CN113694086A - Probiotics capsule and preparation method thereof - Google Patents
Probiotics capsule and preparation method thereof Download PDFInfo
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- CN113694086A CN113694086A CN202010431387.1A CN202010431387A CN113694086A CN 113694086 A CN113694086 A CN 113694086A CN 202010431387 A CN202010431387 A CN 202010431387A CN 113694086 A CN113694086 A CN 113694086A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4816—Wall or shell material
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4816—Wall or shell material
- A61K9/4825—Proteins, e.g. gelatin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention provides a probiotic capsule, which comprises an inner core and an outer shell coating the inner core; wherein, the material of the inner core comprises probiotics, and the material of the outer shell comprises a probiotic protective agent and zein. The structure of the probiotic capsule forms a core-shell structure of 'core-shell', and the probiotic is taken as a core material, so that the probiotic is protected by a shell material in the capsule preparation and acting on the large intestine, and the tolerance of the probiotic is improved; the shell material comprises a probiotic protective agent and zein, so that the performance of probiotics is further protected, the probiotics can be kept in good stability in the preparation process of the probiotic capsule, the tolerance of the probiotics in the intestines and stomach during working is improved, the probiotics capsule is prevented from being inactivated due to the acidic environment of the stomach, the probiotics capsule is released in the large intestine, and the effect of the probiotics is improved.
Description
Technical Field
The invention relates to the field of probiotics, in particular to a probiotic capsule and a preparation method thereof.
Background
The probiotics are active microorganisms which play a role by improving the balance of host intestinal microbial flora, have the functions of regulating the immune function of host mucous membrane and system or promoting nutrient absorption and keeping the intestinal health by regulating the balance of the microbial flora in the intestinal tract, have the safety effects of improving the gastrointestinal tolerance of organisms, clearing or reducing the adhesion of pathogenic bacteria and inhibiting the pathogenic bacteria, and are widely applied to various fields of food, medical health care, feed and the like. However, probiotics are extremely sensitive to the adverse environment of the outside world, and the survival rate of the probiotics is affected by a plurality of factors from processing to eating. Meanwhile, in the process of taking the probiotics by organisms, the acidic environment of gastric juice can also influence the survival rate of the probiotics. Most probiotics do not successfully colonize the colon and regulate the intestinal environment temporarily as they are inactivated by the inability to withstand the acidic environment of the stomach.
At present, common means for improving the tolerance of probiotics comprise pre-culture, screening resistant strains, adding protective agents and embedding. The embedding is the most common and effective method, the most common embedding technology used at present is the embedding by adopting sodium alginate, the embedding treatment aims at forming a compact physical barrier around probiotic cells, and harmful substances such as air, gastric acid, bile salt and the like can be effectively resisted or delayed to permeate, so that the damage of adverse environments to the cells is reduced. Sodium alginate is the most commonly used probiotic bacteria embedding wall material, has the advantages of low price, applicability and high biocompatibility, can form ionic colloid, is stable in an acid environment and can be dissolved in a neutral or high-pH environment, but the porous gel network structure of the sodium alginate colloid can not completely and effectively limit small-molecule harmful substances, and the action effect of the sodium alginate is very limited.
Disclosure of Invention
The invention aims to provide a probiotic capsule and a preparation method thereof, and aims to solve the problems of poor tolerance and low survival rate of probiotics in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, there is provided a probiotic capsule comprising an inner core and an outer shell surrounding the inner core; wherein, the material of the inner core comprises probiotics, and the material of the outer shell comprises a probiotic protective agent and zein.
In a second aspect, a method for preparing a probiotic capsule is provided, which comprises the following steps:
mixing a probiotic solution and a probiotic protective agent to obtain a first mixed solution, adjusting the pH of the first mixture to 3.8-4.5, standing, performing centrifugal treatment, collecting a first mixture precipitate, and performing heavy suspension treatment on the first mixture precipitate to obtain a second mixture;
providing a zein solution, adding the second mixture into the zein solution within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain the probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of probiotic bacteria protective agent and zein.
