CN111374224A - Preparation method of stable high immunoglobulin plasma protein powder - Google Patents
Preparation method of stable high immunoglobulin plasma protein powder Download PDFInfo
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- CN111374224A CN111374224A CN202010367346.0A CN202010367346A CN111374224A CN 111374224 A CN111374224 A CN 111374224A CN 202010367346 A CN202010367346 A CN 202010367346A CN 111374224 A CN111374224 A CN 111374224A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
- A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to the technical field of plasma protein powder preparation, in particular to a preparation method of stable high immunoglobulin plasma protein powder, which comprises the following steps: health quarantine, vacuum blood collection, blood anticoagulation, centrifugal separation, gradient membrane concentration, microcapsule embedding and spray drying. The plasma protein powder is prepared by embedding protein molecules in plasma protein, so that the heat stability of the immune active protein is improved, the damage of the immune globulin activity caused by high temperature in the spray drying process is avoided, the utilization rate of the protein is high, the amino acid composition is balanced, the stable immune activity is realized, and the requirements of piglets on the immune globulin and the bioactive substances can be met.
Description
Technical Field
The invention relates to the technical field of plasma protein powder preparation, in particular to a preparation method of stable high immunoglobulin plasma protein powder.
Background
The cloven-hoofed domestic animals include cattle, sheep, pigs and horses, and they are born without immunoglobulins in their serum and intestinal secretions. Because of the weak ability of newborn babies to respond to these antigens, active immunity is generated very slowly and often the animals are already ill and lethal before sufficient immune protection is generated. To compensate for this deficiency, the pups must be protected from attack by a variety of infectious pathogens by passive immunity obtained by sucking antibody-rich colostrum from birth until they develop active immunity themselves. In the actual production, under the condition that newborn piglets cannot eat colostrums, the method of feeding cow milk and milk powder is commonly used for remedying, and the method can play a certain role, but has a plurality of problems. One is that the digestibility of cow milk and sow colostrum is different, and the cow milk is directly fed, which often causes diarrhea and even death of piglets; secondly, the cow milk and the sow colostrum contain different nutrient components, and even if the piglets can survive after being fed with the cow milk, the piglets can have influence on the subsequent production performance; thirdly, the milk does not contain immune globulin (antibody), which is an important reason for death of piglets. The immune globulin is used for strengthening the milk substitute and the daily ration after weaning, so that the infection of intestinal diseases can be prevented, the incidence rate of diarrhea is reduced, and the growth speed and the production performance of newborn and weaning animals are improved.
China is the first major pig producing country in the world, and fresh blood produced by slaughtering livestock and poultry is up to over 260 million tons every year. If the abundant animal blood resources are thrown away as wastes, not only is the resources wasted greatly, but also rivers and underground water are polluted greatly. Therefore, the development and utilization of animal plasma protein powder products can solve the problem of environmental pollution, realize the development and utilization of blood resources in China, greatly improve the utilization value of byproducts such as pig blood and the like, and have great significance. At present, the development and utilization of plasma protein powder in China are only limited to simple centrifugation, filtration and drying to process the plasma protein powder into animal feed, the development and utilization level is low, the additional value is not high, and the utilization and development of animal blood in China are greatly limited. Secondly, domestic researchers directly spray-dry or evaporate-dry whole blood to prepare blood powder, but the utilization rate of protein of the prepared blood product is not high, amino acid is unbalanced, and the utilization rate is influenced. Due to the defects of the high-temperature processing technology of the plasma protein powder in the current market, the activities of immunoglobulin and other bioactive factors in the plasma protein powder are seriously damaged and even lost, and the requirements of piglets on the immunoglobulin and bioactive substances cannot be met, so that the development of the stable plasma protein powder rich in high-activity immunoglobulin is of great significance.
