protein particles; and 4), adding an alkaline solution, performing chromatography by using a gel column chromatography, concentrating by using a semi-permeable membrane method or an ultrafiltration method, and drying the concentrated solution to collect the immunoglobulin A. The method for extracting the active ingredients from the rabbit milk has simple steps, convenient operation and high extraction rate of the rabbit milk, and provides a feasible and abundant method for human immunoglobulin A utilization.
Description
Technical Field
The invention relates to the technical field of immunoglobulin extraction, in particular to a method for extracting active ingredients from rabbit milk.
Background
immunoglobulin a, abbreviation: IgA, which accounts for 10-20% of serum immunoglobulins, is present in mucosal tissues such as the digestive tract, respiratory tract and urogenital system. Mucosal tissue, which has mucosal lymphoid tissue, produces IgA to protect against pathogenic invasion, and is also found in saliva, tears and milk, especially colostrum, which has a relatively high IgA content. IgA can inhibit the adhesion of microbe to respiratory tract epithelium, slow down virus propagation, has important immune barrier effect, has antibody activity to some viruses, bacteria and common antigens, and is the first defense line for preventing pathogen from invading body. Researches show that the rabbit milk has high IgA content, and therefore, the method for extracting the active ingredients in the rabbit milk is provided.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a method for extracting active ingredients from rabbit milk.
In order to achieve the purpose, the invention adopts the following technical scheme:
A method for extracting active ingredients in rabbit milk is designed, and comprises the following steps:
Step 1), feeding juicy vegetables and fruits to the milk-producing rabbits, wherein the rabbits are bile, and in order to enable the rabbits to produce milk without influence, the milk-producing rabbits live in a quiet and dry environment, and the feeding environment temperature is 18-28 ℃; when the rabbit produces milk, the fresh rabbit milk is manually taken out and placed in a container to prepare fresh milk for later use;
Step 2), filtering fresh milk, filtering out impurities, adding a proper amount of sodium chloride solution into the milk, fully stirring, pouring into a centrifugal device, carrying out first centrifugal operation, standing and layering the centrifuged milk, placing an oil layer on the upper layer, a whey liquid in the middle, and sediment at the bottom, and selecting an intermediate whey liquid;
Step 3), pouring the prepared whey liquid into the centrifuge again, carrying out second centrifugation operation, extracting the middle whey liquid as in the step 2), adding an acid solution into the whey liquid, fully stirring to form insoluble impure protein particles, filtering the whey liquid, and filtering out the insoluble impure protein particles;
And 4) adding an alkaline solution into the solution obtained in the step 3), performing chromatography by using a gel column chromatography, concentrating by using a semi-permeable membrane method or an ultrafiltration method, drying the concentrated solution, and collecting the immunoglobulin A.
Preferably, in the step 2), the rotation speed of the centrifugation speed is 3500-.
Preferably, in the step 3), the rotation speed of the centrifugation speed is 5000-.
Preferably, in the step 3), the acidic solution is one of acetic acid and citric acid, the pH is adjusted to be 4-5.5, and the reaction temperature is 25-30 ℃.
Preferably, in the step 4), the drying method is one of a vacuum drying method, a low-temperature freeze drying method or a low-temperature freeze vacuum drying method.
The method for extracting the active ingredients from the rabbit milk has the beneficial effects that: the method for extracting the active ingredients from the rabbit milk has simple steps, convenient operation and high extraction rate of the rabbit milk, and provides a feasible and abundant method for human immunoglobulin A utilization.
Detailed Description
the invention is further illustrated by the following examples.
Example one
a method for extracting active ingredients from rabbit milk comprises the following steps:
step 1), feeding juicy vegetables and fruits to the milk-producing rabbits, wherein the rabbits are bile, and in order to enable the rabbits to produce milk without influence, the milk-producing rabbits live in a quiet and dry environment, and the feeding environment temperature is 18 ℃; when the rabbit produces milk, the fresh rabbit milk is manually taken out and placed in a container to prepare fresh milk for later use;
Step 2), filtering fresh milk, filtering out impurities, adding a proper amount of sodium chloride solution into the milk, fully stirring, pouring into a centrifugal device, carrying out first centrifugal operation, standing and layering the centrifuged milk, placing an oil layer on the upper layer, a whey liquid in the middle, and sediment at the bottom, and selecting an intermediate whey liquid;
Step 3), pouring the prepared whey liquid into the centrifuge again, carrying out second centrifugation operation, extracting the middle whey liquid as in the step 2), adding an acid solution into the whey liquid, fully stirring to form insoluble impure protein particles, filtering the whey liquid, and filtering out the insoluble impure protein particles;
and 4) adding an alkaline solution into the solution obtained in the step 3), performing chromatography by using a gel column chromatography, concentrating by using a semi-permeable membrane method or an ultrafiltration method, drying the concentrated solution, and collecting the immunoglobulin A.
