CN106822912B - Production method of VB12 preparation - Google Patents
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- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7135—Compounds containing heavy metals
- A61K31/714—Cobalamins, e.g. cyanocobalamin, i.e. vitamin B12
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
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Abstract
The invention discloses a production method of a VB12 preparation, which mainly comprises the steps of VB12 preparation, VB12 and protein coordination and combination, embedding, spray drying, granulation and packaging. Aiming at the characteristic that VB12 is easy to decompose, VB12 is firstly coordinated and combined with protein, and is embedded by a protective aid and porous starch, so that VB12 can exist relatively stably, is easy to store, and can be efficiently released at a fixed point in the digestive process of the gastrointestinal tract of a human body.
Description
Technical Field
The invention belongs to the field of vitamin preparations, and particularly relates to a production method of a high-efficiency and stable VB12 preparation.
Background
VB12, also known as cobalamin, is a generic name of corrins containing metallic element cobalt, is also the only vitamin containing metallic element, and is mainly present in animal food, such as pork liver, pig heart, beef, shrimp, egg, stinky tofu, soy sauce, etc.
VB12 has vital effect on human body, and can promote the development and maturation of erythrocyte, make the hemopoietic function of organism in normal state, and prevent pernicious anemia; maintaining nervous system health; can increase the utilization rate of folic acid and promote the metabolism of carbohydrate, fat and protein; has the functions of activating amino acid, promoting nucleic acid biosynthesis, promoting protein synthesis and playing an important role in the growth and development of infants; can also metabolize fatty acid, so that fat, carbohydrate and protein are properly applied by the body; eliminate dysphoria, concentrate attention, and enhance memory and sense of balance.
The human body cannot synthesize VB12 by itself, but the daily nutritional requirement is 3-5 μ g, and thus needs to be taken from food. The VB12 form in animal food cannot be directly digested and absorbed by human body, and needs to be combined with protein. VB12 is often unstable, and its stability is mainly affected by light, heat, solution pH, reducing agents, and the like. Wherein, mecobalamin and cobamamide are sensitive to light, and aqueous solution thereof is easy to be converted into hydroxycobalamin with lower sensitivity to light at room temperature; VB12 also has the potential for decomposition during thermal processing, with different pH and temperature affecting the products of thermal decomposition, resulting in the formation of a variety of VB12 analogs; meanwhile, after VB12 enters the digestive tract of a human body, if protein combination and protection of a protection auxiliary agent are not available, decomposition and loss are easy to occur under the action of gastric acid, pepsin and trypsin.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a high-efficiency and stable production method of VB12 preparation, which adopts protein for coordination and combination, adopts a protection auxiliary agent and an embedding technology, fully improves the stability of the VB12 preparation, and simultaneously provides important technical guarantee for the fixed-point high-efficiency release of the VB12 preparation in the digestive process of human gastrointestinal tract.
In order to achieve the purpose, the invention adopts the following technical scheme:
a production method of VB12 preparation is characterized by comprising the following process steps:
(1) preparation of VB 12: inoculating propionibacterium freudenreichii to sterilized bean curd yellow water, adding 5-20% of glucose, culturing and fermenting for 24-72 h in a constant-temperature incubator at 25-35 ℃, and filtering to obtain a fermentation filtrate containing VB 12;
(2) VB12 is coordinated and combined with protein, namely α -lactalbumin solution or β -lactoglobulin solution is added into the fermentation filtrate, and the mixture is kept stand for 1 to 2 hours at room temperature in a dark condition to obtain mixed liquid of VB12 and protein, wherein α -lactalbumin or β -lactoglobulin with the concentration of 0.25 to 0.75 mu M is added into VB12 with the concentration of 1 mg/L;
(3) embedding: adding the protective additive and the porous starch into the VB12 and protein mixed solution obtained in the step (2), and uniformly mixing; wherein the protective auxiliary agent comprises skimmed milk powder, glycerol and sucrose; taking the mass of the VB12 and protein mixed liquid as a reference, the adding amount of the skimmed milk powder is 15-25%, the adding amount of the glycerol is 0.5-2%, and the adding amount of the sucrose is 6-10%; the adding amount of the porous starch is 30-50% of that of the protection auxiliary agent;
(4) spray drying: spray drying the material obtained in the step (3) by using spray drying equipment to obtain powder;
(5) and (3) granulation: adding the powder obtained in the step (4) into a granulator for granulation;
(6) and (6) packaging.