Disclosure of Invention
The invention aims to provide a preparation method of stable high immunoglobulin plasma protein powder, which aims to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: a preparation method of stable high immunoglobulin plasma protein powder comprises the following steps:
s1: health quarantine: quarantine is carried out on the live pigs to be slaughtered, and the live pigs qualified in quarantine can enter a slaughtering sequence;
s2: vacuum blood sampling: sampling blood of the live pigs qualified for quarantine by adopting vacuum blood sampling equipment, and placing the collected pig blood in a sealed and clean storage tank;
s3: anticoagulation of blood: adding an anticoagulant into the pig blood collected in the step S2, and storing at 0-4 ℃ for later use;
s4: centrifugal separation: separating the blood in the step S3 in a high-speed centrifuge at the rotation speed of 12000-16000rmp, centrifuging, and storing the supernatant (blood plasma) in a storage tank at 0-4 ℃;
s5: and (3) gradient membrane concentration: taking the clear liquid stored in the step S4 to be placed in a 500kDa membrane filtration system with the aperture of 750-;
s6: microcapsule embedding: adding an embedding wall material into the blood serum concentrated in the step S5, and then carrying out ultrasonic treatment for 10-30min at the ultrasonic frequency of 30-60 KHz; then homogenizing at high temperature for 1-3min under 70-120 MPa;
s7: spray drying: and (3) carrying out high-temperature spray drying on the homogenized plasma liquid in the step S6 to obtain plasma protein powder, wherein the high-temperature spray drying treatment conditions are as follows: the air inlet temperature is 170-260 ℃, the air outlet temperature is 70-100 ℃, and the viscosity is 40-1000.
According to a further scheme of the invention, in the step S6, the microcapsule embedding treatment is to add the following proportion of embedding wall materials, namely 0.1-5% of sodium alginate, 0.5-1% of β -cyclodextrin, 1-3% of glyceryl monostearate, 0.5-1% of chitosan, 0.5-1% of fructose and 1-3% of soybean oil, into each kilogram of blood.
Compared with the prior art, the preparation method of the stable high immunoglobulin plasma protein powder has the beneficial effects that a microcapsule embedding technology is adopted in the preparation process, and sodium alginate, β -cyclodextrin, glyceryl monostearate, chitosan, fructose and soybean oil are used as wall materials according to a certain proportion to embed protein molecules in plasma protein, so that the heat stability of the immune active protein of the plasma protein is improved, and the immunoglobulin activity is prevented from being damaged due to high temperature in the spray drying process.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The first embodiment is as follows:
a preparation method of stable high immunoglobulin plasma protein powder comprises the following steps:
s1: health quarantine: quarantine is carried out on the live pigs to be slaughtered, and the live pigs qualified in quarantine can enter a slaughtering sequence;
s2: vacuum blood sampling: sampling blood of the live pigs qualified for quarantine by adopting vacuum blood sampling equipment, and placing the collected pig blood in a sealed and clean storage tank;
s3: anticoagulation of blood: adding an anticoagulant into the pig blood collected in the step S2, and storing at 0-4 ℃ for later use;
s4: centrifugal separation: separating the blood in the step S3 in a high-speed centrifuge at the rotation speed of 12000-16000rmp, centrifuging, and storing the supernatant (blood plasma) in a storage tank at 0-4 ℃;
s5: and (3) gradient membrane concentration: taking the clear liquid stored in the step S4 to be placed in a 500kDa membrane filtration system with the aperture of 750-;
s6, microcapsule embedding, namely adding an embedding wall material into the concentrated blood serum in the step S5, wherein the specific microcapsule embedding treatment is to add the following embedding wall materials of 0.1-5% of sodium alginate, 0.5-1% of β -cyclodextrin, 1-3% of monostearin, 0.5-1% of chitosan, 0.5-1% of fructose and 1-3% of soybean oil into each kilogram of blood, then carrying out ultrasonic treatment for 10-30min at the ultrasonic frequency of 30-60KHz to uniformly mix the added embedding wall material auxiliary materials, and then carrying out high-temperature homogenization treatment for 1-3min at the homogenization pressure of 70-120MPa to ensure that the wall materials of the sodium alginate, β -cyclodextrin, the glyceryl monostearate, the chitosan, the fructose and the soybean oil wrap protein molecules in the blood plasma to form a stable microcapsule;
s7: spray drying: and (3) carrying out spray drying on the homogenized plasma liquid in the step S6 under the following conditions and requirements that the air inlet temperature is 170-260 ℃, the air outlet temperature is 70-100 ℃ and the viscosity is 40-1000 to obtain the required plasma protein powder.
