CN113749258A - Calcium alginate liquid core microcapsule and preparation method thereof - Google Patents
Calcium alginate liquid core microcapsule and preparation method thereof Download PDFInfo
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- 239000003094 microcapsule Substances 0.000 title claims abstract description 86
- 239000007788 liquid Substances 0.000 title claims abstract description 78
- 238000002360 preparation method Methods 0.000 title claims abstract description 37
- 235000010410 calcium alginate Nutrition 0.000 title claims abstract description 34
- 239000000648 calcium alginate Substances 0.000 title claims abstract description 34
- 229960002681 calcium alginate Drugs 0.000 title claims abstract description 34
- OKHHGHGGPDJQHR-YMOPUZKJSA-L calcium;(2s,3s,4s,5s,6r)-6-[(2r,3s,4r,5s,6r)-2-carboxy-6-[(2r,3s,4r,5s,6r)-2-carboxylato-4,5,6-trihydroxyoxan-3-yl]oxy-4,5-dihydroxyoxan-3-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylate Chemical compound [Ca+2].O[C@@H]1[C@H](O)[C@H](O)O[C@@H](C([O-])=O)[C@H]1O[C@H]1[C@@H](O)[C@@H](O)[C@H](O[C@H]2[C@H]([C@@H](O)[C@H](O)[C@H](O2)C([O-])=O)O)[C@H](C(O)=O)O1 OKHHGHGGPDJQHR-YMOPUZKJSA-L 0.000 title claims abstract description 34
- 239000006041 probiotic Substances 0.000 claims abstract description 45
- 235000018291 probiotics Nutrition 0.000 claims abstract description 45
- 230000000529 probiotic effect Effects 0.000 claims abstract description 23
- 238000000034 method Methods 0.000 claims abstract description 18
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 3
- 241000186000 Bifidobacterium Species 0.000 claims abstract description 3
- 241000193171 Clostridium butyricum Species 0.000 claims abstract description 3
- 241000186660 Lactobacillus Species 0.000 claims abstract description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 3
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 3
- 238000003756 stirring Methods 0.000 claims description 114
- 239000011162 core material Substances 0.000 claims description 92
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 62
- 239000001963 growth medium Substances 0.000 claims description 59
- 238000005303 weighing Methods 0.000 claims description 48
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 46
- 230000001580 bacterial effect Effects 0.000 claims description 46
- 235000010413 sodium alginate Nutrition 0.000 claims description 46
- 239000000661 sodium alginate Substances 0.000 claims description 46
- 229940005550 sodium alginate Drugs 0.000 claims description 46
- 239000000725 suspension Substances 0.000 claims description 46
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 44
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 44
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 44
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 44
- 239000001110 calcium chloride Substances 0.000 claims description 43
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 43
- 239000008367 deionised water Substances 0.000 claims description 40
- 229910021641 deionized water Inorganic materials 0.000 claims description 40
- 238000009630 liquid culture Methods 0.000 claims description 40
- 238000007872 degassing Methods 0.000 claims description 39
- 235000010493 xanthan gum Nutrition 0.000 claims description 29
- 239000000230 xanthan gum Substances 0.000 claims description 29
- 229920001285 xanthan gum Polymers 0.000 claims description 29
- 229940082509 xanthan gum Drugs 0.000 claims description 29
- 238000001914 filtration Methods 0.000 claims description 23
- 230000001112 coagulating effect Effects 0.000 claims description 22
- 238000004140 cleaning Methods 0.000 claims description 21
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 claims description 18
- 229910001424 calcium ion Inorganic materials 0.000 claims description 18
- 150000004676 glycans Chemical class 0.000 claims description 14
- 229920001282 polysaccharide Polymers 0.000 claims description 14
- 239000005017 polysaccharide Substances 0.000 claims description 14
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 12
- 230000008719 thickening Effects 0.000 claims description 9
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 7
- 229920002674 hyaluronan Polymers 0.000 claims description 7
- 229960003160 hyaluronic acid Drugs 0.000 claims description 7
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 6
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 claims description 6
- 239000001527 calcium lactate Substances 0.000 claims description 6
- 229960002401 calcium lactate Drugs 0.000 claims description 6
- 235000011086 calcium lactate Nutrition 0.000 claims description 6
- ACYGYJFTZSAZKR-UHFFFAOYSA-J dicalcium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [Ca+2].[Ca+2].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O ACYGYJFTZSAZKR-UHFFFAOYSA-J 0.000 claims description 6