Preferably, in the step 2), the rotation speed of the centrifugal speed is 3800r/min, the centrifugal time is 15min, and the centrifugal temperature is 31 ℃.
preferably, in the step 3), the rotation speed of the centrifugal speed is 5000r/min, the centrifugal time is 15min, and the centrifugal temperature is 32 ℃.
Preferably, in the step 3), the acidic solution is one of acetic acid and citric acid, the pH is adjusted to be 4, and the reaction temperature is 26 ℃.
preferably, in the step 4), the drying method is one of a vacuum drying method, a low-temperature freeze drying method or a low-temperature freeze vacuum drying method.
Example two
a method for extracting active ingredients from rabbit milk comprises the following steps:
Step 1), feeding juicy vegetables and fruits to the milk-producing rabbits, wherein the rabbits are bile, and in order to enable the rabbits to produce milk without influence, the milk-producing rabbits live in a quiet and dry environment, and the feeding environment temperature is 22 ℃; when the rabbit produces milk, the fresh rabbit milk is manually taken out and placed in a container to prepare fresh milk for later use;
step 2), filtering fresh milk, filtering out impurities, adding a proper amount of sodium chloride solution into the milk, fully stirring, pouring into a centrifugal device, carrying out first centrifugal operation, standing and layering the centrifuged milk, placing an oil layer on the upper layer, a whey liquid in the middle, and sediment at the bottom, and selecting an intermediate whey liquid;
Step 3), pouring the prepared whey liquid into the centrifuge again, carrying out second centrifugation operation, extracting the middle whey liquid as in the step 2), adding an acid solution into the whey liquid, fully stirring to form insoluble impure protein particles, filtering the whey liquid, and filtering out the insoluble impure protein particles;
And 4) adding an alkaline solution into the solution obtained in the step 3), performing chromatography by using a gel column chromatography, concentrating by using a semi-permeable membrane method or an ultrafiltration method, drying the concentrated solution, and collecting the immunoglobulin A.
Preferably, in the step 2), the rotating speed of the centrifugal speed is 4000r/min, the centrifugal time is 19min, and the centrifugal temperature is 30-45 ℃.
Preferably, in the step 3), the rotation speed of the centrifugal speed is 5500r/min, the centrifugal time is 19min, and the centrifugal temperature is 28 ℃.
Preferably, in step 3), the acidic solution is one of acetic acid and citric acid, the pH is adjusted to be 4.5, and the reaction temperature is 28 ℃.
Preferably, in the step 4), the drying method is one of a vacuum drying method, a low-temperature freeze drying method or a low-temperature freeze vacuum drying method.
EXAMPLE III
a method for extracting active ingredients from rabbit milk comprises the following steps:
step 1), feeding juicy vegetables and fruits to the milk-producing rabbits, wherein the rabbits are bile, and in order to enable the rabbits to produce milk without influence, the milk-producing rabbits live in a quiet and dry environment, and the feeding environment temperature is 23 ℃; when the rabbit produces milk, the fresh rabbit milk is manually taken out and placed in a container to prepare fresh milk for later use;
step 2), filtering fresh milk, filtering out impurities, adding a proper amount of sodium chloride solution into the milk, fully stirring, pouring into a centrifugal device, carrying out first centrifugal operation, standing and layering the centrifuged milk, placing an oil layer on the upper layer, a whey liquid in the middle, and sediment at the bottom, and selecting an intermediate whey liquid;
Step 3), pouring the prepared whey liquid into the centrifuge again, carrying out second centrifugation operation, extracting the middle whey liquid as in the step 2), adding an acid solution into the whey liquid, fully stirring to form insoluble impure protein particles, filtering the whey liquid, and filtering out the insoluble impure protein particles;
and 4) adding an alkaline solution into the solution obtained in the step 3), performing chromatography by using a gel column chromatography, concentrating by using a semi-permeable membrane method or an ultrafiltration method, drying the concentrated solution, and collecting the immunoglobulin A.
preferably, in the step 2), the rotation speed of the centrifugal speed is 4000r/min, the centrifugal time is 21min, and the centrifugal temperature is 33 ℃.
Preferably, in the step 3), the rotating speed of the centrifugal speed is 5800r/min, the centrifugal time is 21min, and the centrifugal temperature is 33 ℃.
Preferably, in step 3), the acidic solution is one of acetic acid and citric acid, the pH is adjusted to be between 5, and the reaction temperature is 27 ℃.