The probiotic capsule comprises an inner core and an outer shell coating the inner core, so that a core-shell structure is formed, and further, the probiotic is used as an inner core material, so that the probiotic is protected by the outer shell material in the capsule preparation and acting on the large intestine, and the tolerance of the probiotic is improved; the shell material comprises a probiotic protective agent and a zein solution, the probiotic protective agent is used as the shell material of the probiotic capsule, the performance of probiotics is further protected, the probiotics keeps good stability in the preparation process of the probiotic capsule, and meanwhile, the tolerance of the probiotics during working in intestines and stomach is guaranteed to be improved, and the probiotics cannot be inactivated due to the acidic environment of the stomach; secondly, zein is used as a shell material of the probiotic capsule, is a hydrophobic macromolecule, has good biocompatibility and degradability, can be used as a carrier of probiotics to embed the probiotics, is rich in sulfur amino acid, and is connected with protein molecules through stronger disulfide bonds and hydrophobic bonds, so that the shell structure of the prepared probiotic capsule is smooth and flat, has low oxygen permeability, can effectively prevent the contact of the medicine and external oxygen, improves the storage time of the core material probiotics, simultaneously ensures that the tolerance is improved when the capsule works in intestines and stomach, cannot be inactivated due to the acidic environment of the stomach, ensures that the probiotic capsule is released in the large intestine, and improves the effect of the probiotics.
The preparation method of the probiotic capsule comprises the steps of providing a probiotic solution, mixing the probiotic solution with a probiotic protective agent to obtain a first mixture, attaching the first mixture to the surface of probiotic through a positive and negative charge combination reaction to form a protective layer, adding a zein solution, combining the zein solution with the probiotic protective agent to form a shell material of the probiotic capsule, and protecting a core material probiotic solution; the preparation method has simple process and mild conditions in the preparation process, and is beneficial to wide application.
Drawings
Fig. 1 is a schematic structural diagram of a probiotic capsule provided in example 1 of the present invention.
Fig. 2 shows the content of beneficial bacteria in the in vitro simulated release process of the probiotic capsule provided in examples 1 to 4 of the present invention.
Detailed Description
In order to make the objects, technical solutions and technical effects of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments obtained by a person of ordinary skill in the art without any inventive work in conjunction with the embodiments of the present invention are within the scope of the embodiments of the present invention.
In the description of the embodiments of the present invention, it should be understood that the terms "first", "second", and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the embodiments of the present invention, "a plurality" means two or more unless specifically limited otherwise.
In a first aspect, an embodiment of the present invention provides a probiotic capsule, as shown in fig. 1, the probiotic capsule includes an inner core and an outer shell covering the inner core; wherein, the inner core comprises probiotics, and the outer shell comprises probiotics protective agent and zein.
The probiotic capsule comprises an inner core and an outer shell coating the inner core, so that a core-shell structure is formed, and further, the probiotic is used as an inner core material, so that the probiotic is protected by the outer shell material in the capsule preparation and acting on the large intestine, and the tolerance of the probiotic is improved; the shell material comprises a probiotic protective agent and a zein solution, the probiotic protective agent is used as the shell material of the probiotic capsule, the performance of the probiotics is further protected, the probiotics keeps good stability in the preparation process of the probiotic capsule, and meanwhile, the tolerance of the probiotics during the operation in intestines and stomach is ensured to be improved, and the probiotics cannot be inactivated due to the acidic environment of the stomach; secondly, zein is used as a shell material of the probiotic capsule, the zein is a hydrophobic macromolecule, has good biocompatibility and degradability, can be used as a carrier of probiotics to embed the probiotics, and is rich in sulfur amino acid, and protein molecules are connected by strong disulfide bonds and hydrophobic bonds, so that the shell structure of the prepared probiotic capsule is smooth and flat, and meanwhile, the zein has low permeability to oxygen, can effectively prevent the medicine from contacting with external oxygen, improves the storage time of the core material probiotics, and simultaneously ensures that the tolerance is improved when the capsule works in intestines and stomach, and the capsule cannot be inactivated due to the acidic environment of the stomach, thereby ensuring the release of the probiotic capsule in the large intestine and improving the effect of the probiotics.
Specifically, the core material of the probiotic capsule comprises probiotics, and the probiotics need to be subjected to certain oxygen-resistant acclimation culture, so that the obtained probiotics have a high survival rate in the aerobic operation process. Preferably, the probiotic bacteria are selected from any one of lactic acid bacteria, lactobacillus, rhamnosus, etc.