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims description 5
- 229960003563 calcium carbonate Drugs 0.000 claims description 5
- 229960002713 calcium chloride Drugs 0.000 claims description 5
- 239000001768 carboxy methyl cellulose Substances 0.000 claims description 5
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 claims description 5
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 claims description 5
- 229920002148 Gellan gum Polymers 0.000 claims description 4
- 229920000161 Locust bean gum Polymers 0.000 claims description 4
- 240000004584 Tamarindus indica Species 0.000 claims description 4
- 235000004298 Tamarindus indica Nutrition 0.000 claims description 4
- 235000010492 gellan gum Nutrition 0.000 claims description 4
- 239000000216 gellan gum Substances 0.000 claims description 4
- 235000010420 locust bean gum Nutrition 0.000 claims description 4
- 239000000711 locust bean gum Substances 0.000 claims description 4
- 229920002907 Guar gum Polymers 0.000 claims description 3
- 230000015271 coagulation Effects 0.000 claims description 3
- 238000005345 coagulation Methods 0.000 claims description 3
- 235000010417 guar gum Nutrition 0.000 claims description 3
- 239000000665 guar gum Substances 0.000 claims description 3
- 229960002154 guar gum Drugs 0.000 claims description 3
- 238000011010 flushing procedure Methods 0.000 claims description 2
- 230000004083 survival effect Effects 0.000 abstract description 22
- 238000003860 storage Methods 0.000 abstract description 10
- 238000004108 freeze drying Methods 0.000 abstract description 8
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 4
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- 230000029087 digestion Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 66
- 238000005406 washing Methods 0.000 description 21
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 19
- 238000012258 culturing Methods 0.000 description 19
- 239000002504 physiological saline solution Substances 0.000 description 19
- 239000007787 solid Substances 0.000 description 19
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- 239000011148 porous material Substances 0.000 description 17
- 230000000694 effects Effects 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 239000011324 bead Substances 0.000 description 4
- 239000003833 bile salt Substances 0.000 description 4
- 238000001035 drying Methods 0.000 description 3
- 210000004211 gastric acid Anatomy 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 2
- 206010010774 Constipation Diseases 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 206010017964 Gastrointestinal infection Diseases 0.000 description 1
- 201000010538 Lactose Intolerance Diseases 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
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- 238000005516 engineering process Methods 0.000 description 1
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- 239000012530 fluid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000007407 health benefit Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- -1 hydrogen ions Chemical class 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
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- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
-
- 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
- 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
- A23L33/14—Yeasts or derivatives thereof
-
- 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
-
- 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
- B01J13/046—Making microcapsules or microballoons by physical processes, e.g. drying, spraying combined with gelification or coagulation
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Preparation (AREA)
- Manufacturing Of Micro-Capsules (AREA)
Abstract
The invention provides a calcium alginate liquid core microcapsule and a preparation method thereof, wherein the calcium alginate liquid core microcapsule comprises the following components: the probiotic comprises one or more of lactobacillus, bifidobacterium, yeast, probiotic bacillus and clostridium butyricum. The calcium alginate liquid core microcapsule and the preparation method thereof provided by the invention can improve the freeze-drying survival rate and the storage survival rate of probiotics and the survival rate of gastrointestinal tracts by aiming at the problem that the survival rate of probiotics products is reduced in the processes of freeze drying, storage and gastrointestinal digestion.
Description
Technical Field
The invention relates to the technical field of probiotic microcapsules, in particular to a calcium alginate liquid core microcapsule and a preparation method thereof.
