CN117297100B - Probiotic microcapsule prepared by multifluid coaxial spray and preparation method and application thereof - Google Patents

Probiotic microcapsule prepared by multifluid coaxial spray and preparation method and application thereof Download PDF

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CN117297100B
CN117297100B CN202311597217.0A CN202311597217A CN117297100B CN 117297100 B CN117297100 B CN 117297100B CN 202311597217 A CN202311597217 A CN 202311597217A CN 117297100 B CN117297100 B CN 117297100B
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oil
probiotic
lipid
core material
preparation
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CN117297100A (en
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谢有发
邹立强
张奎
刘颖
刘伟
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Nanchang University
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Nanchang University
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/015—Inorganic compounds
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    • A23L29/04—Fatty acids or derivatives
    • A—HUMAN NECESSITIES
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    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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    • A—HUMAN NECESSITIES
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    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/212—Starch; Modified starch; Starch derivatives, e.g. esters or ethers
    • A—HUMAN NECESSITIES
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    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/231—Pectin; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/238—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin from seeds, e.g. locust bean gum or guar gum
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/256—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin from seaweeds, e.g. alginates, agar or carrageenan
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/269—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of microbial origin, e.g. xanthan or dextran
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/269—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of microbial origin, e.g. xanthan or dextran
    • A23L29/272—Gellan
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/275—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of animal origin, e.g. chitin
    • A23L29/281—Proteins, e.g. gelatin or collagen
    • A23L29/284—Gelatin; Collagen
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • A—HUMAN NECESSITIES
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    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • A23L29/35—Degradation products of starch, e.g. hydrolysates, dextrins; Enzymatically modified starches
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    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • A23L29/37—Sugar alcohols
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P20/00—Coating of foodstuffs; Coatings therefor; Making laminated, multi-layered, stuffed or hollow foodstuffs
    • A23P20/20—Making of laminated, multi-layered, stuffed or hollow foodstuffs, e.g. by wrapping in preformed edible dough sheets or in edible food containers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D1/00—Evaporating
    • B01D1/16—Evaporating by spraying
    • B01D1/18—Evaporating by spraying to obtain dry solids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D1/00—Evaporating
    • B01D1/16—Evaporating by spraying
    • B01D1/20—Sprayers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02—Making microcapsules or microballoons
    • B01J13/04—Making microcapsules or microballoons by physical processes, e.g. drying, spraying
    • B01J13/043—Drying and spraying
    • A—HUMAN NECESSITIES
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    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract

The invention discloses a probiotic microcapsule prepared by multifluid coaxial spraying, a preparation method and application thereof. The probiotic microcapsule of the invention is formed by adding an intermediate emulsified lipid layer between a core material and a wall material. The design of the emulsified lipid layer can play a role in absorbing heat and reduce the thermal death of the probiotics in the inner layer; meanwhile, a natural physical barrier can be formed, and the gastrointestinal tract protection effect is enhanced; the crosslinking factors can be suspended and released to crosslink with the gel on the outer layer, so that the capsule has more stable structure and better gastrointestinal protection effect. The three-layer microcapsule is prepared by a novel multi-fluid coaxial spray drying device, and the device is provided with a four-fluid coaxial nozzle based on the existing coaxial spray device, so that the three-layer microcapsule is formed.

Description

Probiotic microcapsule prepared by multifluid coaxial spray and preparation method and application thereof
Technical Field
The invention relates to the technical field of microbial preparations, in particular to a probiotic microcapsule prepared by multifluid coaxial spraying, and a preparation method and application thereof.
Background
Probiotics are living microorganisms which can produce beneficial effects on host health after being ingested in certain amounts, and play an important role in regulating intestinal flora, inhibiting pathogenic bacteria, regulating immunity, reducing blood sugar and blood fat, preventing diseases and the like. The probiotic effect of probiotic products is largely dependent on the number of viable bacteria colonized the colon, but probiotics are susceptible to environmental stresses (including high temperature, humidity, oxygen stress, osmotic pressure) during food processing, storage, low pH and bile salts during digestion leading to cell damage and even loss of viability. In order to reduce the loss of activity of probiotics under these environmental stresses, the use of microencapsulation techniques to prepare oral delivery systems for probiotics is a very effective approach.
