CN117731009A - Infant probiotic supplementing chewing gel and preparation method thereof - Google Patents
Infant probiotic supplementing chewing gel and preparation method thereof Download PDFInfo
- Publication number
- CN117731009A CN117731009A CN202311674062.6A CN202311674062A CN117731009A CN 117731009 A CN117731009 A CN 117731009A CN 202311674062 A CN202311674062 A CN 202311674062A CN 117731009 A CN117731009 A CN 117731009A
- Authority
- CN
- China
- Prior art keywords
- gel
- probiotic
- chewing
- cross
- culture medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000006041 probiotic Substances 0.000 title claims abstract description 58
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 58
- 230000001055 chewing effect Effects 0.000 title claims abstract description 44
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title abstract description 20
- 230000001502 supplementing effect Effects 0.000 title abstract description 9
- 238000001879 gelation Methods 0.000 title description 2
- 150000004676 glycans Chemical class 0.000 claims abstract description 19
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 19
- 239000005017 polysaccharide Substances 0.000 claims abstract description 19
- 244000005700 microbiome Species 0.000 claims abstract description 9
- 230000003305 autocrine Effects 0.000 claims abstract description 6
- 239000010410 layer Substances 0.000 claims abstract description 5
- 230000009172 bursting Effects 0.000 claims abstract description 3
- 239000011241 protective layer Substances 0.000 claims abstract description 3
- 239000001963 growth medium Substances 0.000 claims description 69
- 238000009630 liquid culture Methods 0.000 claims description 58
- 239000000243 solution Substances 0.000 claims description 48
- 238000003756 stirring Methods 0.000 claims description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 36
- 238000005303 weighing Methods 0.000 claims description 36
- 230000001580 bacterial effect Effects 0.000 claims description 29
- 239000000725 suspension Substances 0.000 claims description 28
- 239000008367 deionised water Substances 0.000 claims description 24
- 229910021641 deionized water Inorganic materials 0.000 claims description 24
- 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 23
- 239000000661 sodium alginate Substances 0.000 claims description 23
- 235000010413 sodium alginate Nutrition 0.000 claims description 23
- 229940005550 sodium alginate Drugs 0.000 claims description 23
- 239000003431 cross linking reagent Substances 0.000 claims description 19
- 238000000855 fermentation Methods 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 13
- 238000004140 cleaning Methods 0.000 claims description 12
- 239000008223 sterile water Substances 0.000 claims description 12
- 238000004132 cross linking Methods 0.000 claims description 11
- 241000186000 Bifidobacterium Species 0.000 claims description 7
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 6
- 241000186660 Lactobacillus Species 0.000 claims description 3
- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 claims description 3
- 229910001626 barium chloride Inorganic materials 0.000 claims description 3
- 229940045511 barium chloride Drugs 0.000 claims description 3
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 3
- 229960003563 calcium carbonate Drugs 0.000 claims description 3
- 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 3
- 239000001527 calcium lactate Substances 0.000 claims description 3
- 229960002401 calcium lactate Drugs 0.000 claims description 3
- 235000011086 calcium lactate Nutrition 0.000 claims description 3
- 239000000679 carrageenan Substances 0.000 claims description 3
- 235000010418 carrageenan Nutrition 0.000 claims description 3
- 229920001525 carrageenan Polymers 0.000 claims description 3
- 229940113118 carrageenan Drugs 0.000 claims description 3
- 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 3
- 229940039696 lactobacillus Drugs 0.000 claims description 3
- 235000010987 pectin Nutrition 0.000 claims description 3
- 239000001814 pectin Substances 0.000 claims description 3
- 229960000292 pectin Drugs 0.000 claims description 3
- 229920001277 pectin Polymers 0.000 claims description 3
- 229910001631 strontium chloride Inorganic materials 0.000 claims description 3
- 229940013553 strontium chloride Drugs 0.000 claims description 3
- AHBGXTDRMVNFER-UHFFFAOYSA-L strontium dichloride Chemical compound [Cl-].[Cl-].[Sr+2] AHBGXTDRMVNFER-UHFFFAOYSA-L 0.000 claims description 3
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 claims description 3
- 230000004151 fermentation Effects 0.000 claims description 2
- 238000011010 flushing procedure Methods 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 230000000153 supplemental effect Effects 0.000 claims 7
- 230000004083 survival effect Effects 0.000 abstract description 9
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 7
- 230000029087 digestion Effects 0.000 abstract description 3
- 239000000499 gel Substances 0.000 description 70
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 40