Specifically, in the probiotic capsule, the shell material comprises a probiotic protectant and zein. The probiotic protective agent is used as a shell material to further protect the performance of probiotics, and secondly, zein is used as the shell material of the probiotic capsule and is synergistically acted with the probiotic protective agent to prepare the protective shell of the probiotic capsule, so that the probiotics can keep good stability in the preparation process of the probiotic capsule, and meanwhile, the tolerance of the probiotics in the gastrointestinal tract is guaranteed to be improved, and the probiotics cannot be inactivated due to the acidic environment of the stomach.
Preferably, the probiotic protecting agent comprises the following components in parts by weight: 5-50 parts of skimmed milk powder or whey powder; 0.5-10 parts of trehalose; 0.5-10 parts of glycerol; 0.5-10 parts of sodium glutamate; 1-15 parts of maltodextrin. The probiotic protective agent comprises 5-50 parts of skimmed milk powder or whey powder, the protein content of the skimmed milk powder or whey powder is 25% -30%, the protein content with higher concentration can provide corresponding protection for the probiotics, and the probiotics cannot be influenced by external force or external conditions to reduce the survival rate of the probiotics in the subsequent preparation process of the capsule, such as freeze drying. Furthermore, the probiotic protective agent comprises 0.5-10 parts of trehalose, the trehalose is non-reducing sugar formed by two glucose molecules through 1, 1-glycosidic bonds, and the trehalose can form a unique protective film on the surface of active substances such as probiotics and the like, so that the probiotic molecules are effectively protected from inactivation. Furthermore, the probiotic protective agent comprises 0.5-10 parts of glycerol, and the addition of the glycerol can improve the tolerance activity of the probiotics, ensure that the probiotics keep activity under the freeze-drying treatment condition, and avoid low-temperature influence. Furthermore, the probiotic protective agent also comprises 0.5-10 parts of sodium glutamate and 1-15 parts of maltodextrin, and the sodium glutamate and the maltodextrin are added, so that the tolerance of the probiotics can be guaranteed, and the survival rate of the probiotics is improved.
Specifically, the material of the shell also comprises zein. Zein is a hydrophobic macromolecule, has good biocompatibility and degradability, can be used as a carrier of probiotics, and forms embedding for the probiotics, because zein is rich in sulfur-containing amino acid, protein molecules are connected by stronger disulfide bonds and hydrophobic bonds, so that the shell structure of the prepared probiotic capsule is smooth and flat, and meanwhile, the zein has low oxygen permeability, can effectively prevent the medicine from contacting with external oxygen, improves the storage time of the core material probiotics, and simultaneously ensures that the tolerance is improved when the zein works in intestines and stomach, and the zein cannot be inactivated due to the acid environment of stomach, ensures that the probiotic capsule is released in the large intestine, and improves the effect of the probiotics.
Preferably, the mass ratio of the shell to the core is (2-3): 1. the mass ratio of the inner core to the outer shell of the probiotic capsule is controlled, so that the inner core and the outer shell can form better interaction, the survival rate of the probiotics is further ensured to be high, the prepared probiotic capsule is also ensured not to be inactivated due to the acidic environment of the stomach, the probiotic capsule is ensured to be released in the large intestine, and the action effect of the probiotics is improved. If the addition amount of the shell is too high, the net content of probiotics in the prepared probiotic capsule is too low, and the action effect of the probiotic capsule is influenced; if the addition amount of the shell is too low, the embedding efficiency of the prepared probiotic capsule is poor, and the survival rate of the probiotics is affected. Further preferably, the mass ratio of the shell to the core is 2: the probiotic capsule has the advantages that 1, the addition amount of the inner core and the outer shell is controlled, the inner core and the outer shell can form better interaction, the embedding effect is better, the tolerance of the probiotic capsule is improved when the probiotic capsule works in intestines and stomach, the probiotic capsule cannot be inactivated due to the acidic environment of the stomach, the probiotic capsule is released in the large intestine, and the effect of probiotics is improved.