The probiotics microcapsule is prepared by encapsulating probiotics in a protective wall material through a microcapsule technology, so that the resistance of the probiotics to various stress factors in the environment can be effectively improved, the shelf life is prolonged, and the targeted controlled release in the intestinal tract is realized. Spray drying is a microencapsulation technology suitable for industrialization, and by the combined action of atomization, heating and drying, the dry powder is rapidly generated in one step, and the method has the advantages of high efficiency and economy. The prior art reports that lactobacillus plantarum LP115 is subjected to microencapsulation by a spray drying technology, and the result shows that a large amount of probiotics still survive after the microcapsule with the optimized formula is stored for 12 months at room temperature. The prior art also reports that the preparation of sodium alginate-whey protein isolate microcapsules by an emulsification method and embedding of lactobacillus acidophilus are also improved in bacterial activity after digestion of the encapsulated probiotics compared with free probiotics. However, during spray drying, the probiotic cells may be subjected to heat stress, dehydration, oxygen exposure and osmotic stress, resulting in damage to membranes, proteins, DNA and RNA. The prior art also found that bifidobacterium infantis CCRC 10 survived only 2.15% when spray-dried in 10% (w/w) acacia gum. Also, there is a report that 0.70 log CFU/mL of kefir 8348 lactic acid bacteria were lost during spray drying in 11% (w/v) of recombinant skim milk. Similar results for varying degrees of cell death during spray drying have also been reported by other researchers.
Therefore, there is a need to develop a better spray drying process to increase the probiotic viability during spray drying and to obtain probiotic microcapsules that are gastrointestinal tract protective and long-term storage.
In view of this, the present invention has been made.
Disclosure of Invention
The invention aims to provide a probiotic microcapsule prepared by multifluid coaxial spraying and a preparation method and application thereof.
The invention is realized in the following way:
in a first aspect, the present invention provides a probiotic microcapsule comprising a core material and a wall material, and an emulsified lipid layer surrounding the core material.
In a second aspect, the invention provides a method for preparing the probiotic microcapsules, which comprises preparing the raw materials of the core material, the wall material and the emulsified lipid layer into the probiotic microcapsules by adopting a multi-fluid coaxial spray drying device.
In a third aspect, the invention also provides the use of the probiotic microcapsules described above in the preparation of an oral probiotic preparation.
The invention has the following beneficial effects:
The probiotic microcapsule of the invention adds an intermediate emulsified lipid layer between the core material and the wall material, so that the probiotic microcapsule forms a multi-layer microcapsule. The design of the emulsified lipid layer can play a role in absorbing heat and reduce the thermal death of the probiotics in the inner layer; meanwhile, a natural physical barrier can be formed, and the gastrointestinal tract protection effect is enhanced; the crosslinking factors can be suspended and released to crosslink with the gel on the outer layer, so that the capsule has more stable structure and better gastrointestinal protection effect. The multilayer microcapsule is prepared by a novel multi-fluid coaxial spray drying device, and the device is provided with a four-fluid coaxial nozzle based on the existing coaxial spray device, so that the three-layer microcapsule is formed.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic illustration of a four-fluid coaxial nozzle in a multiple-fluid coaxial spray drying apparatus employed in the present invention; wherein 1 is an inner layer material inlet; 2-an inner layer material outlet; 3-an intermediate material inlet; 4-an intermediate material outlet; 5-an outer layer material inlet; 6-an outer layer material outlet; 7-a spray air inlet; 8-air flow channels;
Fig. 2 is a schematic structural diagram of the probiotic microcapsules prepared according to the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
A probiotic microcapsule comprises a core material, an emulsified lipid layer wrapping the core material, and a wall material wrapping the emulsified lipid layer.
