CN112121067A - Bifidobacterium infantis microcapsule and preparation method thereof - Google Patents
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Abstract
The invention belongs to the technical field of biological medicines, and particularly discloses an infant bifidobacterium microcapsule and a preparation method thereof. The microcapsule takes sodium alginate as a main wall material, the bifidobacterium infantis as a core material and trehalose as a protective agent; by adopting the microcapsule technology, the bifidobacterium infantis is encapsulated in the microcapsule, so that the microcapsule has good acid resistance and enteric solubility, the contact between the bifidobacterium infantis and the external environment is effectively isolated, and the survival rate is improved.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to an infant bifidobacterium microcapsule and a preparation method thereof.
Background
Bifidobacteria are the main beneficial flora of the human intestinal tract. The beneficial bacteria not only have unique effects on preventing and treating intestinal dysbacteriosis and other senile diseases, but also have the effects of inhibiting the growth and invasion of pathogenic bacteria, improving digestive function, providing nutrition, reducing serum cholesterol, improving body immunity, resisting tumors and the like. The bifidobacterium product is a product which is prepared by utilizing bifidobacterium which is a dominant physiological bacterium of a human body to prepare a live bacterium preparation and then returns to the original habitat (intestinal tract) of the human body through oral administration to achieve the purpose of regulating or maintaining the normal physiological function of the body, and is also called as a microecological preparation. The medical field generally considers that the micro-ecological preparation is used for regulating the functions of the digestive tract of a human body and is the most important and most promising part for developing health-care functional food. With the accumulation of studies on the health benefits of bifidobacteria and the increasing demand of modern people for healthy diet, various functional foods or medical microecological preparations using active bifidobacteria have been proposed and become popular.
The bifidobacterium strains used in the medical or health-care microecological preparation are as follows: bifidobacterium infantis, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, etc. The decrease of bifidobacterium infantis in the intestinal tracts of infants can cause the increase and the propagation of harmful bacteria and cause intestinal related diseases such as dysentery, constipation and the like. After the bifidobacterium infantis is tested by the extremely acidic condition of gastric juice of a human body, the number of the live bacteria entering the intestinal tract can be greatly reduced. The infant bifidobacterium is highly sensitive to environmental factors (heat, oxygen and the like), the live bacteria product needs to be stored at the temperature of 2-8 ℃, the requirements on storage and transportation are high, the infant bifidobacterium is not acid-resistant, the number of the live bacteria is greatly reduced after passing through gastric juice and a bile salt environment at the upper end of a digestive tract, and the curative effect of the product can be greatly influenced. Therefore, the effective embedding and addition of bifidobacterium infantis to infant formula has become a hot point of research.
Patent CN201110262354.X adopts edible gelatin gum as wall material, enzyme as cross-linking agent, and prepares nontoxic and harmless Bifidobacterium infantis microcapsule, which is suitable for this special group of infants, and is beneficial to increase the viable count of Bifidobacterium infantis in infants and promote healthy growth of infants.
Disclosure of Invention
In order to realize the aim, the invention provides an infant bifidobacterium microcapsule and a preparation method thereof, wherein the microcapsule takes sodium alginate as a main wall material, the infant bifidobacterium as a core material and trehalose as a protective agent; the method adopts microcapsule technology to encapsulate the infant bifidobacterium, ensures that the microcapsule has good acid resistance and enteric solubility, effectively isolates the infant bifidobacterium from contacting with the external environment, and improves the survival rate.
The invention adopts the following technical scheme:
in one aspect, the invention provides an infant bifidobacterium microcapsule, which comprises a core material, a protective agent and a wall material;
wherein: the core material is bifidobacterium infantis;
the protective agent is one or more of sucrose, lactose, trehalose, maltose, glucose, skimmed milk powder, soluble starch, mannitol, glycerol, sorbitol, polyethylene glycol, sodium glutamate, sodium carbonate and sodium acetate;
the wall material is a natural polymer material and comprises one or more of gelatin, Arabic gum, xanthan gum, sodium alginate, lactalbumin and chitosan.
Further, the mass ratio of the sum of the wall material and the protective agent to the core material is 1-10: 1.
Furthermore, the wall material is also coated with a coating material.
Further, the coating material is methacrylic acid-ethyl acrylate copolymer (the mass ratio of the two is 1: 1).
Further, the coating material may optionally contain a plasticizer or an anti-adhesive agent. Further, the plasticizer is one or more of triethyl citrate and dibutyl sebacate; the anti-sticking agent is one or more of talcum powder, magnesium stearate, superfine silica gel powder and monoglyceride.