Further, in the step (1), Propionibacterium Freudenreichii (PF) is inoculated into sterilized bean curd yellow water according to the proportion of 1:300, glucose accounting for 10% of the mass of the bean curd yellow water is added, the bean curd yellow water is cultured and fermented for 48 hours in a constant temperature incubator at the temperature of 30 ℃, and residues and precipitates in the fermentation liquid are filtered out through multiple times of suction filtration, so that fermentation filtrate containing VB12 is obtained.
In the step (2), 1mg/L of VB12 is added with α -lactalbumin with the concentration of 0.25 mu M or β -lactoglobulin with the concentration of 0.25 mu M, VB12 fermentation filtrate and protein solution are mixed in a container wrapping aluminum foil paper according to the volume ratio of 1:1, and the mixture is kept stand for 1h in the dark at room temperature, so that VB12 and protein are subjected to coordination and combination to improve the stability of VB 12.
Preferably, in the step (3), a combination of VB12 and protein is adsorbed by using a protective aid and porous starch, wherein the protective aid comprises skimmed milk powder, glycerol and sucrose; on the basis of the mass of the VB12 and protein mixed solution, the adding amount of the skimmed milk powder is 20 percent, the adding amount of the glycerol is 1 percent, and the adding amount of the sucrose is 8 percent; the addition amount of the porous starch is 40 percent of the protection auxiliary agent.
And (4) controlling the air inlet temperature to be 170 ℃ and the air outlet temperature to be 65 ℃ during spray drying in the step (4).
The granulation in the step (5) comprises the following specific steps:
a. pressing down a fan and starting a cylinder;
b. opening an exhaust fan, and starting exhaust;
c. starting a fan, wherein the rotating speed of the fan is 1000 r/s;
d. heating and starting, wherein the air inlet temperature is set to 55 ℃;
e. setting the back flushing system time: and (5) carrying out back blowing for 1s and pausing for 10 s. Starting automatic back flushing;
f. starting atomization fluidization, and starting a peristaltic pump;
g. setting the material temperature, wherein the upper temperature is 40 ℃ and the lower temperature is 30 ℃; the alarm temperature is 100 ℃.
And (6) vacuum nitrogen-filled packaging.
The production method of the VB12 preparation mainly comprises the steps of preparing VB12, carrying out coordination and combination on VB12 and protein, embedding, spray drying, granulating and packaging. Aiming at the characteristic that VB12 is easy to decompose, VB12 is firstly coordinated and combined with protein, and is embedded by a protective aid and porous starch, so that VB12 can exist relatively stably, is easy to store, and can be efficiently released at a fixed point in the digestive process of the gastrointestinal tract of a human body.
α -lactalbumin is a special protein synthesized by mammary gland acinar epithelium, widely exists in mammals, and is an excellent source of essential amino acid and branched chain amino acid, β -lactoglobulin is the main whey protein component in milk, is rich in various amino acid components required by human body, and is easy to digest and absorb, so α -lactalbumin and β -lactoglobulin are the most suitable proteins for combining with VB 12.
The VB12 preparation is characterized in that protein is coordinated and combined according to a special structure of VB12, a protection auxiliary agent and an embedding technology are adopted, the stability of the VB 3526 preparation is fully improved, and the VB12 preparation can be efficiently released at a fixed point in the gastrointestinal digestion process of a human body. A large number of irradiation or heat treatment experiments prove that the decomposition rate of the protein-bound VB12 is obviously lower than that of an unprotected control group under the influence of light irradiation or heat treatment, which shows that the protein-bound VB12 can effectively improve the stability of VB 12.
Compared with the prior art, the invention has the following advantages:
(1) the invention adopts the mode of coordination and combination of VB12 and protein, reduces the loss of VB12 under the influence of external factors to the greatest extent, and improves the stability of the protein.
(2) The invention applies the embedding technology to the combination of VB12 and protein, has obvious protection effect in the processes of spray drying, granulation and storage, and can achieve the effect of slow release.
Detailed Description
The present invention is further illustrated by the following examples.
Example 1
1) Preparation of VB12
Inoculating Propionibacterium Freudenreichii (PF) into sterilized bean curd yellow water at a ratio of 1:300, adding glucose 10% of the weight of the bean curd yellow water, culturing and fermenting in a 30 deg.C incubator for 48h, and filtering to remove residue and precipitate in the fermentation liquid by multiple suction filtration to obtain fermentation filtrate containing VB 12.