Preferably, the particle size of the inner core is 1.5-1.8 μm, and the thickness of the outer shell is 0.5-0.8 μm. The particle size of the inner core is controlled to be 1.5-1.8 mu m, the thickness of the shell can be controlled, the particle size of the prepared probiotic capsule is moderate, and the optimal effect can be realized. If the particle size is too large, the digestion of the probiotic capsule in the digestive tract in vivo is affected, and if the particle size is too small, the effect of the probiotic is affected, and the release of the probiotic in the large intestine is affected. In a specific embodiment of the present invention, the particle size of the inner core is controlled to be 1.5 μm.
Furthermore, the thickness of the shell is controlled to be 0.5-0.8 μm. The prepared probiotic capsule is ensured to have moderate particle size, and the best effect can be realized. In a specific embodiment of the invention, the thickness of the shell is 0.5 μm.
The probiotic capsule is prepared by the following preparation method of the probiotic capsule.
In a second aspect, the embodiment of the present invention further provides a preparation method of a probiotic capsule, including the following steps:
s01, mixing the probiotic solution and the probiotic protective agent to obtain a first mixed solution, adjusting the pH value of the first mixture to 3.8-4.5, standing, performing centrifugal treatment, collecting the first mixture precipitate, and performing heavy suspension treatment on the first mixture precipitate to obtain a second mixture;
s02, providing a zein solution, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture;
s03, freeze-drying the third mixture to obtain a probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein.
According to the preparation method of the probiotic capsule, provided is a probiotic solution, the probiotic solution is mixed with a probiotic protective agent to obtain a first mixture, the first mixture is attached to the surface of probiotic through a positive and negative charge combination reaction to form a protective layer, then a zein solution is added, the zein solution is combined with the probiotic protective agent to form a shell material of the probiotic capsule, and the core material, namely probiotic, is protected; the preparation method has simple process and mild conditions in the preparation process, and is beneficial to wide application.
Specifically, in step S01, a probiotic solution is provided, and the preparation method of the probiotic solution includes performing polar fermentation culture on the probiotic with a liquid culture medium, performing centrifugal concentration, and resuspending in sterile water to obtain the probiotic solution.
Further, a probiotic protective agent is provided, and preferably, the probiotic protective agent comprises the following components in parts by weight: 5-50 parts of skimmed milk powder or whey powder; 0.5-10 parts of trehalose; 0.5-10 parts of glycerol; 0.5-10 parts of sodium glutamate; 1-15 parts of maltodextrin. In a preferred embodiment of the invention, the probiotic protective agent comprises 5-50 parts of skimmed milk powder and 0.5-10 parts of trehalose; 0.5-10 parts of glycerol.
Further, adding the probiotic protective agents into the probiotic solution one by one, mixing to obtain a first mixture, preferably, performing vortex oscillation on the first mixture for 10 minutes to ensure that the probiotic protective agents can be fully dissolved in the probiotic solution.
Preferably, in the step of adjusting the pH of the first mixture to 3.8 to 4.5, the first mixed solution is dripped into a buffer solution, and the pH of the first mixture is adjusted to 3.8 to 4.5; wherein the buffer solution is at least one selected from potassium hydrogen phthalate solution, disodium hydrogen phosphate-citric acid buffer solution and citric acid-sodium citrate buffer solution. And controlling the pH value of the first mixture to be 3.8-4.5 to ensure that the survival rate of the probiotics is higher under the buffering of the pH condition and the activity of the probiotics is not influenced.
Preferably, the concentration of the probiotics in the first mixture is 1-9 x 1010And CFU/mL, wherein the concentration of the probiotics in the first mixture is controlled to be higher, so that certain loss of the probiotics in the processing process is prevented, and the concentration of the probiotics in the finally obtained second mixture is higher. Further preferably, the mass ratio of the probiotics to the buffer solution in the first mixed solution is 1: (2-2.5). The adding proportion of the buffer solution and the probiotics in the first mixed solution is controlled, so that the pH value of a solution system can be well controlled, and meanwhile, the mixed solution does not contain excessive buffer solution and excessive impurities.
More preferably, in the step of adding the first mixed solution dropwise to the buffer solution, the dropwise adding speed is 1 to 1.5 drops/second. The addition control mode is a dropwise addition mode, the contact area is controlled to be larger, and the reaction is more complete.