Further, the bifidobacterium infantis microcapsule further comprises a curing agent. Further, the bifidobacterium infantis microcapsules optionally contain no or no detergent. Further, the curing agent is a calcium chloride solution; the detergent is a sodium chloride solution.
On the other hand, the invention provides a preparation method of the bifidobacterium infantis microcapsule, which comprises the following specific steps:
1) preparing a wall material and a protective agent into a glue solution;
2) preparing a curing agent solution;
3) preparing a washing solution;
4) preparing a coating material;
5) placing the prepared solution of the 1), the 2) and the 3) in a damp-heat sterilization cabinet for sterilization;
6) cooling, placing the cooled mixture in an environment with the temperature of 2-8 ℃, adding the bifidobacterium infantis bacterial sludge into the glue solution, and uniformly stirring and dispersing to obtain a bacterial glue solution;
7) spraying the bacterial gel solution into the curing agent solution by adopting a microcapsule granulation instrument, and slowly stirring and curing to obtain wet microcapsules;
8) washing the wet microcapsule with washing solution;
9) freeze-drying the washed wet microcapsule;
10) coating the freeze-dried microcapsule with a coating material by a fluidized bed granulator.
Further, the glue solution preparation process is as follows: adding water for injection into the wall material and the protective agent to prepare the wall material with the mass percent of 0.1-10% and the protective agent with the mass percent of 0.5-30%.
Further, the curing agent solution is prepared by the following steps: adding the curing agent into water for injection to prepare a curing agent solution with the mass percent of 0.5-10.0%.
Further, the preparation process of the washing solution is as follows: the detergent is added with water for injection to prepare the washing liquid with the mass percentage of 0.9 percent.
Further, the coating material is prepared by the following steps: and (2) adding water for injection into the methacrylic acid-ethyl acrylate copolymer aqueous dispersion with the mass ratio of 1:1 to prepare the coating material with the mass percent of 10-30%.
Further, the coating material optionally comprises 10-30% by mass of a plasticizer and 5-30% by mass of an anti-sticking agent. Further, the plasticizer is selected from one or more of triethyl citrate and dibutyl sebacate; the anti-adhesion agent is selected from one or more of talcum powder, magnesium stearate, superfine silica gel powder and monoglyceride.
Further, the solution prepared in the step 5) is placed in a moist heat sterilization cabinet for sterilization at 121 ℃ for 15 min.
Further, the bacterial strain glue solution in the step 6) has a bacterial strain glue ratio of 1: 1-10.
Further, the curing time in step 7) is 15min or more.
Further, the wet microcapsules in the step 8) are washed for 1-3 times by using a sodium chloride washing solution.
Has the advantages that:
the invention provides an infant bifidobacterium microcapsule and a preparation method thereof, wherein the microcapsule takes sodium alginate as a main wall material, the infant bifidobacterium as a core material and trehalose as a protective agent, and methacrylic acid-ethyl acrylate copolymer (1: 1) as a main coating material; by adopting a microcapsule technology, the infant bifidobacterium is encapsulated in the capsule, so that the influence of external environmental factors (heat, oxygen and the like) and the influence of gastric juice and a bile salt environment at the upper end of a digestive tract can be reduced, the requirements of normal-temperature storage and transportation are met, and the number of viable bacteria released by a product reaching the intestinal tract is increased; the embedding rate of the infant bifidobacterium microcapsules prepared by the invention can reach more than 99.0 percent, and the viable count can be kept by 40 percent after the infant bifidobacterium microcapsules are treated by gastric acid for 120 min.
The microcapsule prepared by the preparation method has uniform particle size, smooth and seamless surface, good acid resistance and enteric solubility, and fully improves the number of viable bacteria.
Drawings
FIG. 1 is an appearance diagram of a microcapsule prepared by an embodiment of the present invention;
FIG. 2 is a graph showing the change of viable cell count of the microcapsules prepared in the examples of the present invention under acidic conditions.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. These examples are intended to illustrate the invention and are not intended to limit the scope of the invention.
Example 1 preparation of Bifidobacterium infantis microcapsules
1) Preparing glue solution: weighing 9.0g of sodium alginate and 60g of trehalose, adding 600ml of water for injection, and stirring until the sodium alginate and the trehalose are completely dissolved;
2) preparing a curing liquid: weighing 15.0g of calcium chloride, adding 1000ml of water for injection, and stirring until the calcium chloride is completely dissolved;
3) preparation of a washing solution: weighing 1.8g of sodium chloride, adding 2000ml of water for injection, and stirring until the sodium chloride is completely dissolved;
4) preparing a coating material: 100ml of a 30% aqueous dispersion of a methacrylic acid-ethyl acrylate copolymer (1: 1) was taken, and 100ml of water for injection was added.