2) VB12 is coordinately bound with protein
Adding VB12 fermentation filtrate and α -lactalbumin into a beaker coated with aluminum foil paper, adopting 0.25 mu M concentration of α -lactalbumin for 1mg/L of VB12, mixing in the beaker coated with the aluminum foil paper according to the volume ratio of 1:1, and standing at room temperature in the dark for 1 h.
3) Embedding
Adding a protection aid and porous starch into the fermentation filtrate in proportion, wherein the protection aid comprises the following components by mass based on the mixed liquid of VB12 and protein: the additive comprises 20% of skimmed milk powder, 1% of glycerol, 8% of sucrose and 40% of porous starch.
4) Spray drying
Spray drying the fermentation filtrate obtained in the step 3) by using spray drying equipment to obtain powder, and controlling the air inlet temperature to be 170 ℃ and the air outlet temperature to be 65 ℃.
5) Granulating
Adding the powder obtained in the step 4) into a granulator for granulation, and specifically comprising the following steps:
a. and (5) pressing a fan downwards and starting the air cylinder.
b. And opening the exhaust fan, and starting exhaust.
c. And starting the fan, wherein the rotating speed of the fan is 1000 r/s.
d. Heating is started, and the air inlet temperature is set to be 55 ℃.
e. Setting the back flushing system time: and (5) carrying out back blowing for 1s and pausing for 10 s. And starting automatic back flushing.
f. Starting atomization fluidization and starting a peristaltic pump.
g. The material temperature was set at 40 ℃ for the upper temperature and 30 ℃ for the lower temperature. The alarm temperature is 100 ℃.
6) Package (I)
And (5) carrying out vacuum nitrogen-filled packaging on the product obtained in the step 5).
Example 2
1) Preparation of VB12
Inoculating Propionibacterium Freudenreichii (PF) into sterilized bean curd yellow water at a ratio of 1:300, adding glucose 10% of the weight of the bean curd yellow water, culturing and fermenting in a 30 deg.C incubator for 48h, and filtering to remove residue and precipitate in the fermentation liquid by multiple suction filtration to obtain fermentation filtrate containing VB 12.
2) VB12 is coordinately bound with protein
Adding VB12 fermentation filtrate and β -lactoglobulin into a beaker coated with aluminum foil paper, adopting 0.25 mu M concentration of β -lactoglobulin for 1mg/L VB12, mixing in the beaker coated with the aluminum foil paper according to the volume ratio of 1:1, and standing at room temperature in the dark for 1 h.
3) Embedding
Adding a protection aid and porous starch into the fermentation filtrate in proportion, wherein the protection aid comprises the following components by mass based on the mixed liquid of VB12 and protein: the additive comprises 20% of skimmed milk powder, 1% of glycerol, 8% of sucrose and 40% of porous starch.
4) Spray drying
Spray drying the fermentation filtrate obtained in the step 3) by using spray drying equipment to obtain powder, and controlling the air inlet temperature to be 170 ℃ and the air outlet temperature to be 65 ℃.
5) Granulating
Adding the powder obtained in the step 4) into a granulator for granulation, and specifically comprising the following steps:
a. and (5) pressing a fan downwards and starting the air cylinder.
b. And opening the exhaust fan, and starting exhaust.
c. And starting the fan, wherein the rotating speed of the fan is 1000 r/s.
d. Heating is started, and the air inlet temperature is set to be 55 ℃.
e. Setting the back flushing system time: and (5) carrying out back blowing for 1s and pausing for 10 s. And starting automatic back flushing.
f. Starting atomization fluidization and starting a peristaltic pump.
g. The material temperature was set at 40 ℃ for the upper temperature and 30 ℃ for the lower temperature. The alarm temperature is 100 ℃.
6) Package (I)
And (5) carrying out vacuum nitrogen-filled packaging on the product obtained in the step 5).