The probiotic microcapsule is prepared by mixing an emulsifying agent, lipid and a cross-linking agent, wherein the lipid can be one or more of corn oil, medium Chain Triglyceride (MCT), soybean oil, sunflower seed oil, peanut oil, sesame oil, rice bran oil, canola oil, rapeseed oil, walnut oil, olive oil, hydrogenated vegetable oil, cocoa butter replacer, palm oil, beeswax, stearic acid, lard, butter, beef tallow and coconut oil.
In the invention, the emulsifier comprises one or more of phospholipid, mono-diglyceride fatty acid ester, esterified sorbitol fatty acid ester, sucrose fatty acid ester, polyglycerol fatty acid ester, sorbitan monostearate and polyglycerol ricinoleate.
In the present invention, the emulsifier is added to help form a three-layer structure when sprayed.
The amount of the emulsifier added has an important influence on the formation of the emulsified lipid layer, and if the amount of the emulsifier added is too low, the intermediate lipid layer of the microcapsule leaks, a three-layer structure cannot be formed, and if the amount of the emulsifier added is too high, the viscosity of the lipid layer is too high, and spraying cannot be performed during preparation. On the basis, the invention carries out further screening test on the addition amount of the emulsifier, and discovers that when the mass ratio of the emulsifier to the lipid is 1-20:80-99, and can obtain better effect.
In the present invention, the crosslinking agent includes one or more of calcium chloride, calcium lactate and potassium chloride, and more preferably, the crosslinking agent is calcium chloride. The cross-linking agent is added into the emulsified lipid layer, and the cross-linking agent and the lipid suspension cross-linking factor can be released to cross-link with the wall material of the outer layer to form a denser network structure, so that the microcapsule structure is more stable.
The addition amount of the cross-linking agent has an important influence on the preparation of the microcapsule, if the content is too high, the spray nozzle of the spraying equipment is easy to be blocked during the preparation, and if the content is too low, the cross-linking effect cannot be brought, so that the influence on the improvement of structural stability is small. On the basis, the invention carries out further screening test on the addition amount of the cross-linking agent, and discovers that when the mass ratio of the emulsifying agent to the cross-linking agent to the lipid is 1-20:0.5-3:80-99, and can obtain better effect.
The emulsified lipid layer prepared by the emulsifier, the lipid and the cross-linking agent can play a role in absorbing heat in the preparation process, and reduce the thermal death of inner probiotics; a natural physical barrier is formed during digestion to enhance the gastrointestinal tract protecting effect.
The core material wrapped by the emulsified lipid layer comprises probiotics and prebiotics. The probiotics can be one or more of lactobacillus plantarum, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus casei, lactobacillus paracasei, lactobacillus rhamnosus, lactobacillus reuteri, bifidobacterium animalis subspecies lactis, bifidobacterium adolescentis subspecies longum, bifidobacterium breve and mannitzerland bacteria, and can also be other probiotics or probiotic combinations.
The prebiotic comprises one or more of fructo-oligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide and inulin. The prebiotics are added into the core material to provide nutrition for the probiotics, selectively stimulate the growth and activate metabolism of the probiotics, and endow the probiotics with competitive advantages in intestinal tracts, thereby beneficially affecting the host.
The wall material for wrapping the emulsified lipid layer comprises water, a gel agent and a filler. The gel comprises one or more of gelatin, pectin, gellan gum, sodium alginate, carrageenan, locust bean gum, agar, xanthan gum, guar gum, tragacanth gum, starch, etc. The addition amount is controlled to be within 400 mPa.s of the viscosity of the solution of the wall material, and preferably, the addition amount of the gel agent is 1-10% of the mass of water.
The filler comprises one or more of maltodextrin, sucrose, cyclodextrin, erythritol, xylitol and acacia, and preferably the filler is added in an amount of 10-30% of the mass of water.
The combination of the core material, the emulsified lipid layer and the wall material jointly form a microcapsule with a three-layer structure, and more sufficient protection effect is provided for probiotics in the core material through the synergistic effect of the emulsified lipid layer and the wall material.