Background
Probiotics are defined as living microorganisms that have many health benefits to the host by ingestion of a certain amount. The presence of probiotics can enhance human immunity, relieve constipation, relieve lactose intolerance, resist gastrointestinal infection, etc. However, the human body must take a sufficient amount of probiotic bacteria orally and allow them to reach the small intestine and colonize the intestinal mucosa in order to exert the above-mentioned effects.
How to ensure the survival of probiotics in the whole process from production to final utilization by a human body is a critical problem which needs to be solved urgently. On one hand, probiotic products are often converted from a wet state to a dry state in order to facilitate transportation, storage and sale of the probiotic products, and most of the currently marketed drying processes for probiotic products are freeze drying. Freeze-drying has also proven to be an excellent way of drying probiotics which are heat sensitive. However, the formation of ice crystals during freezing and solute effects during drying inevitably have a detrimental effect on the probiotic bacteria, causing cell membrane damage, inactivation of active proteins and DNA damage to the probiotic bacteria, resulting in a decreased survival rate of the freeze-dried probiotic bacteria. On the other hand, probiotic products often undergo a storage period before they are consumed. During storage, the probiotics are influenced by the environment such as temperature, humidity and oxygen, and cell death is caused, so that the storage survival rate of the probiotics is reduced. In addition to this, during the course of the probiotic product through the human gastrointestinal tract, hydrogen ions in the gastric acid cause the zeta potential of the ingested probiotic to change, thereby altering the cell surface permeability and leading to the death of the probiotic. The existence of bile salts in the intestinal tract can also destroy the membrane structure of the probiotics, and the probiotics are oxidized and damaged to be inactivated.
In order to solve the above problem of probiotic survival rate, researchers have conducted studies using various methods. Selection of tolerant strains can be facilitated by in vitro screening, but the process is time consuming and complicated and difficult. Tolerance can be conferred to probiotics by genetic engineering, but the use of probiotic products in food products is limited. In contrast, microcapsule technology is an effective means. The development of microcapsules which are simple in preparation method, natural and safe, can be industrialized and can improve the survival of probiotics is urgently needed.
Disclosure of Invention
This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.
The present invention has been made in view of the above and/or other problems associated with the prior art calcium alginate liquid core microcapsule products.
Therefore, one of the purposes of the invention is to overcome the defects of the existing products and provide a calcium alginate liquid core microcapsule.
To solve the above technical problem, according to an aspect of the present invention, the present invention provides the following technical solutions: the calcium alginate liquid core microcapsule comprises the following components: the probiotic comprises one or more of lactobacillus, bifidobacterium, yeast, probiotic bacillus and clostridium butyricum.
The invention also aims to provide a preparation method of the calcium alginate liquid core microcapsule.
To solve the above technical problem, according to an aspect of the present invention, the present invention provides the following technical solutions: a method for preparing calcium alginate liquid core microcapsules comprises the following steps:
preparing a bacterial suspension: inoculating probiotics into a liquid culture medium for culture and carrying out centrifugal cleaning to obtain a bacterial suspension;
preparing core material liquid: weighing calcium ions, dissolving in water, stirring to dissolve completely, adding trehalose, stirring to dissolve completely, weighing appropriate amount of polysaccharide, and stirring to dissolve completely to obtain core material liquid;
adding a bacterial suspension: adding the bacterial suspension into the core material liquid and stirring uniformly;
preparing a coagulation bath liquid: weighing sodium alginate in deionized water, stirring until the sodium alginate is completely dissolved, standing and degassing;
preparing microcapsules: adding the prepared core material liquid into a coagulating bath, filtering after a period of time, and flushing to obtain microcapsules;
hardening: the microcapsules obtained are poured into a calcium ion solution for hardening, filtered and rinsed after a period of time.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: in the core material liquid preparation, the polysaccharide is thickening polysaccharide.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: in the core material liquid preparation, the calcium ions are calcium chloride or calcium carbonate or calcium lactate or calcium edetate, and in the hardening, the calcium ions are one or more of calcium chloride or calcium carbonate or calcium lactate or calcium edetate.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: in the core material liquid, the thickening polysaccharide comprises one or more of hyaluronic acid, xanthan gum, gellan gum, guar gum, locust bean gum, tamarind gum and sodium carboxymethylcellulose.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: in the core material liquid preparation, the concentration of the sodium alginate solution is 0.4-0.8% of the mass volume ratio.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: in the core preparation liquid, the concentration of calcium ions is 0.7-2.5% by mass volume ratio.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: in the core material liquid, the concentration of the trehalose is 1.5-5.5% by mass volume ratio.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: in the core material liquid preparation, the concentration of the thickening polysaccharide is 0.3-0.6% by mass volume ratio.