Further, after the pH value of the first mixture is adjusted to 3.8-4.5, standing treatment is carried out. Preferably, the standing treatment time is 30-40 minutes, and the purpose of the standing treatment is to fully mix the probiotic solution and the probiotic protective agent, and attach the mixture to the surface of the probiotic through a positive and negative charge combination reaction to form a protective layer, so that the probiotic is protected from being influenced by subsequent operation.
Further, the first mixture precipitate is collected by centrifugation after the standing treatment, and preferably, the centrifugation conditions are as follows: and the centrifugal speed is 1000-1200 rpm, the centrifugal temperature is 4 ℃, the centrifugal time is 30-35 minutes, and the centrifugal treatment is carried out under the conditions, so that the probiotic precipitation can be ensured, meanwhile, redundant probiotic protective agents and buffer solutions are removed, and the obtained first mixture precipitation is ensured not to contain other impurities. The conditions of the centrifugal treatment are controlled as above, so that the conditions of the centrifugal treatment are mild, and the activity of the probiotics is ensured.
Further preferably, the obtained first mixture precipitate is collected, and the first mixture precipitate is washed with sterile physiological saline to ensure that the first mixture precipitate is free from other impurities.
Further, carrying out heavy suspension treatment on the first mixture precipitate to obtain a second mixture; controlling the concentration of beneficial bacteria in the second mixture to be 1-5 x 10 by resuspending10And CFU/mL, the concentration of the beneficial bacteria is controlled, the content of the beneficial bacteria in the prepared probiotic capsule is ensured, and the action effect of the probiotic capsule is ensured. Preferably, the solution subjected to resuspension treatment is selected from sterile physiological saline or sterile ultrapure water. In the preferred embodiment of the present invention, the first mixture pellet is resuspended in 2mL of sterile physiological saline to obtain the second mixture.
In the step S02, a zein solution is provided, the second mixture is added to the zein solution within 1 to 2 minutes, and the mixture is uniformly mixed to obtain a third mixture.
Preferably, the concentration of the zein solution is 8-12 mg/mL, and the concentration of the zein solution is controlled to ensure that a stable embedding effect can be formed. If the concentration of the zein solution is too low, the solution is too dilute, so that an embedding structure is not favorably formed; if the concentration of the zein solution is too high, the formed embedding structure is not uniform enough and is easy to fall off due to the excessive thickening of the solution.
Preferably, the method for preparing the zein solution comprises the following steps: providing zein particles, adding the zein particles into an ethanol solution, and performing ultrasonic mixing to obtain a zein solution; wherein the volume fraction of the ethanol solution is 75-80%.
Further, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture. And adding the second mixture into the zein solution within 1-2 minutes, controlling the second mixture to be added into the zein solution at a higher speed, and controlling the dropping speed to be higher so as to ensure that a core-shell structure is formed. If the dropping speed is too slow, the probiotic capsule with a round core-shell structure cannot be formed, so that the shape and size of the probiotic capsule are influenced, and the coating effect is also influenced.
Preferably, the mass ratio of the second mixture to the zein solution is controlled to be 1 (0.8-1.2), so that the second mixture and the zein solution can well form a compound with a core-shell structure.
Preferably, the mixing method includes, but is not limited to, mixing by vortex oscillation, in order to ensure good mixing effect and uniform mixing.
Specifically, in step S03, freeze-drying the third mixture to obtain a probiotic capsule; wherein, the freeze drying method is convenient and rapid, which is favorable for rapidly obtaining the probiotic capsule
Specifically, the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein. Preferably, the particle size of the core is 1.5 to μm, and the thickness of the shell is 0.5 μm. The prepared probiotic capsule comprises an inner core and a shell for coating the inner core, so that a core-shell structure is formed, and furthermore, the probiotic is used as an inner core material, so that the probiotic is protected by the shell material in the capsule preparation and acting on the large intestine, and the tolerance of the probiotic is improved; the shell material comprises a probiotic protective agent and a zein solution, the probiotic protective agent is used as the shell material of the probiotic capsule, the performance of the probiotics is further protected, the probiotics keeps good stability in the preparation process of the probiotic capsule, and meanwhile, the tolerance of the probiotics during the operation in intestines and stomach is ensured to be improved, and the probiotics cannot be inactivated due to the acidic environment of the stomach; secondly, zein is used as a shell material of the probiotic capsule, the zein is a hydrophobic macromolecule, has good biocompatibility and degradability, can be used as a carrier of probiotics to embed the probiotics, and is rich in sulfur amino acid, and protein molecules are connected by strong disulfide bonds and hydrophobic bonds, so that the shell structure of the prepared probiotic capsule is smooth and flat, and meanwhile, the zein has low permeability to oxygen, can effectively prevent the medicine from contacting with external oxygen, improves the storage time of the core material probiotics, and simultaneously ensures that the tolerance is improved when the capsule works in intestines and stomach, and the capsule cannot be inactivated due to the acidic environment of the stomach, thereby ensuring the release of the probiotic capsule in the large intestine and improving the effect of the probiotics.