- 239000001110 calcium chloride Substances 0.000 description 40
- 229910001628 calcium chloride Inorganic materials 0.000 description 40
- 239000007787 solid Substances 0.000 description 12
- 239000000463 material Substances 0.000 description 11
- 239000000203 mixture Substances 0.000 description 10
- 239000002504 physiological saline solution Substances 0.000 description 10
- 230000001954 sterilising effect Effects 0.000 description 10
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 8
- 239000013589 supplement Substances 0.000 description 7
- 238000012258 culturing Methods 0.000 description 6
- 210000004211 gastric acid Anatomy 0.000 description 6
- 210000000214 mouth Anatomy 0.000 description 5
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 239000004380 Cholic acid Substances 0.000 description 4
- 240000006024 Lactobacillus plantarum Species 0.000 description 4
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 4
- 235000019416 cholic acid Nutrition 0.000 description 4
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 4
- 229960002471 cholic acid Drugs 0.000 description 4
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 229940072205 lactobacillus plantarum Drugs 0.000 description 4
- 230000008901 benefit Effects 0.000 description 3
- 239000003833 bile salt Substances 0.000 description 3
- 241000186012 Bifidobacterium breve Species 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000018612 quorum sensing Effects 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 230000006355 external stress Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002633 protecting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a probiotic chewing gel for supplementing children and a preparation method thereof, which comprises a shell of polysaccharide gel and an inner layer of microorganism and microorganism autocrine protective layer tissues, wherein the inner layer is wrapped by the shell of polysaccharide gel, and the bursting strength of the inner layer is less than 900g. The invention can still keep higher gastrointestinal tract survival rate after chewing in the mouth, has the characteristics of lower chewing and biting force requirement and convenience in digestion, and can have good probiotic supplementing effect on infants.
Description
Technical Field
The invention belongs to the technical field of probiotic supplements, and particularly relates to a probiotic chewing gel for supplementing infants and a preparation method thereof.
Background
Probiotics are active microorganisms which have health effects on hosts after being ingested in certain amounts, and have various benefits on human bodies such as regulating intestinal flora, relaxing bowels, moistening stool, relieving inflammation and the like. The probiotics only reach the colon in sufficient quantity to exert the above effects. However, gastric acid and bile salts in the gastrointestinal tract are two major factors in stress probiotic survival, making the probiotics capable of reaching the colon in an active state severely deficient.
Embedding technology is an effective strategy to solve the above-mentioned problems. Entrapment is the reduction of probiotic inactivation in the gastrointestinal tract by enclosing the probiotic in a wall material, providing additional physical protection to the probiotic. The gel based on the biomacromolecule can effectively embed probiotics, and can realize the mild compatibility of the probiotics and the gel. The extensive research of the gel promotes the diversified development of the commodity type loaded with probiotics, and especially the probiotic jelly which is emerging in recent years is popular with consumers. However, while researchers have designed various gel types and structures to allow the loaded probiotics to exhibit better gastrointestinal tolerance, the necessary links to oral chewing that the gel presents when consumed by consumers are ignored. Chewing typically breaks the gel structure, re-exposing the probiotics to gastric acid and bile salt environments, and thereby losing the gel protecting effect. Therefore, how to combine the chewability of the gel with the high activity of the probiotics in the gastrointestinal tract is a critical issue to be solved by commercial products of gel-type probiotics.
In nature, microorganisms often form colonies that aggregate together to resist the harsh environment of the outside world. In this process, microorganisms develop quorum sensing effects by autocrine extracellular polymeric substances and transmitting intercellular molecular signals within the enclosed space, thereby achieving high resistance to external stress. In light of the above, we have resorted to gel structures to provide the necessary conditions for the formation of probiotic-mediated quorum sensing, thereby achieving the protective ability of the autocrine extracellular polymer to the bacterial cells. Thereby achieving a compromise between the chewability of the gel and the high activity of the probiotics in the gastrointestinal tract.