Claims (7)
1. A production method of VB12 preparation is characterized by comprising the following process steps:
(1) preparation of VB 12: inoculating propionibacterium freudenreichii to sterilized bean curd yellow water, adding 5-20% of glucose, culturing and fermenting for 24-72 h in a constant-temperature incubator at 25-35 ℃, and filtering to obtain a fermentation filtrate containing VB 12;
(2) VB12 is coordinated and combined with protein, namely α -lactalbumin solution or β -lactoglobulin solution is added into the fermentation filtrate, and the mixture is kept stand for 1 to 2 hours at room temperature in a dark condition to obtain mixed liquid of VB12 and protein, wherein α -lactalbumin or β -lactoglobulin with the concentration of 0.25 to 0.75 mu M is added into VB12 with the concentration of 1 mg/L;
(3) embedding: adding the protective additive and the porous starch into the VB12 and protein mixed solution obtained in the step (2), and uniformly mixing; wherein the protective auxiliary agent comprises skimmed milk powder, glycerol and sucrose; taking the mass of the VB12 and protein mixed liquid as a reference, the adding amount of the skimmed milk powder is 15-25%, the adding amount of the glycerol is 0.5-2%, and the adding amount of the sucrose is 6-10%; the adding amount of the porous starch is 30-50% of that of the protection auxiliary agent;
(4) spray drying: spray drying the material obtained in the step (3) by using spray drying equipment to obtain powder;
(5) and (3) granulation: adding the powder obtained in the step (4) into a granulator for granulation;
(6) and (6) packaging.
2. A production method of VB12 preparation according to claim 1, wherein in the step (1), the propionibacterium freudenreichii is inoculated at a ratio of 1:300, the glucose addition ratio is 10% of the mass of the soybean curd yellow water, and the mixture is cultured and fermented in a 30 ℃ constant temperature incubator for 48 hours.
3. The method for producing a VB12 preparation according to claim 1, wherein in the step (2), the volume ratio of the fermentation filtrate to the α -lactalbumin solution or β -lactoglobulin solution is 1: 1.
4. A production method of VB12 preparation according to claim 1, wherein in the step (3), the addition amount of skimmed milk powder is 20%, the addition amount of glycerol is 1% and the addition amount of sucrose is 8% based on the mass of the VB12 and protein mixed solution; the addition amount of the porous starch is 40 percent of the protection auxiliary agent.
5. The method for producing a VB12 preparation according to claim 1, wherein the inlet air temperature is controlled to be 170 ℃ and the outlet air temperature is controlled to be 65 ℃ during the spray drying in the step (4).
6. The method for producing a VB12 formulation according to claim 1, wherein the granulation in step (5) comprises the following steps:
a. pressing down a fan and starting a cylinder;
b. opening an exhaust fan, and starting exhaust;
c. starting a fan, wherein the rotating speed of the fan is 1000 r/s;
d. heating and starting, wherein the air inlet temperature is set to 55 ℃;
e. setting the back flushing system time: back blowing for 1s and pausing for 10 s; starting automatic back flushing;
f. starting atomization fluidization, and starting a peristaltic pump;
g. setting the material temperature, wherein the upper temperature is 40 ℃ and the lower temperature is 30 ℃; the alarm temperature is 100 ℃.
7. A method of producing a VB12 formulation according to claim 1, wherein step (6) is vacuum nitrogen-filled packaging.
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| CN108975964A (en) * | 2018-05-22 | 2018-12-11 | 浙江工商大学 | The preparation method of vitamin B12 Liquid organic fertilizer and the organic fertilizer culture are rich in the application of vitamin B12 vegetables |
| CN109453171A (en) * | 2018-11-13 | 2019-03-12 | 浙江工商大学 | A kind of protective agent and the preparation method and application thereof for inhibiting PM2.5 to induce intestinal inflammatory |
| CN109825543B (en) * | 2018-11-24 | 2021-04-30 | 浙江华康药业股份有限公司 | Method for fermenting vitamin B12 by xylose mother liquor microorganism based on illumination regulation |
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Non-Patent Citations (3)
| Title |
|---|
| "Effects of pH and Electrodialysis on the Binding of Vitamin B12 by β-Lactoglobulin and Associated Peptides";JAMES R. KIRK et al.;《The Journal of Nutrition》;19720601;第102卷(第6期);第699-705页 * |
| "Enhancing the vitamin B12 production and growth of Propionibacterium freudenreichii in tofu wastewater via a light-induced vitamin B12 riboswitch";Yue Yu et al.;《Appl Microbiol Biotechnol》;20150915;第99卷;第10481-10488页 * |
| "钙诱导乳清分离蛋白包埋体系对VB12的稳态研究";柴智等;《食品科学》;20101231;第31卷(第3期);第1-4页 * |
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