The following will explain the present invention by way of specific examples.
Example 1
Probiotics capsule and preparation method thereof
The probiotic capsule is prepared by the following preparation method:
providing a rhamnosus bacterium suspension as a probiotic solution, providing skim milk powder, trehalose and glycerol as probiotic protective agents;
mixing a probiotic solution and a probiotic protective agent to obtain a first mixture, adding a hydrogen potassium phthalate solution dropwise to adjust the pH of the first mixture to 4.0, standing for 30 minutes, centrifuging at 4 ℃ and 1000rpm for 30 minutes, collecting a first mixture precipitate, washing with sterile normal saline for three times, carrying out heavy suspension treatment on the first mixture precipitate with sterile normal saline to obtain a second mixture, and controlling the bacterial liquid concentration of the second mixture to be 1-5 × 1010CFU/mL；
Providing zein particles, adding the zein particles into an 80% ethanol solution, performing ultrasonic mixing for 10 minutes to obtain a zein solution with the concentration of 8-12 mg/mL, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain a probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein; the grain diameter of the inner core is 1.5 μm, and the thickness of the outer shell is 0.5 μm; the mass ratio of the shell to the core is 3: 1.
example 2
Probiotics capsule and preparation method thereof
The probiotic capsule is prepared by the following preparation method:
providing a rhamnosus bacterium suspension as a probiotic solution, providing skim milk powder, trehalose and glycerol as probiotic protective agents;
mixing a probiotic solution and a probiotic protective agent to obtain a first mixture, adding a hydrogen potassium phthalate solution dropwise to adjust the pH of the first mixture to 4.0, standing for 30 minutes, then performing centrifugal treatment at 4 ℃ and 1000rpm for 30 minutes, collecting a first mixture precipitate, cleaning the first mixture precipitate with sterile normal saline for three times, performing heavy suspension treatment on the first mixture precipitate with sterile normal saline to obtain a second mixture, and controlling the concentration of a bacterium solution of the second mixture to be 1-5 × 1010 CFU/mL;
providing zein particles, adding the zein particles into an 80% ethanol solution, performing ultrasonic mixing for 10 minutes to obtain a zein solution with the concentration of 8-12 mg/mL, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain a probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein; the grain diameter of the inner core is 1.5 μm, and the thickness of the outer shell is 0.5 μm; the mass ratio of the shell to the core is 2: 1.
example 3
Probiotics capsule and preparation method thereof
The probiotic capsule is prepared by the following preparation method:
providing a rhamnosus bacterium suspension as a probiotic solution, providing skim milk powder, trehalose and glycerol as probiotic protective agents;
mixing a probiotic solution and a probiotic protective agent to obtain a first mixture, adding a hydrogen potassium phthalate solution dropwise to adjust the pH of the first mixture to 4.0, standing for 30 minutes, then performing centrifugal treatment at 4 ℃ and 1000rpm for 30 minutes, collecting a first mixture precipitate, cleaning the first mixture precipitate with sterile normal saline for three times, performing heavy suspension treatment on the first mixture precipitate with sterile normal saline to obtain a second mixture, and controlling the concentration of a bacterium solution of the second mixture to be 1-5 × 1010 CFU/mL;
providing zein particles, adding the zein particles into an 80% ethanol solution, performing ultrasonic mixing for 10 minutes to obtain a zein solution with the concentration of 8-12 mg/mL, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain a probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein; the grain diameter of the inner core is 1.5 μm, and the thickness of the outer shell is 0.5 μm; the mass ratio of the shell to the core is 1: 3.