5) Sterilizing the prepared solution of the 1), 2) and 3) in a damp-heat sterilization cabinet for 15min at 121 ℃;
6) preparing a bacterial gel solution: after the sterilized glue solution is cooled, placing the glue solution in an environment with the temperature of 2-8 ℃, adding the infant bifidobacterium bacterial sludge into the glue solution according to the bacterial-to-glue ratio of 1:6, and stirring and dispersing uniformly;
7) a microcapsule granulator is adopted, a 300-micron nozzle is selected, and the frequency is set to 1200Hz, and the Electrode: 1000v, adjusting the flow rate, spraying the bacterial gel solution into the calcium chloride curing solution, slowly stirring and curing for more than 15min to obtain wet microcapsules;
8) washing the wet microcapsule with sodium chloride washing solution for 3 times;
9) freeze-drying the washed wet microcapsule;
10) and coating a layer of enteric coating material on the surface of the freeze-dried microcapsule by adopting a fluidized bed granulator, wherein the weight is increased to 10-40%.
The embedding rate of the microcapsule prepared by the method of the invention is 99.8 percent, wherein the viable count reaches 5.2 multiplied by 1010cfu/g。
Example 2
1) Preparing glue solution: weighing 9.0g of sodium alginate and 57 g of trehalose, adding 600ml of water for injection, and stirring until the sodium alginate and the trehalose are completely dissolved;
2) preparing a curing liquid: weighing 12.0g of calcium chloride, adding 1000ml of water for injection, and stirring until the calcium chloride is completely dissolved;
3) preparation of a washing solution: weighing 1.7g of sodium chloride, adding 2000ml of water for injection, and stirring until the sodium chloride is completely dissolved;
4) preparing a coating material: 100ml of a 30% aqueous dispersion of a methacrylic acid-ethyl acrylate copolymer (1: 1) was taken, and 100ml of water for injection was added.
5) Sterilizing the prepared solution of 1), 2) and 3) in a damp-heat sterilization cabinet for 15min at 120 ℃;
6) preparing a bacterial gel solution: after the sterilized glue solution is cooled, placing the glue solution in an environment with the temperature of 2-8 ℃, adding the infant bifidobacterium bacterial sludge into the glue solution according to the bacterial-to-glue ratio of 1:6, and stirring and dispersing uniformly;
7) a microcapsule granulator is adopted, a 300-micron nozzle is selected, and the frequency is set to 1200Hz, and the Electrode: 1000v, adjusting the flow rate, spraying the bacterial gel solution into the calcium chloride curing solution, slowly stirring and curing for more than 20 min to obtain wet microcapsules;
8) washing the wet microcapsule with sodium chloride washing solution for 3 times;
9) freeze-drying the washed wet microcapsule;
10) and coating a layer of enteric coating material on the surface of the freeze-dried microcapsule by adopting a fluidized bed granulator, wherein the weight is increased to 10-40%.
The embedding rate of the microcapsule prepared by the method of the invention is 99.0 percent, wherein the viable count reaches 5.7 multiplied by 1010cfu/g。
Example 3
1) Preparing glue solution: weighing 9.0g of sodium alginate, 60g of trehalose and 1.8g of water-soluble chitosan, adding 600ml of water for injection, and stirring until the sodium alginate, the trehalose and the water-soluble chitosan are completely dissolved;
2) preparing a curing liquid: weighing 15.0g of calcium chloride, adding 1000ml of water for injection, and stirring until the calcium chloride is completely dissolved;
3) preparation of a washing solution: weighing 1.8g of sodium chloride, adding 2000ml of water for injection, and stirring until the sodium chloride is completely dissolved;
4) preparing a coating material: 100ml of a 30% aqueous dispersion of a methacrylic acid-ethyl acrylate copolymer (1: 1) was taken, and 100ml of water for injection was added.
5) Sterilizing the prepared solution of the 1), 2) and 3) in a damp-heat sterilization cabinet for 15min at 121 ℃;
6) preparing a bacterial gel solution: after the sterilized glue solution is cooled, placing the glue solution in an environment with the temperature of 2-8 ℃, adding the infant bifidobacterium bacterial sludge into the glue solution according to the bacterial-to-glue ratio of 1:6, and stirring and dispersing uniformly;
7) a microcapsule granulator is adopted, a 300-micron nozzle is selected, and the frequency is set to 1200Hz, and the Electrode: 1000v, adjusting the flow rate, spraying the bacterial gel solution into the calcium chloride curing solution, slowly stirring and curing for more than 15min to obtain wet microcapsules;
8) washing the wet microcapsule with sodium chloride washing solution for 3 times;
9) freeze-drying the washed wet microcapsule;
10) and coating a layer of enteric coating material on the surface of the freeze-dried microcapsule by adopting a fluidized bed granulator, wherein the weight is increased to 10-40%.