Correspondingly, the invention can obtain microcapsules with a three-layer structure, and also depends on a novel coaxial spray drying arrangement, wherein the device comprises a four-fluid coaxial nozzle, a drying tower and a collector which are communicated in sequence;
As shown in fig. 1, the four-fluid coaxial nozzle is designed according to the microcapsule of the present invention, and includes an outer layer material inlet 5, an outer layer material outlet 6, an intermediate material inlet 3, an intermediate material outlet 4, an inner layer material inlet 1, an inner layer material outlet 2, a spray air inlet 7, and an air flow passage 8. Specifically, be equipped with inlayer material import 1 on the top of four fluid coaxial nozzle, the lateral wall is equipped with spraying air inlet 7, outer material import 5 and middle material import 3 respectively, inside be equipped with respectively with inlayer material export 2, the middle material export 4 with middle material import 3 intercommunication, outer material export 6 with outer material import 5 intercommunication, the air current channel 8 with spraying air inlet 7 intercommunication with inlayer material export 2, middle material export 4 and outer material export 6 coaxial setting. The drying tower and collector are conventional devices, as the invention is not limited in this regard.
When the microcapsule drying device is used, corresponding materials are respectively added into the inner material inlet 1, the middle material inlet 3 and the outer material inlet 5, the forming of the microcapsule is realized by regulating and controlling the relevant parameters of the device, the formed microcapsule flows into a drying tower communicated with the four-fluid coaxial nozzle, the drying of the microcapsule is realized under the action of hot air flow, and then the microcapsule enters a collector, so that the microcapsule finished product is obtained.
Specifically, the preparation method of the microcapsule comprises the following steps:
s1, preparing wall material: adding the gel into water, stirring at 60-70 ℃ until the gel is completely dissolved, and then adding the filler.
In the wall material, the addition amount of the gel is 1-10% of the mass of water, and the addition amount of the filler is 10-30% of the mass of water.
S2, preparing an emulsified lipid layer material: adding the emulsifying agent into the lipid, heating and slowly stirring until the emulsifying agent is completely dissolved and the lipid is completely melted to prepare an emulsified lipid layer, mixing the cross-linking agent into the emulsified lipid layer, and fully and uniformly stirring the cross-linking agent for later use.
In the preparation process, the mass ratio of the emulsifier, the cross-linking agent and the lipid in the emulsified lipid layer is 1-20: 0.5-3: 80-99.
S3, preparing core materials: and adding the probiotic bacteria powder and the prebiotics into deionized water, and stirring to dissolve and uniformly mix the probiotic bacteria powder and the prebiotics.
The addition amount of the probiotics powder in the core material is 1-20% of the mass of the water, and the addition amount of the probiotics is 1-10% of the mass of the water.
S4, respectively introducing the core material, the emulsified lipid layer material and the wall material into the inner, middle and outer feed channels of the four-fluid coaxial nozzle, and collecting the probiotic microcapsules.
Wherein the air inlet temperature of the multi-fluid coaxial spray drying equipment is 100-130 ℃; the temperature of the air outlet is 60-80 ℃.
When the four-fluid coaxial nozzle is introduced into the inner, middle and outer feed channels of the four-fluid coaxial nozzle, the volume ratio of the core material to the emulsified lipid layer material to the wall material is 1-2:2-5:10-50.
The features and capabilities of the present invention are described in further detail below in connection with the examples.
Example 1
The embodiment provides a preparation method of a probiotic microcapsule, which comprises the following steps:
(1) Preparing a wall material: 1 part of gellan gum and 4 parts of gelatin are added to 75 parts of deionized water, 5 parts of maltodextrin and 15 parts of sucrose are added, and stirred at 60 ℃ until completely dissolved.
(2) Preparation of an emulsified lipid layer: 10 parts of soybean phospholipids were added to 90 parts of MCT, heated and stirred slowly until the soybean phospholipids were completely dissolved, cooled, and 1 part of calcium chloride was mixed therein and dispersed using a dispersing machine for use.