example 4
Probiotics capsule and preparation method thereof
The probiotic capsule is prepared by the following preparation method:
providing a rhamnosus bacterium suspension as a probiotic solution, providing skim milk powder, trehalose and glycerol as probiotic protective agents;
mixing the probiotic solution and the probiotic protective agent to obtain a first mixture, and adding dropwise potassium hydrogen phthalate solution to adjust the first mixtureThe pH value is 4.0, the mixture is kept stand for 30 minutes, then centrifugal treatment is carried out for 30 minutes under the conditions of 4 ℃ and 1000rpm, the first mixture precipitate is collected and washed with sterile normal saline three times, the first mixture precipitate is subjected to heavy suspension treatment by using the sterile normal saline to obtain a second mixture, and the concentration of bacterial liquid of the second mixture is controlled to be 1-5 multiplied by 1010CFU/mL；
Providing zein particles, adding the zein particles into an 80% ethanol solution, performing ultrasonic mixing for 10 minutes to obtain a zein solution with the concentration of 8-12 mg/mL, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain a probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein; the grain diameter of the inner core is 1.5 μm, and the thickness of the outer shell is 0.5 μm; the mass ratio of the shell to the core is 1: 2.
example 5
Probiotics capsule and preparation method thereof
The probiotic capsule is prepared by the following preparation method:
providing a lactobacillus suspension as a probiotic solution, and providing skim milk powder, trehalose and glycerol as probiotic protective agents;
mixing a probiotic solution and a probiotic protective agent to obtain a first mixture, adding a hydrogen potassium phthalate solution dropwise to adjust the pH of the first mixture to 4.0, standing for 30 minutes, centrifuging at 4 ℃ and 1000rpm for 30 minutes, collecting a first mixture precipitate, washing with sterile normal saline for three times, carrying out heavy suspension treatment on the first mixture precipitate with sterile normal saline to obtain a second mixture, and controlling the bacterial liquid concentration of the second mixture to be 1-5 × 1010CFU/mL；
Providing zein particles, adding the zein particles into an 80% ethanol solution, performing ultrasonic mixing for 10 minutes to obtain a zein solution with the concentration of 8-12 mg/mL, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain a probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein; the grain diameter of the inner core is 1.5 μm, and the thickness of the outer shell is 0.5 μm; the mass ratio of the shell to the core is 2: 1.
example 6
Probiotics capsule and preparation method thereof
The probiotic capsule is prepared by the following preparation method:
providing a lactobacillus suspension as a probiotic solution, and providing skim milk powder, trehalose and glycerol as probiotic protective agents;
mixing a probiotic solution and a probiotic protective agent to obtain a first mixture, adding a hydrogen potassium phthalate solution dropwise to adjust the pH of the first mixture to 4.0, standing for 30 minutes, centrifuging at 4 ℃ and 1000rpm for 30 minutes, collecting a first mixture precipitate, washing with sterile normal saline for three times, carrying out heavy suspension treatment on the first mixture precipitate with sterile normal saline to obtain a second mixture, and controlling the bacterial liquid concentration of the second mixture to be 1-5 × 1010CFU/mL；
Providing zein particles, adding the zein particles into an 80% ethanol solution, performing ultrasonic mixing for 10 minutes to obtain a zein solution with the concentration of 8-12 mg/mL, adding the zein solution into the second mixture within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain a probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of a probiotic bacteria protective agent and zein; the grain diameter of the inner core is 1.5 μm, and the thickness of the outer shell is 0.5 μm; the mass ratio of the shell to the core is 2: 1.