The embedding rate of the microcapsule prepared by the method of the invention is 99.3 percent, wherein the viable count reaches 6.4 multiplied by 1010cfu/g。
Example 4
1) Preparing glue solution: weighing 9.0g of sodium alginate, 60g of soluble starch and 3.8g of xanthan gum, adding 600ml of water for injection, and stirring until the mixture is completely dissolved;
2) preparing a curing liquid: weighing 15.0g of calcium chloride, adding 1000ml of water for injection, and stirring until the calcium chloride is completely dissolved;
3) preparation of a washing solution: weighing 1.8g of sodium chloride, adding 2000ml of water for injection, and stirring until the sodium chloride is completely dissolved;
4) preparing a coating material: 100ml of 30% aqueous dispersion of methacrylic acid-ethyl acrylate copolymer (1: 1) was taken, 100ml of water for injection was added, and 20% triethyl citrate and 10% talc were added.
5) Sterilizing the prepared solution of the 1), 2) and 3) in a damp-heat sterilization cabinet for 15min at 121 ℃;
6) preparing a bacterial gel solution: after the sterilized glue solution is cooled, placing the glue solution in an environment with the temperature of 2-8 ℃, adding the infant bifidobacterium bacterial sludge into the glue solution according to the bacterial-to-glue ratio of 1:6, and stirring and dispersing uniformly;
7) a microcapsule granulator is adopted, a 300-micron nozzle is selected, and the frequency is set to 1200Hz, and the Electrode: 1000v, adjusting the flow rate, spraying the bacterial gel solution into the calcium chloride curing solution, slowly stirring and curing for more than 15min to obtain wet microcapsules;
8) washing the wet microcapsule with sodium chloride washing solution for 3 times;
9) freeze-drying the washed wet microcapsule;
10) and coating a layer of enteric coating material on the surface of the freeze-dried microcapsule by adopting a fluidized bed granulator, wherein the weight is increased to 10-40%.
The embedding rate of the microcapsule prepared by the method of the invention is 99.1 percent, wherein the viable count reaches 5.5 multiplied by 1010cfu/g。
Example 5
1) Preparing glue solution: weighing 9.0g of sodium alginate, 60g of maltose and 5.0g of whey protein, adding 600ml of water for injection, and stirring until the mixture is completely dissolved;
2) preparing a curing liquid: weighing 15.0g of calcium chloride, adding 1000ml of water for injection, and stirring until the calcium chloride is completely dissolved;
3) preparation of a washing solution: weighing 1.8g of sodium chloride, adding 2000ml of water for injection, and stirring until the sodium chloride is completely dissolved;
4) preparing a coating material: 100ml of 30% aqueous dispersion of methacrylic acid-ethyl acrylate copolymer (1: 1) was taken, 100ml of water for injection was added, and 15% dibutyl sebacate and 5% colloidal silica were added.
5) Sterilizing the prepared solution of the 1), 2) and 3) in a damp-heat sterilization cabinet for 15min at 121 ℃;
6) preparing a bacterial gel solution: after the sterilized glue solution is cooled, placing the glue solution in an environment with the temperature of 2-8 ℃, adding the infant bifidobacterium bacterial sludge into the glue solution according to the bacterial-to-glue ratio of 1:6, and stirring and dispersing uniformly;
7) a microcapsule granulator is adopted, a 300-micron nozzle is selected, and the frequency is set to 1200Hz, and the Electrode: 1000v, adjusting the flow rate, spraying the bacterial gel solution into the calcium chloride curing solution, slowly stirring and curing for more than 15min to obtain wet microcapsules;
8) washing the wet microcapsule with sodium chloride washing solution for 3 times;
9) freeze-drying the washed wet microcapsule;
10) and coating a layer of enteric coating material on the surface of the freeze-dried microcapsule by adopting a fluidized bed granulator, wherein the weight is increased to 10-40%.
The embedding rate of the microcapsule prepared by the method of the invention is 99.5 percent, wherein the viable count reaches 6.1 multiplied by 1010cfu/g。
Application examples
Placing the microcapsule particles prepared by the invention into a gastric acid solution, stirring, and detecting the viable count in the microcapsule particles respectively within 30 min, 60 min, 90 min and 120 min. The specific results are shown in FIG. 2.