Preferably, in the step 4), the drying method is one of a vacuum drying method, a low-temperature freeze drying method or a low-temperature freeze vacuum drying method.
example four
a method for extracting active ingredients from rabbit milk comprises the following steps:
Step 1), feeding juicy vegetables and fruits to the milk-producing rabbits, wherein the rabbits are bile, and in order to enable the rabbits to produce milk without influence, the milk-producing rabbits live in a quiet and dry environment, and the feeding environment temperature is 27 ℃; when the rabbit produces milk, the fresh rabbit milk is manually taken out and placed in a container to prepare fresh milk for later use;
Step 2), filtering fresh milk, filtering out impurities, adding a proper amount of sodium chloride solution into the milk, fully stirring, pouring into a centrifugal device, carrying out first centrifugal operation, standing and layering the centrifuged milk, placing an oil layer on the upper layer, a whey liquid in the middle, and sediment at the bottom, and selecting an intermediate whey liquid;
Step 3), pouring the prepared whey liquid into the centrifuge again, carrying out second centrifugation operation, extracting the middle whey liquid as in the step 2), adding an acid solution into the whey liquid, fully stirring to form insoluble impure protein particles, filtering the whey liquid, and filtering out the insoluble impure protein particles;
And 4) adding an alkaline solution into the solution obtained in the step 3), performing chromatography by using a gel column chromatography, concentrating by using a semi-permeable membrane method or an ultrafiltration method, drying the concentrated solution, and collecting the immunoglobulin A.
preferably, in the step 2), the rotation speed of the centrifugal speed is 4200r/min, the centrifugal time is 23min, and the centrifugal temperature is 42 ℃.
Preferably, in the step 3), the rotating speed of the centrifugal speed is 5800r/min, the centrifugal time is 23min, and the centrifugal temperature is 42 ℃.
preferably, in step 3), the acidic solution is one of acetic acid and citric acid, the pH is adjusted to be between 5.3, and the reaction temperature is 28 ℃.
Preferably, in the step 4), the drying method is one of a vacuum drying method, a low-temperature freeze drying method or a low-temperature freeze vacuum drying method.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (5)
1. A method for extracting active ingredients from rabbit milk is characterized by comprising the following steps:
Step 1), feeding juicy vegetables and fruits to the milk-producing rabbits, wherein the rabbits are bile, and in order to enable the rabbits to produce milk without influence, the milk-producing rabbits live in a quiet and dry environment, and the feeding environment temperature is 18-28 ℃; when the rabbit produces milk, the fresh rabbit milk is manually taken out and placed in a container to prepare fresh milk for later use;
Step 2), filtering fresh milk, filtering out impurities, adding a proper amount of sodium chloride solution into the milk, fully stirring, pouring into a centrifugal device, carrying out first centrifugal operation, standing and layering the centrifuged milk, placing an oil layer on the upper layer, a whey liquid in the middle, and sediment at the bottom, and selecting an intermediate whey liquid;
step 3), pouring the prepared whey liquid into the centrifuge again, carrying out second centrifugation operation, extracting the middle whey liquid as in the step 2), adding an acid solution into the whey liquid, fully stirring to form insoluble impure protein particles, filtering the whey liquid, and filtering out the insoluble impure protein particles;
And 4) adding an alkaline solution into the solution obtained in the step 3), performing chromatography by using a gel column chromatography, concentrating by using a semi-permeable membrane method or an ultrafiltration method, drying the concentrated solution, and collecting the immunoglobulin A.
2. the method as claimed in claim 1, wherein in step 2), the rotation speed of the centrifuge is 3500-4500r/min, the centrifugation time is 15-25min, and the centrifugation temperature is 30-45 ℃.
3. the method as claimed in claim 1, wherein in step 3), the rotation speed of the centrifuge is 5000-.
4. the method for extracting active ingredients from rabbit milk according to claim 1, wherein in the step 3), the acidic solution is one of acetic acid and citric acid, the pH is adjusted to 4-5.5, and the reaction temperature is 25-30 ℃.
5. The method for extracting active ingredients from rabbit milk according to claim 1, wherein in the step 4), the drying method is one of a vacuum drying method, a low-temperature freeze-drying method or a low-temperature freeze-vacuum drying method.
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| CN201910891527.0A CN110551210A (en) | 2019-09-20 | 2019-09-20 | method for extracting active ingredients from rabbit milk |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111000974A (en) * | 2020-02-24 | 2020-04-14 | 重庆大学 | Rabbit milk active small peptide solution and extraction method and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111000974A (en) * | 2020-02-24 | 2020-04-14 | 重庆大学 | Rabbit milk active small peptide solution and extraction method and application thereof |
| CN111000974B (en) * | 2020-02-24 | 2021-07-27 | 重庆大学 | Rabbit milk active small peptide solution and extraction method and application thereof |
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