(3) Preparing an inner layer probiotics liquid: adding 10 parts of lactobacillus plantarum P9 bacterial powder (from the river Chinese medicine industry) into 85 parts of deionized water stock limited) and 5 parts fructooligosaccharides were dissolved and mixed well with stirring.
(4) The formation and drying of the multilayer microcapsules is achieved by means of a Buchi-290 spray drying tower equipped with a four-fluid coaxial nozzle. The temperature of the inlet air is regulated to be constant at 120 ℃, the mixed solution of probiotics, emulsified lipid and wall material solution are respectively introduced into the inner, middle and outer feeding channels of the four-fluid coaxial nozzle, and the probiotic microcapsules at the outlet air temperature of 70 ℃ are collected, and the prepared microcapsules are shown in figure 2.
Example 2
This example provides a method of preparing probiotic microcapsules, which differs from example 1 in the formulation of the emulsified lipid layer: 10 parts of soybean lecithin is added into 90 parts of coconut oil, heated and slowly stirred until the soybean lecithin is completely dissolved, the coconut oil is melted, and calcium chloride is mixed into the mixture and fully dispersed for standby.
Example 3
The present example provides a method for preparing probiotic microcapsules, which differs from example 1 in the formulation of the wall material: 2 parts of sodium alginate and 3 parts of gelatin are added to 75 parts of deionized water, then 5 parts of maltodextrin and 15 parts of sucrose are added, and stirred at 60 ℃ until completely dissolved.
Example 4
This example provides a method of preparing probiotic microcapsules, which differs from example 1 in the formulation of the emulsified lipid layer: 10 parts of mono-di-glycerin fatty acid ester is added into 90 parts of MCT, heated and slowly stirred until the mono-di-glycerin fatty acid ester is completely dissolved, the MCT is melted, and calcium chloride is mixed into the MCT and fully dispersed for standby.
Comparative example 1
This comparative example differs from example 1 in the formulation of the intermediate layer: no emulsifier and lipid were added and only 1% (w/w) CaCl 2 was added to deionized water and dissolved.
Comparative example 2
The difference between this comparative example and example 1 is the formulation of the emulsified lipid layer: no emulsifying agent is added.
In the preparation of the emulsified lipid layer, it was found that CaCl 2 could not be suspended without adding an emulsifier, but CaCl 2 could be suspended in the emulsified lipid layer obtained by the preparation method of the present invention.
Comparative example 3
The comparative example differs from example 1 in that the amount of soybean phospholipids added to the emulsified lipid layer was 0.5 parts and the amount of MCT added was 99.5 parts.
Comparative example 4
This comparative example differs from example 1 in that no emulsified lipid layer was provided.
Comparative example 5
This comparative example differs from example 1 in that the emulsified lipid layer does not contain CaCl 2.
Specific materials for examples 1 to 4 and comparative examples 1 to 5 are shown in Table 1:
table 1 specific materials for different examples and comparative examples
Experimental example 1
The conditions of each example and comparative example at the time of preparing microcapsules and the collected microcapsules were recorded, and the spraying conditions of each microcapsule are shown in table 2:
TABLE 2 spray conditions of different microcapsules
Examples Spray conditions
Example 1 Good quality
Example 2 Good quality
Example 3 Good quality
Example 4 Good quality
Comparative example 1 Good quality
Comparative example 2 Difficult to embed and has the phenomenon of oil leakage
Comparative example 3 Difficult to embed and has the phenomenon of oil leakage
Comparative example 4 Good quality
Comparative example 5 Good quality
From the spray condition of the different microcapsules of table 2, the probiotic microcapsules obtained by the preparation method of the present invention can form a stable three-layer structure, and the problem of leakage of the lipid layer in the middle of the microcapsules does not occur, compared with comparative example 2 and comparative example 3.