According to a preferable scheme of the preparation method of the calcium alginate liquid core microcapsule, the preparation method comprises the following steps: during hardening, the concentration of calcium ions in the calcium ion solution is 0.7-2.5% by mass volume ratio. The invention provides a calcium alginate liquid core microcapsule and a preparation method thereof, and provides a preparation method of the calcium alginate liquid core microcapsule for improving the survival rate of probiotics aiming at the problem of the survival rate reduction of probiotics products in the processes of freeze drying, storage and gastrointestinal digestion.
The invention uses sodium alginate as coagulating bath, and uses solution containing probiotics, calcium ions, trehalose and thickening polysaccharide as core material liquid. Wherein the calcium ions include calcium chloride, calcium carbonate, calcium lactate, calcium edetate, etc., and the thickening polysaccharide includes hyaluronic acid, xanthan gum, gellan gum, guar gum, locust bean gum, tamarind gum, sodium carboxymethylcellulose, etc.
The invention drops the core material into the coagulation bath in stirring by a simple extrusion method to spontaneously form the probiotic-loaded liquid core microcapsule with the calcium alginate biofilm, and the microcapsule is filtered and washed, then put into a calcium ion solution to be hardened, and finally filtered and washed.
Practice proves that the probiotics microcapsule prepared by the invention patent can improve the freeze drying survival rate and the storage survival rate of probiotics and can also improve the survival rate of gastrointestinal tracts.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof are described in detail below with reference to examples of the specification.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. The following examples are set forth to enable those skilled in the art to practice the invention.
Example 1
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Weighing sodium alginate in deionized water, wherein the concentration of the sodium alginate is 0.4-0.8% (w/v), stirring until the sodium alginate is completely dissolved, standing and degassing.
(4) Weighing calcium chloride in deionized water with the concentration of 0.7-2.5% (w/v), and stirring until the calcium chloride is completely dissolved.
(5) And (4) weighing trehalose in the solution in the step (4) with the concentration of 1.5-5.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing hyaluronic acid in the solution in the step (5) with the concentration of 0.3-0.6% (w/v), stirring until the hyaluronic acid is completely dissolved, and standing for degassing.
(7) And (3) adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring.
(8) Dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) And pouring the microcapsules into a newly prepared calcium chloride solution for hardening, wherein the concentration is 0.7-2.5% (w/v), the mass ratio of the microcapsules to the calcium chloride is 70:3, and filtering and washing the microcapsules by using a filter screen with the aperture of 500 mu m after 10 minutes.
Example 2
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with other apparatus material in autoclave at 115 deg.C for 20 min;
(2) inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension;
(3) weighing sodium alginate in deionized water, wherein the concentration of the sodium alginate is 0.4-0.8% (w/v), stirring until the sodium alginate is completely dissolved, standing and degassing;
(4) weighing calcium chloride in deionized water, wherein the concentration of the calcium chloride is 0.7-2.5% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) weighing trehalose in the solution obtained in the step (4), wherein the concentration of the trehalose is 1.5-5.5% (w/v), and stirring until the trehalose is completely dissolved;
(6) weighing xanthan gum in the solution obtained in the step (5) with the concentration of 0.3-0.6% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing;
(7) adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotating speed of 1000rpm by using a flow pump, continuously stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min;
(9) and pouring the microcapsules into a newly prepared calcium chloride solution for hardening, wherein the concentration is 0.7-2.5% (w/v), the mass ratio of the microcapsules to the calcium chloride is 70:3, and filtering and washing the microcapsules by using a filter screen with the aperture of 500 mu m after 10 minutes.