Example two:
a preparation method of stable high immunoglobulin plasma protein powder comprises the following steps:
s1: health quarantine: quarantine is carried out on the live pigs to be slaughtered, and the live pigs qualified in quarantine can enter a slaughtering sequence;
s2: vacuum blood sampling: sampling blood of the live pigs qualified for quarantine by adopting vacuum blood sampling equipment, and placing the collected pig blood in a sealed and clean storage tank;
s3: anticoagulation of blood: adding an anticoagulant into the pig blood collected in the step S2, and storing at 0-4 ℃ for later use;
s4: centrifugal separation: separating the blood obtained in the step S3 in a high-speed centrifuge at a rotation speed of 12000rmp, centrifuging, collecting supernatant (blood plasma), storing in a storage tank at 0-4 deg.C;
s5: and (3) gradient membrane concentration: putting the clear liquid stored in the step S4 into a membrane filtration system with the pore size of 750kDa for purification and enrichment, putting the filtered liquid into a membrane filtration system with the pore size of 300kDa for membrane concentration dehydration and desalination treatment to obtain a retention solution;
s6, microcapsule embedding, namely adding an embedding wall material into the concentrated blood serum in the step S5, wherein the specific microcapsule embedding treatment is to add the following embedding wall materials of 0.1 percent of sodium alginate, 0.5 percent of β -cyclodextrin, 1 percent of glyceryl monostearate, 0.5 percent of chitosan, 0.5 percent of fructose and 1 percent of soybean oil into each kilogram of blood, then ultrasonically treating for 10min at the ultrasonic frequency of 30KHz to uniformly mix the added embedding wall material auxiliary materials, and then carrying out high-temperature homogenizing treatment for 1-3min at the homogenizing pressure of 70MPa, wherein the homogenizing aims to wrap protein molecules in the blood plasma by the wall materials of the sodium alginate, β -cyclodextrin, glyceryl monostearate, chitosan, fructose and soybean oil and the like to form a stable microcapsule;
s7: spray drying: and (3) carrying out spray drying on the homogenized plasma liquid in the step S6 under the following conditions and requirements that the air inlet temperature is 170 ℃, the air outlet temperature is 70 ℃ and the viscosity is 40 to obtain the required plasma protein powder.
Example three:
a preparation method of stable high immunoglobulin plasma protein powder comprises the following steps:
s1: health quarantine: quarantine is carried out on the live pigs to be slaughtered, and the live pigs qualified in quarantine can enter a slaughtering sequence;
s2: vacuum blood sampling: sampling blood of the live pigs qualified for quarantine by adopting vacuum blood sampling equipment, and placing the collected pig blood in a sealed and clean storage tank;
s3: anticoagulation of blood: adding an anticoagulant into the pig blood collected in the step S2, and storing at 0-4 ℃ for later use;
s4: centrifugal separation: separating the blood in the step S3 in a high-speed centrifuge at the rotation speed of 12000-16000rmp, centrifuging, and storing the supernatant (blood plasma) in a storage tank at 0-4 ℃;
s5: and (3) gradient membrane concentration: putting the clear liquid stored in the step S4 into a membrane filtration system with the pore size of 600kDa for purification and enrichment, putting the filtered liquid into a membrane filtration system with the pore size of 250kDa for membrane concentration dehydration and desalination treatment to obtain a retention solution;
s6, microencapsulation, namely adding an embedding wall material into the concentrated blood serum in the step S5, wherein the specific microencapsulation treatment is to add the following embedding wall materials of 1% of sodium alginate, 0.7% of β -cyclodextrin, 2% of glyceryl monostearate, 0.7% of chitosan, 0.7% of fructose and 2% of soybean oil into each kilogram of blood, then carrying out ultrasonic treatment for 20min at the ultrasonic frequency of 40KHz to uniformly mix the added embedding wall material and auxiliary materials, and then carrying out high-temperature homogenization treatment for 2min at the homogenization pressure of 90Mpa, wherein the homogenization aims to wrap protein molecules in the blood plasma by the wall materials of the sodium alginate, β -cyclodextrin, glyceryl monostearate, chitosan, fructose and soybean oil to form a stable microcapsule;
s7: spray drying: and (3) carrying out spray drying on the homogenized plasma liquid in the step S6 under the following conditions and requirements that the air inlet temperature is 200 ℃, the air outlet temperature is 90 ℃ and the viscosity is 500 to obtain the required plasma protein powder.