analysis of results
EXAMPLES 1E to E4, simulating the release process of the prepared probiotic capsule in vitro, respectively placing the probiotic capsule prepared in the embodiment 1-4 in artificial gastric juice for treatment, wherein the treatment time is not more than 2 hours, then placing the probiotic capsule in artificial small intestine juice for treatment, wherein the treatment time is not more than half 4 hours, and then placing the probiotic capsule in artificial large intestine juice for treatment; the in vitro release of probiotics in the probiotic capsules prepared in the different examples was further analyzed. As shown in the attached figure 2, the probiotic capsule of the example 1 (the mass ratio of the outer shell to the inner core is 3: 1) releases the most beneficial bacteria, and the coating amount of the beneficial bacteria is 2.47 x 109CFU/mL; the probiotic capsule of example 2 (mass ratio of shell to core of 2: 1) released the most beneficial bacteria, with a beneficial bacteria coating of 2.50 × 109CFU/mL; the probiotic capsule of example 3 (mass ratio of shell to core 1: 3) released the most beneficial bacteria, with a beneficial bacteria coating of 5.3 × 108CFU/mL; the probiotic capsule of example 4 (mass ratio of shell to core of 1: 2) released the most beneficial bacteria, and the coating amount of beneficial bacteria was 1.52 × 109CFU/mL. It can be seen that the mass ratio of shell to core prepared from example 2 was 2: the content of probiotics coated by the probiotic capsule of 1 is the most, and the structure of the probiotic capsule comprises an inner core and a shell coating the inner core, so that a core-shell structure is formed, the probiotics are protected by a shell material in the capsule preparation and acting on the large intestine, and the tolerance of the probiotics is improved.
The present invention is not limited to the above embodiments, and any modification, equivalent replacement, and improvement made within the spirit and principle of the embodiments should be included in the protection scope of the embodiments.
Claims (10)
1. A probiotic capsule, characterized in that it comprises an inner core and an outer shell covering said inner core; wherein, the material of the inner core comprises probiotics, and the material of the outer shell comprises a probiotic protective agent and zein.
2. The probiotic capsule according to claim 1, characterized in that the mass ratio of the outer shell to the inner core is (2-3): 1.
3. The probiotic capsule according to claim 1, characterized in that the particle size of the inner core is 1.5 to 1.8 μm and the thickness of the outer shell is 0.5 to 0.8 μm.
4. The probiotic capsule according to any one of claims 1 to 3, characterized in that the probiotic protecting agent comprises the following components in parts by weight:
5. the preparation method of the probiotic capsule is characterized by comprising the following steps:
mixing a probiotic solution and a probiotic protective agent to obtain a first mixture, adjusting the pH of the first mixture to 3.8-4.5, standing, performing centrifugal treatment, collecting a first mixture precipitate, and performing heavy suspension treatment on the first mixture precipitate to obtain a second mixture;
providing a zein solution, adding the second mixture into the zein solution within 1-2 minutes, and uniformly mixing to obtain a third mixture;
freeze-drying the third mixture to obtain the probiotic capsule; the probiotic capsule comprises an inner core and a shell coating the inner core, wherein the inner core is made of probiotic bacteria, and the shell is made of probiotic bacteria protective agent and zein.
6. The method for preparing a probiotic capsule according to claim 5, wherein in the step of mixing a probiotic solution and a probiotic protecting agent to obtain a first mixed solution and adjusting the pH of the first mixture to 3.8 to 4.5, the first mixed solution is added dropwise to a buffer solution to adjust the pH of the first mixture to 3.8 to 4.5; wherein the buffer solution is at least one selected from potassium hydrogen phthalate solution, disodium hydrogen phosphate-citric acid buffer solution and citric acid-sodium citrate buffer solution.
7. The method for preparing probiotic capsule according to claim 6, wherein the concentration of probiotic in the first mixture is 1-9 x 1010CFU/mL。
8. The method for preparing the probiotic capsule according to claim 6, wherein the mass ratio of the probiotics to the buffer solution in the first mixed solution is 1: (2-2.5); and in the step of dropwise adding the first mixed solution into the buffer solution, the dropwise adding speed is 1-1.5 drops/second.
9. The method of preparing a probiotic capsule according to claim 5, wherein the method of preparing the zein solution comprises the steps of: providing zein particles, adding the zein particles into an ethanol solution, and performing ultrasonic mixing to obtain the zein solution; wherein the volume fraction of the ethanol solution is 75-80%.
10. The method for preparing probiotic capsules according to claim 5, wherein the concentration of beneficial bacteria in the second mixture is 1 to 5 x 1010CFU/mL; and/or the presence of a gas in the gas,
the concentration of the zein solution is 8-12 mg/mL.
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