Example 3
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) and (3) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotating speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 4
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 0.7% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) and (3) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotating speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 5
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 2.5% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 6
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with a concentration of 1.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) and (3) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirring at the rotating speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 7
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 5.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) and (3) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotating speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 8
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.3% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 9
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.6% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 10
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate was weighed into deionized water at a concentration of 0.4% (w/v), stirred to dissolve completely and degassed by standing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 11
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.8 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) and (3) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirring at the rotating speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 12
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium carbonate was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) and (3) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirring at the rotating speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 13
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium lactate was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) and (3) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirring at the rotating speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 14
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium edetate was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing xanthan gum into the solution in the step (5) with the concentration of 0.45% (w/v), stirring until the xanthan gum is completely dissolved, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 15
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing hyaluronic acid in the solution in the step (5) to obtain a concentration of 0.45% (w/v), stirring until the hyaluronic acid is completely dissolved, and standing for degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 16
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing gellan gum into the solution in step (5) with a concentration of 0.45% (w/v), stirring until complete dissolution and standing for degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 17
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing locust bean gum in the solution of the step (5) with the concentration of 0.45% (w/v), stirring to completely dissolve, standing and degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 18
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing tamarind gum in the solution of step (5) at a concentration of 0.45% (w/v), stirring to dissolve completely, and standing for degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 19
(1) Preparing liquid culture medium, solid culture medium, 0.85% physiological saline, and sterilizing with autoclave at 115 deg.C for 20 min.
(2) Inoculating frozen Lactobacillus rhamnosus LGG into liquid culture medium, anaerobically culturing for 24 hr, centrifuging (4 deg.C, 3500g, 10 min), and cleaning for 2 times to obtain bacterial suspension.
(3) Sodium alginate is weighed into deionized water with the concentration of 0.6 percent (w/v), stirred until the sodium alginate is completely dissolved, and then kept stand for degassing.
(4) Calcium chloride was weighed into deionized water at a concentration of 1.6% (w/v) and stirred until completely dissolved.
(5) Weighing trehalose in the solution in the step (4) to obtain a solution with the concentration of 3.5% (w/v), and stirring until the trehalose is completely dissolved.
(6) Weighing sodium carboxymethylcellulose into the solution in the step (5) at a concentration of 0.45% (w/v), stirring until the sodium carboxymethylcellulose is completely dissolved, and standing for degassing.
(7) Adding the bacterial suspension obtained in the step (2) into the core material liquid obtained in the step (6) and fully stirring;
(8) dropwise adding the core material obtained in the step (7) into the coagulating bath obtained in the step (3) and stirred at the rotation speed of 1000rpm by using a flow pump, continuing stirring for 3min after the addition is finished, and filtering and washing by using a filter screen with the aperture of 500 mu m after the stirring is finished for 10 min.
(9) The microcapsules were poured into a freshly prepared calcium chloride solution for hardening at a concentration of 1.6% (w/v) in a mass ratio of microcapsules to calcium chloride of 70:3, and after 10 minutes filtered and rinsed using a filter mesh with a pore size of 500 μm.
Example 20
The freeze-drying survival rate, the storage survival rate, the gastric acid survival rate and the bile salt resistance survival rate of the calcium alginate liquid core microcapsule loaded with the probiotics prepared in the examples 1 to 19 are measured and recorded in the table 1, and the measuring method is as follows:
determining the freeze-drying survival rate: the resulting gel beads were frozen at-80 ℃ for 12 hours and then freeze-dried for 48 hours.