Experimental example 2
The experimental example detects the activity of probiotics in the preparation process, the simulated digestion process and the storage process respectively:
(1) Probiotics vitality in preparation process
0.1G of the microcapsule was taken in 9.9 mL of a 0.5% sodium citrate solution, incubated at 37℃for 10 min, vortexed until the microcapsule was completely dissolved, the solution was serially diluted with physiological saline (0.9% sodium chloride) and then applied to MRS agar medium, and after aerobic culture at 37℃for 48: 48 h, colony counts were performed, and the viable count at the subsequent storage and digestion was determined by the same method.
TABLE 3 Probiotics changes during preparation
As can be seen from the data in table 3, the present invention can significantly improve the survival rate of probiotics during capsule preparation by providing an emulsified lipid layer, which may be due to less heat loss of probiotics during spraying caused by the heat absorption of the lipid.
(2) Probiotic viability in simulated digestion
Configuration of simulated gastric digest (SGF): pepsin was added at 3.2 mg/mL to a mixed solution containing 2. 2 mg/mL NaCl and 7. 7 mL/L HCl and the pH was adjusted to 2.5. Simulated intestinal digestive fluid (SIF) configuration: 36.7 g calcium chloride, 218.7 g sodium chloride were weighed, dissolved in distilled water and set to a volume of 1L.
1G of the microcapsules was added to 9 mL SGF and placed in a water bath at 37℃under 100 rpm with shaking 2h, after gastric digestion the pH of the mixture was adjusted to 7.0 with 0.25M NaOH. 1.5 mL simulated intestinal digests, 189 mg bovine bile salts (dissolved in 3.5 mL 5mM PBS buffer, pH=7.0), 60mg lipase and 60mg trypsin (dissolved in 2.5 mL 5mM PBS buffer, pH=7.0) were added and the mixture was kept under water bath shaking at 37℃and 100 rpm for 2 h. Taking digestive juice after digestion of stomach and intestines respectively, adding 0.5% sodium citrate solution for dissolution, diluting with physiological saline, coating on MRS culture medium, aerobically culturing at 37 ℃ for 48 h, and performing colony counting.
TABLE 4 simulation of probiotic viability changes during digestion
As can be seen from Table 4, caCl 2, which was used in comparative example 1 and comparative examples 4 to 5, was effective in improving the activity of the microcapsules when digested. Meanwhile, the emulsified lipid layer also improves the protection effect of the microcapsule on probiotics during simulated digestion.
(3) Probiotics viability during storage
The microcapsules and the drying agent are sealed in an aluminum foil bag and stored in an environment of 4 ℃ and 25 ℃, sampling is carried out every 3 months, and the number of viable bacteria in the microcapsules is measured by a plate counting method.
TABLE 5 Probiotics viability Change during storage
TABLE 6 Probiotics survival during storage
From tables 5 and 6 it can be seen that the addition of an emulsified lipid layer can increase the survival rate of probiotics in the microcapsules encapsulating the probiotics during storage, thanks to the oxygen-barrier and water-barrier properties of the lipid layer.