Example four:
a preparation method of stable high immunoglobulin plasma protein powder comprises the following steps:
s1: health quarantine: quarantine is carried out on the live pigs to be slaughtered, and the live pigs qualified in quarantine can enter a slaughtering sequence;
s2: vacuum blood sampling: sampling blood of the live pigs qualified for quarantine by adopting vacuum blood sampling equipment, and placing the collected pig blood in a sealed and clean storage tank;
s3: anticoagulation of blood: adding an anticoagulant into the pig blood collected in the step S2, and storing at 0-4 ℃ for later use;
s4: centrifugal separation: separating the blood in the step S3 in a high-speed centrifuge at the rotation speed of 12000-16000rmp, centrifuging, and storing the supernatant (blood plasma) in a storage tank at 0-4 ℃;
s5: and (3) gradient membrane concentration: putting the clear liquid stored in the step S4 into a membrane filtration system with the aperture of 500kDa for purification and enrichment, putting the filtered liquid into a membrane filtration system with the aperture of 150kDa for membrane concentration dehydration and desalination treatment to obtain a reserved liquid;
s6, microcapsule embedding, namely adding an embedding wall material into the concentrated blood serum in the step S5, wherein the specific microcapsule embedding treatment is to add the following embedding wall materials of 5 percent of sodium alginate, 1 percent of β -cyclodextrin, 3 percent of monostearin, 1 percent of chitosan, 1 percent of fructose and 3 percent of soybean oil into each kilogram of blood, then carry out ultrasonic treatment for 30min with the ultrasonic frequency of 60KHz to ensure that all the added embedding wall material auxiliary materials are uniformly mixed, and then carry out high-temperature homogenization treatment for 3min with the homogenization pressure of 120Mpa, wherein the aim of homogenization is to ensure that the protein molecules in the blood plasma are wrapped by the wall materials of the sodium alginate, β -cyclodextrin, the monostearin, the chitosan, the fructose, the soybean oil and the like to form a stable microcapsule;
s7: spray drying: and (3) carrying out spray drying on the homogenized plasma liquid in the step S6 under the following conditions and requirements that the air inlet temperature is 260 ℃, the air outlet temperature is 100 ℃ and the viscosity is 1000 to obtain the required plasma protein powder.
In order to further verify the nutritional value of the high immunoglobulin plasma protein powder prepared by the preparation method, the application takes the fourth example as an example, and carries out a comparative feed and blood detection test, which specifically comprises the following steps:
grouping tests: 36 healthy 21-day-old SD rats were selected and randomized into 4 groups of 3 replicates each one week after acclimation feeding, 3 replicates each. Wherein group 1 was fed basal diet as a control group; group 2 was fed 1% of the immunoglobulin rich plasma protein powder obtained in example four plus basal diet; group 3 was fed 2% of the immunoglobulin rich plasma protein powder obtained in example four plus basal diet; group 4 was fed 3% of the high immunoglobulin plasma protein powder from example four plus basal diet.
The test period is 28 days, the weight is 1 per week, the blood of the rats is extracted after the feeding period is finished and is used for blood index analysis, and indexes such as the weight of the rat end and the material weight ratio of each group are counted.
Body weight and material weight ratio data were as follows:
the blood index data are as follows:
| blood index | Unit of | Group 1 | Group 2 | Group 3 | Group 4 |
| Glutamic-pyruvic transaminase | U/L | 52.4±5.30 | 56.1±3.20 | 67.3±5.38 | 71.3±6.26 |
| Glutamic-oxalacetic transaminase | U/L | 214.7±16.47 | 234.8±19.23 | 279.2±13.57 | 299.4±13.21 |
| Glucose | mmol/L | 6.2±0.36 | 6.1±0.40 | 6.3±0.32 | 6.4±0.47 |
| Total protein | g/L | 62.07±9.20 | 64.70±9.21 | 75.0±8.24 | 79.7±9.54 |
| Albumin | g/L | 30.18±4.13 | 34.7±0.58 | 44.0±5.31 | 43.7±4.33 |
| Low density lipoprotein | mmol/L | 6.1±0.60 | 5.8±0.50 | 5.2±0.34 | 5.0±0.22 |
| High density lipoprotein | umol/L | 736.4±101.36 | 753.3±40.00 | 857.7±52.09 | 841.3±68.48 |
| ImmunoglobulinsWhite G | ug/mL | 420.6±27.44 | 503.3±22.59 | 583.4±32.81 | 630.8±30.25 |
From the results of the above table, it can be seen that:
I. during the feeding period of SD rats, the weights of the groups 2, 3 and 4 are significantly higher than that of the group 1 (control group), the feed-to-weight ratio is significantly lower than that of the group 1, the feed conversion efficiency is better than that of the group 1, and the weight increases with the increase of concentration, the feed-to-weight ratio decreases, and the feed conversion efficiency increases.