Therefore, the microcapsule prepared by the invention has strong acid resistance.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. An infant bifidobacterium microcapsule is characterized by comprising a core material, a protective agent and a wall material;
wherein: the core material is bifidobacterium infantis;
the protective agent is one or more of sucrose, lactose, trehalose, maltose, glucose, skimmed milk powder, soluble starch, mannitol, glycerol, sorbitol, polyethylene glycol, sodium glutamate, sodium carbonate and sodium acetate;
the wall material is a natural polymer material and comprises one or more of gelatin, Arabic gum, xanthan gum, sodium alginate, lactalbumin and chitosan.
2. The Bifidobacterium infantis microcapsule according to claim 1, wherein the mass ratio of the sum of the wall material and the protective agent to the core material is 1-10: 1.
3. The Bifidobacterium infantis microcapsule according to claim 1, wherein the wall material is further coated with a coating material.
4. The Bifidobacterium infantis microcapsule according to claim 3, wherein the coating material is a methacrylic acid-ethyl acrylate copolymer (mass ratio of 1: 1);
the coating material may optionally contain a plasticizer or an anti-adhesive agent;
the bifidobacterium infantis microcapsule also comprises a curing agent;
the bifidobacterium infantis microcapsules optionally contain no or no detergent.
5. A method for preparing Bifidobacterium infantis microcapsules according to any of claims 1 to 4, comprising the steps of:
1) preparing a wall material and a protective agent into a glue solution;
2) preparing a curing agent solution;
3) preparing a washing solution;
4) preparing a coating material;
5) placing the prepared solution of the 1), the 2) and the 3) in a damp-heat sterilization cabinet for sterilization;
6) cooling, placing the cooled mixture in an environment with the temperature of 2-8 ℃, adding the bifidobacterium infantis bacterial sludge into the glue solution, and uniformly stirring and dispersing to obtain a bacterial glue solution;
7) spraying the bacterial gel solution into the curing agent solution by adopting a microcapsule granulation instrument, and slowly stirring and curing to obtain wet microcapsules;
8) washing the wet microcapsule with washing solution;
9) freeze-drying the washed wet microcapsule;
10) coating the freeze-dried microcapsule with a coating material by a fluidized bed granulator.
6. The method for preparing the Bifidobacterium infantis microcapsule according to claim 5, wherein the gum solution is prepared by the following steps: adding water for injection into the wall material and the protective agent to prepare the wall material with the mass percent of 0.1-10% and the protective agent with the mass percent of 0.5-30%.
7. The method for preparing Bifidobacterium infantis microcapsules according to claim 5, wherein the process for preparing said solidifying agent solution is as follows: adding a curing agent into water for injection to prepare a curing agent solution with the mass percent of 0.5-10.0%;
the preparation process of the washing solution is as follows: adding the detergent into water for injection to prepare a washing solution with the mass percent of 0.9%;
the preparation process of the coating material is as follows: taking methacrylic acid-ethyl acrylate copolymer aqueous dispersion with the mass ratio of 1:1, and adding water for injection to prepare a coating material with the mass percentage of 10-30%;
the coating material selectively comprises no plasticizer or anti-sticking agent with the mass percent of 10-30% and 5-30%.
8. The method for preparing Bifidobacterium infantis microcapsules according to claim 5, wherein the solution prepared in step 5) is sterilized in a moist heat sterilizer at 121 ℃ for 15 min.
9. The method for preparing the Bifidobacterium infantis microcapsule according to claim 5, wherein the ratio of the bacterial gel in the bacterial gel solution in step 6) is 1: 1-10.
10. The method for preparing Bifidobacterium infantis microcapsules according to claim 5, wherein the curing time in step 7) is 15min or more;
and 8) washing the wet microcapsule for 1-3 times by using a sodium chloride washing solution.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112890204A (en) * | 2021-03-15 | 2021-06-04 | 中农宠物营养研究院(江苏)有限公司 | Microcapsule using grape seed extract as prebiotics and preparation method thereof |
| CN113749258A (en) * | 2021-09-16 | 2021-12-07 | 江南大学 | Calcium alginate liquid core microcapsule and preparation method thereof |
| WO2024043845A1 (en) * | 2022-08-24 | 2024-02-29 | Bi̇oprobi̇f Sağlik Gida Arge Sanayi̇ Ve Ti̇caret Li̇mi̇ted Şi̇rketi̇ | Food supplement containing the combination of single kind and high dose probiotics in one place |
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| WO2024043845A1 (en) * | 2022-08-24 | 2024-02-29 | Bi̇oprobi̇f Sağlik Gida Arge Sanayi̇ Ve Ti̇caret Li̇mi̇ted Şi̇rketi̇ | Food supplement containing the combination of single kind and high dose probiotics in one place |
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