Experimental example 3
The microcapsules of example 1 and comparative example 5 were dissolved in water as follows: comparative example 5, without calcium chloride added, was dissolved in water, while the microcapsules of example 1 were in an insoluble state in water, indicating successful crosslinking of the calcium chloride with the outer gel. This shows that the emulsified fat can release suspended calcium chloride to the outer layer wall material and react with the outer layer to reduce the water solubility of the wall material, so that the microcapsule structure is more stable and the gastrointestinal protection effect is better.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (5)

1. The probiotic microcapsule is characterized by comprising a core material and a wall material, wherein the core material contains probiotics, and the probiotic microcapsule also comprises an emulsified lipid layer wrapping the core material; the emulsified lipid layer comprises an emulsifier, lipid and a cross-linking agent;
the mass ratio of the emulsifying agent, the cross-linking agent and the lipid in the emulsified lipid layer is 1-20:0.5-3:80-99;
The emulsifier is one or more of phospholipid, mono-diglyceride fatty acid ester, esterified sorbitol fatty acid ester, sucrose fatty acid ester, polyglycerol fatty acid ester, sorbitan monostearate and polyglycerol polyricinoleate;
The lipid is one or more selected from corn oil, medium chain triglyceride, soybean oil, sunflower seed oil, peanut oil, sesame oil, rice bran oil, canola oil, rapeseed oil, walnut oil, olive oil, hydrogenated vegetable oil, cocoa butter replacer, palm oil, beeswax, stearic acid, lard, butter, beef tallow, and coconut oil;
the cross-linking agent is one or more selected from calcium chloride, calcium lactate and potassium chloride;
the core material also comprises a prebiotic;
The wall material comprises a gel and a filler;
the gel is one or more of gelatin, pectin, gellan gum, sodium alginate, carrageenan, locust bean gum, agar, xanthan gum, guar gum, tragacanth gum and starch;
The filler comprises one or more of maltodextrin, sucrose, cyclodextrin, erythritol, xylitol and acacia;
The preparation method of the probiotic microcapsule comprises the following steps: preparing the raw materials of the core material, the wall material and the emulsified lipid layer into the probiotic microcapsule by adopting a multi-fluid coaxial spray drying device;
the multi-fluid coaxial spray drying equipment comprises a four-fluid coaxial nozzle, a drying tower and a collector which are sequentially communicated;
Respectively introducing the materials of the core material, the emulsified lipid layer and the wall material into an inner layer material inlet, a middle material inlet and an outer layer material inlet of the four-fluid coaxial nozzle, and completing the forming and drying of the probiotic microcapsules through the multi-fluid coaxial spray drying equipment;
In the forming and drying process, the air inlet temperature of the multi-fluid coaxial spray drying equipment is 100-130 ℃; the air outlet temperature is 60-80 ℃;
The preparation method of the core material comprises the steps of adding probiotic bacteria powder and prebiotics into water, and uniformly mixing to obtain the core material, wherein the addition amount of the probiotic bacteria powder in the core material is 1-20% of the mass of the water, and the addition amount of the prebiotics is 1-10% of the mass of the water;
The preparation method of the emulsified lipid layer material comprises adding an emulsifier into lipid, heating and stirring until the emulsifier is completely dissolved and the lipid is completely melted, and then adding a crosslinking agent and uniformly mixing; wherein, the mass ratio of the emulsifying agent, the cross-linking agent and the lipid is 1-20:0.5-3:80-99;
The preparation method of the wall material comprises the steps of adding the gel into water, stirring at 60-70 ℃ until the gel is completely dissolved, and then adding the filler; wherein the addition amount of the gel is 1-10% of the mass of the water, and the addition amount of the filler is 10-30% of the mass of the water.
2. The probiotic microcapsule according to claim 1, characterized in that the probiotics in the core material are selected from one or several of lactobacillus plantarum, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus casei, lactobacillus paracasei, lactobacillus rhamnosus, lactobacillus reuteri, bifidobacterium animalis subspecies lactis, bifidobacterium adolescentis, bifidobacterium longum subspecies longum, bifidobacterium breve, and mannheimia coagulans;
The prebiotics are selected from one or more of fructo-oligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide and inulin.
3. A probiotic microcapsule according to claim 2, characterized in that,
The four-fluid coaxial nozzle is characterized in that an inner layer material inlet is formed in the top end of the four-fluid coaxial nozzle, an outer layer material inlet and an intermediate material inlet are formed in the side wall of the four-fluid coaxial nozzle respectively, an inner layer material outlet communicated with the inner layer material inlet, an intermediate material outlet communicated with the intermediate material inlet and an outer layer material outlet communicated with the outer layer material inlet are formed in the side wall of the four-fluid coaxial nozzle respectively, and the inner layer material outlet, the intermediate material outlet and the outer layer material outlet are coaxially arranged.
4. A probiotic microcapsule according to claim 3, characterized in that the volume ratio of the core material, emulsified lipid layer material to wall material is 1-2:2-5:10-50.
5. Use of a probiotic microcapsule according to any one of claims 1 to 4 for the preparation of an oral probiotic formulation.
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