At present, the probiotic supplements are mostly carried out in the form of aqueous solution, the soft candy gel type supplements are carried out completely according to the chewing force of adults, and the probiotic supplements carried out by children are completely absent and cannot cope with the current problem of solving the probiotic supplements of infants
Disclosure of Invention
This section is intended to outline some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section as well as in the description summary and in the title of the application, to avoid obscuring the purpose of this section, the description summary and the title of the invention, which should not be used to limit the scope of the invention.
The present invention has been made in view of the above and/or problems occurring in the prior art.
Therefore, the invention aims to overcome the defects in the prior art and provide the probiotic-supplementing chewing gel for infants.
In order to solve the technical problems, the invention provides the following technical scheme: a baby supplement probiotic chewing gel comprising an outer shell of polysaccharide gel and an inner layer of microorganism and microorganism autocrine protective layer tissue encased therein, the baby supplement probiotic chewing gel having a burst strength of less than 900g.
The invention further aims at providing a preparation method of the infant probiotic-supplementing chewing gel.
In order to solve the technical problems, the invention provides the following technical scheme: a method for preparing a chewing gel for supplementing probiotics for infants comprises,
inoculating frozen probiotics into a liquid culture medium for anaerobic culture for 24 hours, centrifuging and cleaning for 2 times to prepare bacterial suspension;
weighing cross-linking polysaccharide in deionized water, and stirring until the cross-linking polysaccharide is completely dissolved to obtain a cross-linking polysaccharide solution;
weighing and dissolving a crosslinking agent in deionized water, and stirring until the crosslinking agent is completely dissolved to prepare a crosslinking agent solution;
weighing and dissolving the cross-linking agent in the liquid culture medium, and stirring until the cross-linking agent is completely dissolved to prepare a cross-linking agent-liquid culture medium;
adding the bacterial suspension into a cross-linked polysaccharide solution, stirring to obtain a mixed solution, and adding a cross-linking agent solution to form gel loaded with probiotics;
placing the gel loaded with the probiotics into a cross-linking agent-liquid culture medium, carrying out anaerobic fermentation on the gel in an anaerobic workstation, taking the gel, and flushing the gel with sterile water to obtain the infant probiotic-supplementing chewing gel.
As a preferred scheme of the preparation method of the infant probiotic chewing gel, the preparation method comprises the following steps: the prepared bacterial suspension is characterized in that the probiotics are one or more of lactobacillus and bifidobacterium.
As a preferred scheme of the preparation method of the infant probiotic chewing gel, the preparation method comprises the following steps: the crosslinkable polysaccharide comprises one or more of sodium alginate, carrageenan and pectin.
As a preferred scheme of the preparation method of the infant probiotic chewing gel, the preparation method comprises the following steps: the cross-linking agent comprises one or more of strontium chloride, barium chloride, calcium carbonate, calcium lactate and calcium edetate.
As a preferred scheme of the preparation method of the infant probiotic chewing gel, the preparation method comprises the following steps: the concentration of the crosslinkable polysaccharide in the crosslinkable polysaccharide solution is 0.3-5.0% w/v.
As a preferred scheme of the preparation method of the infant probiotic chewing gel, the preparation method comprises the following steps: the cross-linking agent solution, wherein the concentration of the cross-linking agent is 0.5-5.0% w/v.
As a preferred scheme of the preparation method of the infant probiotic chewing gel, the preparation method comprises the following steps: the concentration of the bacterial suspension is 10 3 ～10 8 CFU/mL。
As a preferred scheme of the preparation method of the infant probiotic chewing gel, the preparation method comprises the following steps: the anaerobic fermentation is carried out, wherein the fermentation time is 1.0-5.0 days.
The invention has the beneficial effects that:
the invention provides a probiotic chewing gel for supplementing children and a preparation method thereof, wherein the probiotic chewing gel is used and supplemented for probiotics including lactobacillus, bifidobacterium and the like, and the cross-linking polysaccharide comprises sodium alginate, carrageenan, pectin and the like; under the condition of using strontium chloride, barium chloride, calcium carbonate, calcium lactate, calcium edetate and the like, the cross-linking agent can still keep higher gastrointestinal survival rate after chewing the oral cavity, has the characteristics of lower chewing and biting force requirement and convenience in digestion, and can have good probiotic supplementing effect on infants.