II. The SD rats of groups 2, 3 and 4 had significantly reduced low density lipoprotein and significantly increased levels of glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase, total protein, albumin and immunoglobulin G.
Thus, it was confirmed that: the high immunoglobulin plasma protein powder obtained by the invention has good thermal stability and biological activity, promotes the growth and development of organisms, and improves the related immunity of the organisms.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (5)
1. A preparation method of stable high immunoglobulin plasma protein powder is characterized by comprising the following steps:
s1: health quarantine: quarantine is carried out on the live pigs to be slaughtered, and the live pigs qualified in quarantine can enter a slaughtering sequence;
s2: vacuum blood sampling: sampling blood of the live pigs qualified for quarantine by adopting vacuum blood sampling equipment, and placing the collected pig blood in a sealed and clean storage tank;
s3: anticoagulation of blood: adding an anticoagulant into the pig blood collected in the step S2, and storing at 0-4 ℃ for later use;
s4: centrifugal separation: separating the blood obtained in the step S3 in a high-speed centrifuge, centrifuging, collecting supernatant (blood plasma) in a storage tank, and storing at 0-4 deg.C;
s5: and (3) gradient membrane concentration: taking the clear liquid stored in the step S4 to be placed in a 500kDa membrane filtration system with the aperture of 750-;
s6: microcapsule embedding: adding an embedding wall material into the blood serum concentrated in the step S5, then carrying out ultrasonic treatment, and then carrying out high-temperature homogenization treatment;
s7: spray drying: and (4) carrying out high-temperature spray drying on the homogenized plasma liquid obtained in the step S6 to obtain plasma protein powder.
2. The method as claimed in claim 1, wherein the rotation speed of the S4 step is 12000-16000 rmp.
3. The method for preparing the stable plasma protein powder with high immunoglobulin content according to claim 1, wherein the ultrasonic treatment time in the step S6 is 10-30min, the ultrasonic frequency is 30-60KHz, the high temperature homogenization treatment time is 1-3min, and the homogenization pressure is 70-120 MPa.
4. The method for preparing the stable plasma protein powder with high content of immunoglobulin according to claim 1, wherein the conditions of the high temperature spray drying in the step S7 are as follows: the air inlet temperature is 170-260 ℃, the air outlet temperature is 70-100 ℃, and the viscosity is 40-1000.
5. The method for preparing the stable plasma protein powder with high content of immunoglobulin according to claim 1, wherein the microencapsulation processing in the step of S6 is performed by adding the following ingredients, in weight, 0.1% -5% of sodium alginate, 0.5% -1% of β -cyclodextrin, 1% -3% of glyceryl monostearate, 0.5% -1% of chitosan, 0.5% -1% of fructose, and 1% -3% of soybean oil to each kilogram of blood.
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| CN102132760A (en) * | 2010-10-29 | 2011-07-27 | 成都天屹生物科技有限公司 | Method for preparing pig plasma protein powder with low ash content |
| CN102599331A (en) * | 2011-01-20 | 2012-07-25 | 天津宝迪农业科技股份有限公司 | Preparation method for high-protein low-ash animal plasma protein powder |
| CN102669411A (en) * | 2012-05-16 | 2012-09-19 | 成都连接流体分离科技有限公司 | Novel production process of high-quality plasma protein powder |
| CN103932183A (en) * | 2014-04-09 | 2014-07-23 | 吉林大学 | Forest frog oil polypeptide microencapsulation and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102132760A (en) * | 2010-10-29 | 2011-07-27 | 成都天屹生物科技有限公司 | Method for preparing pig plasma protein powder with low ash content |
| CN102599331A (en) * | 2011-01-20 | 2012-07-25 | 天津宝迪农业科技股份有限公司 | Preparation method for high-protein low-ash animal plasma protein powder |
| CN102669411A (en) * | 2012-05-16 | 2012-09-19 | 成都连接流体分离科技有限公司 | Novel production process of high-quality plasma protein powder |
| CN103932183A (en) * | 2014-04-09 | 2014-07-23 | 吉林大学 | Forest frog oil polypeptide microencapsulation and preparation method thereof |
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