Determination of the storage survival rate: placing the lyophilized gel beads at 25 deg.C, controlling water activities at 0.113, 0.313, 0.501, and 0.812 respectively, and storing for 60 days.
Determining the gastric acid-resistant survival rate: the freeze-dried gel beads were placed in simulated gastric fluid (pH 2) in vitro for 2 hours.
Determining the survival rate of the bile salt: the freeze-dried gel beads were placed in a simulated bile salt solution (pH 6.8) for 1 hour.
TABLE 1 Probiotics-loaded calcium alginate liquid core microcapsules prepared in examples 1-19 Performance of probiotic stored probiotic
As can be seen from table 1, the microcapsules prepared in my invention have good loading effect on probiotics and good applicable effect on various polysaccharides, thickening polysaccharides and calcium ions, and the parameters of the preparation steps in the invention are preferably as shown in example 3.
It should be noted that the above-mentioned embodiments are only for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, which should be covered by the claims of the present invention.
Claims (10)
1. A method for preparing calcium alginate liquid core microcapsule is characterized in that: comprises the following components: the probiotic comprises one or more of lactobacillus, bifidobacterium, yeast, probiotic bacillus and clostridium butyricum.
2. A method for preparing calcium alginate liquid core microcapsule is characterized in that: the method comprises the following steps:
preparing a bacterial suspension: inoculating probiotics into a liquid culture medium for culture and carrying out centrifugal cleaning to obtain a bacterial suspension;
preparing core material liquid: weighing calcium ions, dissolving in water, stirring to dissolve completely, adding trehalose, stirring to dissolve completely, weighing appropriate amount of polysaccharide, and stirring to dissolve completely to obtain core material liquid;
adding a bacterial suspension: adding the bacterial suspension into the core material liquid and stirring uniformly;
preparing a coagulation bath liquid: weighing sodium alginate in deionized water, stirring until the sodium alginate is completely dissolved, standing and degassing;
preparing microcapsules: adding the prepared core material liquid into a coagulating bath, filtering after a period of time, and flushing to obtain microcapsules;
hardening: the microcapsules obtained are poured into a calcium ion solution for hardening, filtered and rinsed after a period of time.
3. The method for preparing calcium alginate liquid core microcapsule according to claim 2, characterized in that: in the core material preparation liquid, the polysaccharide is thickening polysaccharide.
4. The method for preparing calcium alginate liquid core microcapsule according to claim 2, characterized in that: in the core preparation liquid, calcium ions are calcium chloride or calcium carbonate or calcium lactate or calcium edetate, and in hardening, the calcium ions are one or more of calcium chloride or calcium carbonate or calcium lactate or calcium edetate.
5. The method for preparing calcium alginate liquid core microcapsule according to claim 3, characterized in that: in the core material preparation liquid, the thickening polysaccharide comprises one or more of hyaluronic acid, xanthan gum, gellan gum, guar gum, locust bean gum, tamarind gum and sodium carboxymethylcellulose.
6. The method for preparing calcium alginate liquid core microcapsule according to claim 2, characterized in that: in the core preparation liquid, the concentration of the sodium alginate solution is 0.4-0.8% of the mass volume ratio.
7. The method for preparing calcium alginate liquid core microcapsule according to claim 2, characterized in that: in the core preparation liquid, the concentration of calcium ions is 0.7-2.5% by mass volume ratio.
8. The method for preparing calcium alginate liquid core microcapsule according to claim 2, characterized in that: in the core preparation liquid, the concentration of trehalose is 1.5-5.5% by mass volume ratio.
9. The method for preparing calcium alginate liquid core microcapsule according to claim 5, characterized in that: in the core material liquid, the concentration of the thickening polysaccharide is 0.3-0.6% by mass volume ratio.
10. The method for preparing calcium alginate liquid core microcapsule according to claim 2, characterized in that: in the hardening, the concentration of calcium ions in the calcium ion solution is 0.7-2.5% by mass volume ratio.
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