Detailed Description
In order that the above-recited objects, features and advantages of the present invention will become more apparent, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
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 other than those described herein, and persons skilled in the art will readily appreciate that the present invention is not limited to the specific embodiments disclosed below.
Further, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic can be 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.
Example 1
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating lactobacillus rhamnosus LGG into MRS liquid culture medium, anaerobically culturing for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 0.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 5 CFU/mL) is added into the solution in the step (3) and stirred, wherein, the volume isThe ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at 1:20 (v/v) to form a lactobacillus rhamnosus LGG-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 2.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 2
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen lactobacillus rhamnosus LGG into MRS liquid culture medium, anaerobically culturing for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 0.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 7 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at 1:20 (v/v) to form a lactobacillus rhamnosus LGG-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 2.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 3
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen lactobacillus rhamnosus LRa05 into MRS liquid culture medium, anaerobically culturing for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 0.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 5 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at 1:20 (v/v) to form a gel loaded with lactobacillus rhamnosus LRa 05;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 1.0 day;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 4
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen lactobacillus rhamnosus LRa05 into MRS liquid culture medium, anaerobically culturing for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 0.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 7 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at 1:20 (v/v) to form a gel loaded with lactobacillus rhamnosus LRa 05;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 1.0 day;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 5
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen lactobacillus plantarum Lp05 into MRS liquid culture medium, performing anaerobic culture for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 1.0% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 2.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 2.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 4 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at a ratio of 1:20 (v/v) to form a lactobacillus plantarum Lp 05-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 3.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 6
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen lactobacillus plantarum Lp05 into MRS liquid culture medium, performing anaerobic culture for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 1.0% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 6 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at a ratio of 1:20 (v/v) to form a lactobacillus plantarum Lp 05-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 2.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 7
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen bifidobacterium BB12 into MRS liquid culture medium, performing anaerobic culture for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 1.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 2.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 2.5% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 6 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at a ratio of 1:20 (v/v) to form a bifidobacterium BB 12-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 4.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 8
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen bifidobacterium BB12 into MRS liquid culture medium, performing anaerobic culture for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 0.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 8 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at a ratio of 1:20 (v/v) to form a bifidobacterium BB 12-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 3.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 9
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen Bradyyeast into MRS liquid culture medium, anaerobic culturing for 24 hr, centrifuging (4deg.C, 5000g,10 min), and cleaning for 2 times to obtain bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 1.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 2.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 2.5% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 6 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at 1:20 (v/v) to form a buddyyeast-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 2.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 10
(1) Preparing a liquid culture medium, a solid culture medium and 0.85% physiological saline, and sterilizing the mixture for 20 minutes at 115 ℃ by using an autoclave together with other appliance materials;
(2) Inoculating frozen bifidobacterium breve BBr60 into MRS liquid culture medium, anaerobic culturing for 24 hours, centrifuging (4 ℃ C., 5000g,10 minutes), and cleaning for 2 times to prepare bacterial suspension;
(3) Weighing sodium alginate in deionized water to a concentration of 0.5% (w/v), and stirring until the sodium alginate is completely dissolved;
(4) Weighing calcium chloride in deionized water to a concentration of 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(5) Weighing calcium chloride in a liquid culture medium, wherein the concentration is 1.0% (w/v), and stirring until the calcium chloride is completely dissolved;
(6) Taking the bacterial suspension (10) of the step (2) 7 CFU/mL) is added into the solution in the step (3) and stirred, wherein the volume ratio is 1:20;
(7) Adding the solution of step (6) to the solution of step (4) at a ratio of 1:20 (v/v) to form a bifidobacterium breve BBr 60-loaded gel;
(8) Placing the gel obtained in the step (7) in the liquid culture medium in the step (5) at a ratio of 1:50 (w/w);
(9) Placing the liquid culture medium in the step (8) in an anaerobic workstation (37 ℃) for anaerobic fermentation for 2.0 days;
(10) The gel in step (9) was removed and rinsed with sterile water.
Example 11
The probiotic-loaded gels prepared in examples 1 to 10 were subjected to simulated chewing in the mouth, followed by gastric acid and cholic acid resistant survival measurements recorded in table 1, the measurements being as follows:
method for determining gastric acid-resistant survival rate: the gel after 0.5g of simulated chewing of the oral cavity (gel beads are broken for 5S by a nine-yang 8S speed stirring gear stirrer) is placed in vitro simulated gastric juice (pH=2) stirred at 600rpm for 2 hours, then placed in 5ml of 0.05M sodium citrate solution for vortex, then subjected to gradient dilution, mixed with solid culture medium by a pouring method, anaerobic cultured at 37 ℃ for 48 hours, and counted.
Gastric acid resistant survival = number of post gastric acid bacteria (CFU/g)/number of post freeze-drying bacteria (CFU/g) ×100%
Method for determining survival rate of cholic acid:
0.5g of gel after simulated chewing of the oral cavity was placed in an in vitro simulated bile salt solution (ph=6.8) stirred at 600rpm for 1 hour, then placed in 5ml of 0.05M sodium citrate solution, vortexed to release capsules, then diluted in gradient, poured into a mixed solid medium, anaerobic cultured at 37 ℃ for 48 hours, and counted.
Cholic acid survival = number of post cholic acid bacteria (CFU/g)/number of post freeze-drying bacteria (CFU/g) ×100%
Table 1 viability data and burst force of gel beads prepared in examples 1 to 10
In Table 1, example 9 did not give good results, probably because the strain was insufficient in the ability to autocrine extracellular polymer under this condition. The gel finished product prepared in the invention has obvious gastrointestinal tract tolerance to most probiotics after chewing, has a small advantage in bursting force, and is convenient to adapt to the chewing scene of infants.
The invention can still keep higher gastrointestinal tract survival rate after chewing in the mouth, has the characteristics of lower chewing and biting force requirement and convenience in digestion, and can have good probiotic supplementing effect on infants.
It should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, 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 the technical solution of the present invention may be modified or substituted without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered in the scope of the claims of the present invention.
Claims (9)
1. A probiotic-supplemented chewing gel for infants, characterized in that: an inner layer comprising an outer shell of polysaccharide gel, microorganisms entrapped therein, and a microorganism autocrine protective layer tissue;
the bursting strength of the infant probiotic-supplementing chewing gel is less than 900g.
2. A method of preparing a baby supplemental probiotic chewing gel according to claim 1, wherein: comprising the steps of (a) a step of,
inoculating frozen probiotics into a liquid culture medium for anaerobic culture for 24 hours, centrifuging and cleaning for 2 times to prepare bacterial suspension;
weighing cross-linking polysaccharide in deionized water, and stirring until the cross-linking polysaccharide is completely dissolved to obtain a cross-linking polysaccharide solution;
weighing and dissolving a crosslinking agent in deionized water, and stirring until the crosslinking agent is completely dissolved to prepare a crosslinking agent solution;
weighing and dissolving the cross-linking agent in the liquid culture medium, and stirring until the cross-linking agent is completely dissolved to prepare a cross-linking agent-liquid culture medium;
adding the bacterial suspension into a cross-linked polysaccharide solution, stirring to obtain a mixed solution, and adding a cross-linking agent solution to form gel loaded with probiotics;
placing the gel loaded with the probiotics into a cross-linking agent-liquid culture medium, carrying out anaerobic fermentation on the gel in an anaerobic workstation, taking the gel, and flushing the gel with sterile water to obtain the infant probiotic-supplementing chewing gel.
3. A method of preparing a baby supplemental probiotic chewing gel according to claim 2, wherein: the prepared bacterial suspension is characterized in that the probiotics are one or more of lactobacillus and bifidobacterium.
4. A method of preparing a baby supplemental probiotic chewing gel according to claim 2, wherein: the crosslinkable polysaccharide comprises one or more of sodium alginate, carrageenan and pectin.
5. A method of preparing a baby supplemental probiotic chewing gel according to claim 2, wherein: the cross-linking agent comprises one or more of strontium chloride, barium chloride, calcium carbonate, calcium lactate and calcium edetate.
6. A method of preparing a baby supplemental probiotic chewing gel according to claim 2 or 4, wherein: the concentration of the crosslinkable polysaccharide in the crosslinkable polysaccharide solution is 0.3-5.0% w/v.
7. A method of preparing a baby supplemental probiotic chewing gel according to claim 2 or 5, wherein: the cross-linking agent solution, wherein the concentration of the cross-linking agent is 0.5-5.0% w/v.
8. A method of preparing a baby probiotic-supplemented chewing gel according to claim 2 or 3, wherein: the concentration of the bacterial suspension is 10 3 ～10 8 CFU/mL。
9. A method of preparing a baby supplemental probiotic chewing gel according to claim 2 or 5, wherein: the anaerobic fermentation is carried out, wherein the fermentation time is 1.0-5.0 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311674062.6A CN117731009A (en) | 2023-12-07 | 2023-12-07 | Infant probiotic supplementing chewing gel and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311674062.6A CN117731009A (en) | 2023-12-07 | 2023-12-07 | Infant probiotic supplementing chewing gel and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117731009A true CN117731009A (en) | 2024-03-22 |
Family
ID=90258496
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311674062.6A Pending CN117731009A (en) | 2023-12-07 | 2023-12-07 | Infant probiotic supplementing chewing gel and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117731009A (en) |
-
2023
- 2023-12-07 CN CN202311674062.6A patent/CN117731009A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Silva et al. | Symbiotic microencapsulation to enhance Lactobacillus acidophilus survival | |
| Islam et al. | Microencapsulation of live probiotic bacteria | |
| CN109619593B (en) | Probiotic double-layer microcapsule and preparation method thereof | |
| Sathyabama et al. | Co-encapsulation of probiotics with prebiotics on alginate matrix and its effect on viability in simulated gastric environment | |
| CN108853021B (en) | Probiotic liquid preparation based on double-emulsion structure and preparation method thereof | |
| Prakash et al. | Artificial cell therapy: New strategies for the therapeutic delivery of live bacteria | |
| CN111588038A (en) | Seaweed enzyme probiotic microcapsule and preparation method thereof | |
| CN112956698B (en) | Explosion bead embedded with probiotics microcapsule and preparation method thereof | |
| JP4626752B2 (en) | Novel uses of carbohydrates and compositions | |
| CN114287632A (en) | Preparation method of inulin probiotic microcapsules | |
| WO2008111832A1 (en) | Method of improving skills with a composition comprising non-digestible saccharide | |
| CN116076731B (en) | Probiotic preparation with high immunity activity and preparation method thereof | |
| TWI425092B (en) | Bacterial cellulose composite with capsules embedded therein and preparation thereof | |
| CN113230280B (en) | Colon targeted probiotic multilayer embedded microcapsule and preparation method and application thereof | |
| CN110367542B (en) | Probiotic microcapsule slowly released in intestinal tract and preparation method thereof | |
| Shukla et al. | Pectin-based colon-specific drug delivery | |
| CN112335884A (en) | Novel probiotic microsphere and preparation method thereof | |
| CN113826903A (en) | Gastrointestinal tract controlled-release probiotic beads | |
| CN114916675A (en) | Water-in-oil-in-water type multiple emulsion gel bead for improving survival rate of probiotics, preparation method and application | |
| CN115944665A (en) | Probiotic agent for improving intestinal flora balance and preparation method and application thereof | |
| Oguntoye et al. | Viability of Lactobacillus rhamnosus GG in provitamin A cassava hydrolysate during fermentation, storage, in vitro and in vivo gastrointestinal conditions | |
| Lee et al. | Effect of pectic oligosaccharide on probiotic survival and physicochemical properties of hydrogel beads for synbiotic encapsulation of Lactobacillus bulgaricus | |
| Lai et al. | Microencapsulation of Bifidobacterium lactis Bi-07 with galactooligosaccharides using co-extrusion technique | |
| WO2020107580A1 (en) | Probiotic microcapsule with arabinoxylan-sodium alginate as wall material and preparation method therefor | |
| CN117731009A (en) | Infant probiotic supplementing chewing gel and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |