PREPARATION METHOD FOR DIET-REDUCING CAPSULE CONTAINING MULTI-NUTRIENT MICROSPHERES AND RESULTING PRODUCT
20240268439 ยท 2024-08-15
Assignee
Inventors
Cpc classification
A23P20/105
HUMAN NECESSITIES
A23P10/30
HUMAN NECESSITIES
International classification
A23P20/10
HUMAN NECESSITIES
A23L33/135
HUMAN NECESSITIES
Abstract
A preparation method for a diet-reducing capsule containing multi-nutrient microspheres and a resulting product, relating to the field of food biotechnology, are provided. The diet-reducing capsule includes a porous capsule shell, multi-nutrient microspheres and a matrix gel containing the multi-nutrient microspheres. The porous capsule shell is prepared by using a laser punching technology. The multi-nutrient microsphere is a miniature container which is formed by taking probiotics as a core material and taking a freeze-drying protective agent, a natural polymer material embedded with vitamins, minerals and prebiotics and an enteric coating material as a wall material. The matrix gel for the microspheres is biomimetic cellulose superabsorbent gel.
Claims
1. A preparation method for a diet-reducing capsule containing multi-nutrient microspheres, comprising the following steps: step (1), adding sodium carboxymethyl cellulose to an aqueous solution containing a polybasic carboxylic acid, stirring evenly to obtain a gel, drying the gel in a drying oven, then performing cross-linking at high temperature to obtain a cross-linked product, crushing and sieving the cross-linked product to obtain solid hydrogel particles, and washing and filtering with distilled water to prepare wet hydrogel particles; step (2), inoculating probiotics in a sterile medium, repeatedly activating the probiotics for 5 generations under the same culture conditions, collecting bacterial sludge by low-temperature centrifugation, washing the bacterial sludge with sterile normal saline, mixing the washed bacterial sludge evenly with an aqueous solution of a freeze-drying protective agent to obtain a bacterial suspension, then adding an aqueous solution of a natural polymer material to the bacterial suspension and mixing evenly again, then solidifying the obtained mixed solution, and washing and filtering to obtain solidified probiotics, followed by low-temperature storage for later use; step (3), mixing powders of vitamins, minerals and prebiotics evenly, sieving through a 70-mesh sieve to obtain a vitamin-mineral-prebiotics composite powder, and then mixing the vitamin-mineral-prebiotics composite powder evenly with the aqueous solution of the natural polymer material to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution; step (4), mixing the solidified probiotics evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution to obtain a mixed nutrient solution, then mixing the mixed nutrient solution with an aqueous solution of an enteric coating material to form microspheres, and washing and filtering the microspheres to obtain wet probiotics-prebiotics-vitamin-mineral microspheres; step (5), mixing the wet probiotics-prebiotics-vitamin-mineral microspheres evenly with the wet hydrogel particles, pre-cooling and then freeze-drying, and then crushing and sieving to obtain a capsule content; step (6), punching holes in an empty capsule shell with laser to obtain a porous capsule shell, and then combining the porous capsule shell with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres, wherein in the step (1), a viscosity of the sodium carboxymethyl cellulose is 7000 to 15000; a mass ratio of the sodium carboxymethyl cellulose to the polybasic carboxylic acid is (310 to 350):1, and a mass ratio of the sodium carboxymethyl cellulose to water is 1:(10 to 22); a way of stirring is performed first for 80 to 100 min at a rotation speed of 50 to 70 rpm, and then for 14 to 20 h at a rotation speed of 20 to 40 rpm; a drying temperature of the drying oven is 40 to 60? C., with drying first for 20 to 28 h, turning over and then continuing to dry for 28 to 36 h; a temperature of cross-linking at the high-temperature is 110 to 130? C., and a time of cross-linking at the high-temperature is 3.6 to 4.4 h; a way of crushing and sieving is to crush the cross-linked product with a crusher, and then sieve with an 18-mesh sieve and a 26-mesh sieve; a way of washing with the distilled water is to wash the solid hydrogel particles with the distilled water for 2 to 6 times, 2 to 4 h each time, wherein a mass ratio of the solid hydrogel particles to the distilled water is 1:(100 to 200) in each washing; in the step (6), a laser source of the laser is a cold light source, each punched hole has an aperture of 0.5 to 1.5 mm, and a number of the punched holes is 1 to 4; and when the porous capsule shell is combined with the capsule content, a mass of the capsule content contained in each porous capsule shell is 0.60 to 0.75 g.
2. The preparation method according to claim 1, wherein in the step (1), the polybasic carboxylic acid is one of citric acid, aconitic acid, oxalic acid, tartaric acid, malic acid, acetic acid, malonic acid, succinic acid, adipic acid, azelaic acid, terephthalic acid, trimellitic acid, trimesic acid, ethylenediaminetetraacetic acid, and 2-methylglutaric acid; in the step (2), the probiotics is a mixed strain selected from two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus salivarius, Lactobacillus helveticus, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus crispatus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophilus, Lactobacillus casei subsp. Casei, Lactobacillus paracasei, Lactobacillus reuteri, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium adolescent, Bifidobacterium animalis, and Streptococcus thermophilus; in the step (3), the vitamin is at least one of vitamin A, vitamin D3, vitamin E, vitamin K.sub.2, vitamin B.sub.1, vitamin B.sub.2, vitamin B.sub.6, vitamin B.sub.12, vitamin B.sub.13, vitamin B.sub.15, vitamin C, biotin, niacinamide, folic acid, inositol, and pantothenic acid; the mineral is at least one of calcium, magnesium, manganese, iron, zinc, cobalt, molybdenum, chromium, copper, selenium, iodine, phosphorus, potassium, sodium, sulfur, and chlorine; the prebiotics is one of fructooligosaccharides, xylooligosaccharides, galactooligosaccharides, isomaltooligosaccharides, soybean oligosaccharides, mannose oligosaccharides, lactulose, raffinose, stachyose, chitosan oligosaccharides, resistant starch, wheat dextrin, inulin, polydextrose, trehalose, Aspergillus niger oligosaccharides, spirulina, Arthrospira, chlorella, and microalgae; in the steps (2) and (3), the natural polymer material is one or more of sodium alginate, chitosan, modified starch, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, gallant gum, k-carrageenan, Arabic gum, pectin, carrageenan, gellan gum, xanthan gum, maltodextrin, ?-cyclodextrin, gelatin, soy protein isolate, and whey protein; in the step (2), the freeze-drying protective agent is one or more of soluble starch, hydroxyethyl starch, resistant dextrin, fructose, glucose, lactose, sucrose, ribose, rhamnose, galactose, fucose, mannose, arabinose, xylan, skimmed milk powder, glycerin, lactitol, sorbitol, mannitol, xylitol, erythritol, maltitol, sodium glutamate, antifreeze peptide, sericin peptide, fish collagen peptide, collagen, and polyvinylpyrrolidone; and in the step (4), the enteric coating material is one or more of shellac, algin, diclofenac, acrylic resin No. I, acrylic resin No. II, acrylic resin No. III, cellulose acetate benzenedicarboxylate, cellulose acetate succinate, hydroxypropyl methylcellulose acetate succinate, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, cellulose acetate 1,2,4-benzenetricarboxylate, hydroxypropyl methylcellulose 1,2,4-benzenetricarboxylate, hydroxypropyl methylcellulose titanate, and polyvinyl acetate phthalate.
3. The preparation method according to claim 1, wherein in the step (2), an inoculation amount of the probiotics is 1.5 to 4.5%; the sterile medium is an MRS liquid medium; the low-temperature centrifugation is to carry out centrifugation at 3 to 5? C., with a centrifugation speed of 3500 to 5500 rpm, and centrifugation time of 10 to 20 min; a number of times of washing with the sterile normal saline is 1 to 3, and a mass concentration of the sterile normal saline is 0.85% to 0.95%; in the aqueous solution of the freeze-drying protective agent, a mass fraction of the freeze-drying protective agent is 6% to 20%; the bacterial sludge and the aqueous solution of the freeze-drying protective agent are mixed in a volume ratio of 1:(3 to 5), and then stirred for 10 to 20 min at a stirring speed of 200 to 400 rpm; the probiotics in the bacterial suspension has a concentration of 109 CFU/mL; the probiotics is solidified with a 0.1 mol/L CaCl.sub.2) solution for 20 to 40 min; and a temperature of the low-temperature storage is 3 to 5?C.
4. The preparation method according to claim 1, wherein in the step (3), a ratio of a total mass of the vitamins, the minerals and the prebiotics to a mass of the aqueous solution of the natural polymer material is (1 to 2):11; the vitamin-mineral-prebiotics composite powder and the aqueous solution of the natural polymer material are mixed for 10 to 30 min at a rotation speed of 150 to 350 rpm; and in the vitamin-mineral-prebiotics-natural polymer material mixed solution, the vitamins comprise 95 to 128 ?g/g of vitamin A, 1 to 6 ?g/g of vitamin D3, 1 to 6 mg/g of vitamin E, 6 to 10 ?g/g of vitamin K.sub.2, 0.1 to 0.6 mg/g of vitamin B.sub.1, 0.1 to 0.6 mg/g of vitamin B.sub.2, 0.1 to 0.6 mg/g of vitamin B.sub.6, 0.1 to 0.7 ?g/g of vitamin B.sub.12, 1 to 7 mg/g of niacinamide, 40 to 80 ?g/g of folic acid, 10 to 40 mg/g of vitamin C, and 0.5 to 2.5 mg/g of pantothenic acid; the minerals comprise 93 to 133 mg/g of calcium carbonate, 27 to 51 mg/g of magnesium gluconate, 0.58 to 0.98 mg/g of manganese sulfate, 1 to 5 mg/g of ferrous lactate, 0.1 to 2.5 mg/g of zinc gluconate, 10 to 17 ?g/g of sodium selenite, and 0.01 to 0.30 mg/g of copper sulfate; and the prebiotics content is 0.3 to 1.0 g/100 mL.
5. The preparation method according to claim 1, wherein in the step (4), a mixing time of the solidified probiotics and the vitamin-mineral-prebiotics-natural polymer material mixed solution, and a mixing time of the mixed nutrient solution and the aqueous solution of the enteric coating material are 10 to 30 min, respectively, with a rotation speed of 100 to 300 rpm; a mass concentration of the aqueous solution of the enteric coating material is 4% to 12%; and the washing is performed with sterile distilled water for 2 to 4 times.
6. The preparation method according to claim 1, wherein in the step (5), a mixing time of the wet probiotics-prebiotics-vitamin-mineral microspheres and the wet hydrogel particles is 10 to 20 min, with a rotation speed of 30 to 60 rpm; a treatment of pre-cooling is performed at ?80? C. for 1 to 4 h; and conditions of freeze-drying are that: a temperature is ?55? C., a vacuum degree is 25 Pa, and a time is 24 to 48 h.
7. The preparation method according to claim 1, wherein in the step (6), the empty capsule shell is made of one of gelatin, pullulan and glutinous rice starch, and has a model selected from one of 000 #, 00 #, 0 #, 1 #, 2 #, 3 #, 4 # and extended models thereof; and a punching position of the empty capsule shell is one of symmetrical positions in the middle of the waist, central symmetrical positions at both ends, equidistant symmetrical positions at both ends and the waist, as well as equidistant positions of skew symmetry from the vertex and midline.
8. A diet-reducing capsule containing multi-nutrient microspheres which is prepared by the preparation method for the diet-reducing capsule containing the multi-nutrient microspheres according to claim 1.
9. The diet-reducing capsule containing the multi-nutrient microspheres according to claim 8, wherein the diet-reducing capsule comprises the porous capsule shell and the capsule content, and the capsule content is the microspheres containing multiple nutrients, and the solid hydrogel particles; the solid hydrogel particles are of a three-dimensional network structure composed of a polybasic carboxylic acid cross-linked with sodium carboxymethyl cellulose; the microsphere containing the multiple nutrients is a miniature container composed of the probiotics as a core material and protective layers as a wall material; the protective layers are a freeze-drying protective agent layer, a natural polymer material layer embedded with the vitamins, the minerals and the prebiotics, and an enteric coating material layer respectively from inside to outside; and the porous capsule shell is made by punching the holes in the capsule shell using the laser.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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[0075] In
##STR00001##
indicates that there is a hole in the front of the empty capsule shell,
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indicates that there is a hole in the back of the empty capsule shell, and
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indicates that there is a hole in the side of the empty capsule shell.
DESCRIPTION OF THE EMBODIMENTS
[0076] The present disclosure will be further described in detail below in conjunction with specific examples, comparative examples and accompanying drawings.
[0077] In the following examples and comparative examples, unless otherwise specified, the concentrations are all in percentage by mass.
[0078] In the following examples and comparative examples, unless otherwise specified, all sterilization operations in a sterile aqueous solution, an MRS medium, etc. are carried out by a moist heat sterilization method, wherein a sterilization temperature is 121? C. and the sterilization time is 20 min.
[0079] In the following examples and comparative examples, unless otherwise specified, in step (2), the probiotics is all mixed flora composed of Bifidobacterium longum and Lactobacillus acidophilus; the freeze-drying protective agents are soluble starch, skimmed milk powder, glycerol and xylan (a ratio of soluble starch to skimmed milk powder to glycerol to xylan is 5:6:1:10 in parts by weight), and a concentration of the probiotics in the bacterial suspension is 109 CFU/mL; in steps (2) and (3), the natural polymer materials are sodium alginate, chitosan and gellan gum (a ratio of sodium alginate to chitosan to gellan gum is 40:3:5 in parts by weight); in step (3), the prebiotics is fructooligosaccharides; in step (4), the enteric coating material is hydroxypropyl methylcellulose phthalate; and in step (6), all the empty capsule shells are made of gelatin, all in 00 # type, and a mass of the capsule content contained in the diet-reducing capsule is 0.75 g/capsule.
Example 1
[0080] Step (1), sodium carboxymethyl cellulose (a viscosity of 11000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 330:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:16) to obtain a mixture; the mixture was stirred at 60 rpm for 90 min, and then stirred for at 30 rpm 16 h to obtain a gel; the gel was dried in a drying oven at 45? C. for 24 h, turned over and continued to dry for 32 h, then cross-linked at high temperature of 120? C. for 4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 4 times, 3 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:150 in each washing), and filtered to obtain wet hydrogel particles;
[0081] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 3%, and repeatedly activated for 5 generations under the same culture conditions (36.5? C., 24 h); bacterial sludge was collected by low-temperature centrifugation (4? C., 4500 rpm, 15 min), and washed twice with 0.9% sterile normal saline; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 13% according to a volume ratio of 1:4 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1% according to a volume ratio of 1:1 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 30 min, washed and filtered to obtain solidified probiotics; the solidified probiotics was stored at a low temperature of 4?C for later use;
[0082] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (250 rpm, 20 min) evenly with the aqueous solution of the natural polymer material having a concentration of 3% according to a mass ratio of 1.5:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 111 ?g/g of vitamin A, 4 ?g/g of vitamin D3, 4 mg/g of vitamin E, 8 ?g/g of vitamin K.sub.2, 0.4 mg/g of vitamin B.sub.1, 0.4 mg/g of vitamin B.sub.2, 0.4 mg/g of vitamin B.sub.6, 0.4 ?g/g of vitamin B.sub.12, 4 mg/g of niacinamide, 60 ?g/g of folic acid, 25 mg/g of vitamin C, and 1.5 mg/g of pantothenic acid; 113 mg/g of calcium carbonate, 39 mg/g of magnesium gluconate, 0.78 mg/g of manganese sulfate, 3 mg/g of ferrous lactate, 1.5 mg/g of zinc gluconate, 14 ?g/g of sodium selenite, and 0.16 mg/g of copper sulfate; and the prebiotics content was 0.65 g/100 mL;
[0083] Step (4), the solidified probiotics was mixed (200 rpm, 20 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:3 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:5, washed for 3 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0084] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (45 rpm, 15 min) evenly with the wet hydrogel particles according to a mass ratio of 1:15 to obtain a mixture; the mixture was then pre-cooled at ?80? C. for 2.5 h, and then freeze-dried at ?55? ? C. for 36 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0085] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 1 mm, in symmetrical positions in the middle of the waist with laser (as shown in
Example 2
[0086] Step (1), sodium carboxymethyl cellulose (a viscosity of 15000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 350:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:10) to obtain a mixture; the mixture was first stirred at 70 rpm for 100 min, and then stirred at 40 rpm for 20 h to obtain a gel; the gel was dried in a drying oven at 60? C. for 28 h, turned over and continued to dry for 36 h, then cross-linked at high temperature of 125? C. for 4.4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 6 times, 4 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:200 in each washing), and filtered to obtain wet hydrogel particles;
[0087] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 4.5%, and repeatedly activated for 5 generations under the same culture conditions (38? C., 27 h); bacterial sludge was collected by low-temperature centrifugation (5? C., 5500 rpm, 20 min), and washed with 0.95% sterile normal saline for 3 times; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 20% according to a volume ratio of 1:5 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1.5% according to a volume ratio of 1:1.5 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 40 min, washed and filtered to obtain solidified probiotics; the solidified probiotics was stored at a low temperature of 5? ? C. for later use;
[0088] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (350 rpm, 30 min) evenly with the aqueous solution of the natural polymer material having a concentration of 4% according to a mass ratio of 2:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 128 ?g/g of vitamin A, 6 ?g/g of vitamin D3, 6 mg/g of vitamin E, 10 ?g/g of vitamin K.sub.2, 0.6 mg/g of vitamin B.sub.1, 0.6 mg/g of vitamin B.sub.2, 0.6 mg/g of vitamin B.sub.6, 0.7 ?g/g of vitamin B.sub.12, 7 mg/g of niacinamide, 80 ?g/g of folic acid, 40 mg/g of vitamin C, and 2.5 mg/g of pantothenic acid; 133 mg/g of calcium carbonate, 51 mg/g of magnesium gluconate, 0.98 mg/g of manganese sulfate, 5 mg/g of ferrous lactate, 2.5 mg/g of zinc gluconate, 17 ?g/g of sodium selenite, and 0.3 mg/g of copper sulfate; and the prebiotics content was 1.0 g/100 mL;
[0089] Step (4), the solidified probiotics was mixed (300 rpm, 30 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:4 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed evenly with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:6, washed for 4 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0090] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (60 rpm, 20 min) evenly with the wet hydrogel particles according to a mass ratio of 1:17 to obtain a mixture; the mixture was then pre-cooled at ?80? ? C. for 4 h, and then freeze-dried at ?55? C. for 48 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0091] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 1 mm, in symmetrical positions in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Example 3
[0092] Step (1), sodium carboxymethyl cellulose (a viscosity of 7000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 310:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:22) to obtain a mixture; the mixture was first stirred at 50 rpm for 80 min, and then stirred at 20 rpm for 14 h to obtain a gel; the gel was dried in a drying oven at 40? C. for 20 h, turned over and continued to dry for 28 h, then cross-linked at high temperature of 110? C. for 3.6 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed twice with distilled water, 2 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:100 in each washing), and filtered to obtain wet hydrogel particles;
[0093] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 1.5%, and repeatedly activated for 5 generations under the same culture conditions (35? C., 21 h); bacterial sludge was collected by low-temperature centrifugation (3? C., 3500 rpm, 10 min), and washed once with 0.85% sterile normal saline; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 6% according to a volume ratio of 1:3 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 0.5% according to a volume ratio of 1:0.5 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 20 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 3? C. for later use;
[0094] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (150 rpm, 10 min) evenly with the aqueous solution of the natural polymer material having a concentration of 3% according to a mass ratio of 1:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 95 ?g/g of vitamin A, 1 ?g/g of vitamin D3, 1 mg/g of vitamin E, 6 ?g/g of vitamin K.sub.2, 0.1 mg/g of vitamin B.sub.1, 0.1 mg/g of vitamin B.sub.2, 0.1 mg/g of vitamin B.sub.6, 0.1 ?g/g of vitamin B.sub.12, 1 mg/g of niacinamide, 40 ?g/g of folic acid, 10 mg/g of vitamin C, and 0.5 mg/g of pantothenic acid; the minerals include 93 mg/g of calcium carbonate, 27 mg/g of magnesium gluconate, 0.58 mg/g of manganese sulfate, 1 mg/g of ferrous lactate, 0.1 mg/g of zinc gluconate, 10 ?g/g of sodium selenite, and 0.01 mg/g of copper sulfate; and the prebiotics content was 0.3 g/100 mL;
[0095] Step (4), the solidified probiotics was mixed (100 rpm, 10 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:2 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed evenly with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:4, washed twice and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0096] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (30 rpm, 10 min) evenly with the wet hydrogel particles according to a mass ratio of 1:13 to obtain a mixture; the mixture was then pre-cooled at ?80? C. for 1 h, and then freeze-dried at ?55? C. for 24 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0097] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 1 mm, in central symmetrical positions at both ends of the entire shell with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Example 4
[0098] Step (1), sodium carboxymethyl cellulose (a viscosity of 11000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 350:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:10) to obtain a mixture; the mixture was first stirred at 60 rpm for 90 min, and then stirred at 40 rpm for 20 h to obtain a gel; the gel was dried in a drying oven at 40? ? C. for 20 h, turned over and continued to dry for 28 h, then cross-linked at high temperature of 120? ? C. for 4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 6 times, 4 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:200 in each washing), and filtered to obtain wet hydrogel particles;
[0099] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 1.5%, and repeatedly activated for 5 generations under the same culture conditions (36.5? C., 24 h); bacterial sludge was collected by low-temperature centrifugation (5? C., 5500 rpm, 20 min), and washed once with 0.85% sterile normal saline; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 13% according to a volume ratio of 1:3 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1% according to a volume ratio of 1:0.5 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 30 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 4? C. for later use;
[0100] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (250 rpm, 20 min) evenly with the aqueous solution of the natural polymer material having a concentration of 2% according to a mass ratio of 1:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 111 ?g/g of vitamin A, 4 ?g/g of vitamin D3, 4 mg/g of vitamin E, 8 ?g/g of vitamin K.sub.2, 0.4 mg/g of vitamin B.sub.1, 0.4 mg/g of vitamin B.sub.2, 0.4 mg/g of vitamin B.sub.6, 0.4 ?g/g of vitamin B.sub.12, 4 mg/g of niacinamide, 60 ?g/g of folic acid, 25 mg/g of vitamin C, and 1.5 mg/g of pantothenic acid; the minerals include 113 mg/g of calcium carbonate, 39 mg/g of magnesium gluconate, 0.78 mg/g of manganese sulfate, 3 mg/g of ferrous lactate, 1.5 mg/g of zinc gluconate, 14 ?g/g of sodium selenite, and 0.16 mg/g of copper sulfate; and the prebiotics content was 0.65 g/100 mL;
[0101] Step (4), the solidified probiotics was mixed (350 rpm, 30 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:2 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed evenly with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:6, washed for 3 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0102] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (45 rpm, 15 min) evenly with the wet hydrogel particles according to a mass ratio of 1:15 to obtain a mixture; the mixture was then pre-cooled at ?80? C. for 2.5 h, and then freeze-dried at ?55? C. for 36 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0103] Step (6), an empty capsule shell was fixed and punched with four holes, each having an aperture of 1 mm, in equidistant symmetrical positions at both ends and the waist of the entire shell with laser (as shown in
Example 5
[0104] Step (1), sodium carboxymethyl cellulose (a viscosity of 15000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 310:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:16) to obtain a mixture; the mixture was first stirred at 70 rpm for 100 min, and then stirred at 20 rpm for 14 h to obtain a gel; the gel was dried in a drying oven at 45? C. for 24 h, turned over and continued to dry for 32 h, then cross-linked at high temperature of 130? ? C. for 4.4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed twice with distilled water, 2 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:100 in each washing), and filtered to obtain wet hydrogel particles;
[0105] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 3%, and repeatedly activated for 5 generations under the same culture conditions (38? C., 27 h); bacterial sludge was collected by low-temperature centrifugation (5? C., 5500 rpm, 25 min), and washed twice with 0.9% sterile normal saline; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 20% according to a volume ratio of 1:3 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1.5% according to a volume ratio of 1:1 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 40 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 5? C. for later use;
[0106] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (150 rpm, 30 min) evenly with the aqueous solution of the natural polymer material having a concentration of 4% according to a mass ratio of 1:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 111 ?g/g of vitamin A, 4 ?g/g of vitamin D3, 4 mg/g of vitamin E, 8 ?g/g of vitamin K.sub.2, 0.4 mg/g of vitamin B.sub.1, 0.4 mg/g of vitamin B.sub.2, 0.4 mg/g of vitamin B.sub.6, 0.4 ?g/g of vitamin B.sub.12, 4 mg/g of niacinamide, 60 ?g/g of folic acid, 25 mg/g of vitamin C, and 1.5 mg/g of pantothenic acid; the minerals include 113 mg/g of calcium carbonate, 39 mg/g of magnesium gluconate, 0.78 mg/g of manganese sulfate, 3 mg/g of ferrous lactate, 1.5 mg/g of zinc gluconate, 14 ?g/g of sodium selenite, and 0.16 mg/g of copper sulfate; and the prebiotics content was 0.65 g/100 mL;
[0107] Step (4), the solidified probiotics was mixed (200 rpm, 20 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:3 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:6, washed for 3 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0108] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (60 rpm, 20 min) evenly with the wet hydrogel particles according to a mass ratio of 1:17 to obtain a mixture; the mixture was then pre-cooled at ?80? ? C. for 4 h, and then freeze-dried at ?55? C. for 48 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0109] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 1 mm, in equidistant positions of skew symmetry from the vertex and midline of the entire capsule with laser (as shown in
Example 6
[0110] Step (1), sodium carboxymethyl cellulose (a viscosity of 7000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 330:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:22) to obtain a mixture; the mixture was first stirred at 50 rpm for 80 min, and then stirred at 30 rpm for 16 h to obtain a gel; the gel was dried in a drying oven at 60? C. for 28 h, turned over and continued to dry for 36 h, then cross-linked at high temperature of 110? ? C. for 3.6 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 4 times, 3 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:150 in each washing), and filtered to obtain wet hydrogel particles;
[0111] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 4.5%, and repeatedly activated for 5 generations under the same culture conditions (35? C., 21 h); bacterial sludge was collected by low-temperature centrifugation (4? C., 4500 rpm, 15 min), and washed with 0.95% sterile normal saline for 3 times; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 6% according to a volume ratio of 1:5 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1% according to a volume ratio of 1:1 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 20 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 3?C for later use;
[0112] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (350 rpm, 30 min) evenly with the aqueous solution of the natural polymer material having a concentration of 4% according to a mass ratio of 2:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 128 ?g/g of vitamin A, 6 ?g/g of vitamin D3, 6 mg/g of vitamin E, 10 ?g/g of vitamin K.sub.2, 0.6 mg/g of vitamin B.sub.1, 0.6 mg/g of vitamin B.sub.2, 0.6 mg/g of vitamin B.sub.6, 0.7 ?g/g of vitamin B.sub.12, 7 mg/g of niacinamide, 80 ?g/g of folic acid, 40 mg/g of vitamin C, and 2.5 mg/g of pantothenic acid; the minerals include 133 mg/g of calcium carbonate, 51 mg/g of magnesium gluconate, 0.98 mg/g of manganese sulfate, 5 mg/g of ferrous lactate, 2.5 mg/g of zinc gluconate, 17 ?g/g of sodium selenite, and 0.3 mg/g of copper sulfate; and the prebiotics content was 1.0 g/100 mL;
[0113] Step (4), the solidified probiotics was mixed (300 rpm, 30 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:4 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed evenly with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:4, washed for 4 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0114] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (30 rpm, 10 min) evenly with the wet hydrogel particles according to a mass ratio of 1:13 to obtain a mixture; the mixture was then pre-cooled at ?80? ? C. for 1 h, and then freeze-dried at ?55? C. for 18 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0115] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 1 mm, in symmetrical positions in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Example 7
[0116] Step (1), sodium carboxymethyl cellulose (a viscosity of 11000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 310:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:22) to obtain a mixture; the mixture was first stirred at 60 rpm for 90 min, and then stirred at 20 rpm for 14 h to obtain a gel; the gel was dried in a drying oven at 60? C. for 28 h, turned over and continued to dry for 36 h, then cross-linked at high temperature of 120? C. for 4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed twice with distilled water, 2 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:100 in each washing), and filtered to obtain wet hydrogel particles;
[0117] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 4.5%, and repeatedly activated for 5 generations under the same culture conditions (36.5? C., 24 h); bacterial sludge was collected by low-temperature centrifugation (3? C., 3500 rpm, 10 min), and washed twice with 0.95% sterile normal saline; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 6% according to a volume ratio of 1:5 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1.5% according to a volume ratio of 1:1.5 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 30 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 4? C. for later use;
[0118] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (350 rpm, 30 min) evenly with the aqueous solution of the natural polymer material having a concentration of 4% according to a mass ratio of 1:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 128 ?g/g of vitamin A, 6 ?g/g of vitamin D3, 6 mg/g of vitamin E, 10 ?g/g of vitamin K.sub.2, 0.6 mg/g of vitamin B.sub.1, 0.6 mg/g of vitamin B.sub.2, 0.6 mg/g of vitamin B.sub.6, 0.7 ?g/g of vitamin B.sub.12, 7 mg/g of niacinamide, 80 ?g/g of folic acid, 40 mg/g of vitamin C, and 2.5 mg/g of pantothenic acid; the minerals include 133 mg/g of calcium carbonate, 51 mg/g of magnesium gluconate, 0.98 mg/g of manganese sulfate, 5 mg/g of ferrous lactate, 2.5 mg/g of zinc gluconate, 17 ?g/g of sodium selenite, and 0.3 mg/g of copper sulfate; and the prebiotics content was 1.0 g/100 mL;
[0119] Step (4), the solidified probiotics was mixed (300 rpm, 30 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:4 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed evenly with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:5, washed twice and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0120] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (45 rpm, 15 min) evenly with the wet hydrogel particles according to a mass ratio of 1:13 to obtain a mixture; the mixture was then pre-cooled at ?80? C. for 2.5 h, and then freeze-dried at ?55? C. for 36 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0121] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 0.5 mm, in equidistant symmetrical positions in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Example 8
[0122] Step (1), sodium carboxymethyl cellulose (a viscosity of 15000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 330:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:16) to obtain a mixture; the mixture was first stirred at 50 rpm for 80 min, and then stirred at 30 rpm for 16 h to obtain a gel; the gel was dried in a drying oven at 45? C. for 28 h, turned over and continued to dry for 36 h, then cross-linked at high temperature of 120? C. for 4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 4 times, 3 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:200 in each washing), and filtered to obtain wet hydrogel particles;
[0123] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 3%, and repeatedly activated for 5 generations under the same culture conditions (38? C., 21 h); bacterial sludge was collected by low-temperature centrifugation (4? C., 3500 rpm, 20 min), and washed with 0.9% sterile normal saline for 3 times; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1% according to a volume ratio of 1:1.5 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 30 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a lower temperature of 4? C. for later use;
[0124] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (150 rpm, 30 min) evenly with the aqueous solution of the natural polymer material having a concentration of 2% according to a mass ratio of 2:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 95 ?g/g of vitamin A, 1 ?g/g of vitamin D3, 1 mg/g of vitamin E, 6 ?g/g of vitamin K.sub.2, 0.1 mg/g of vitamin B.sub.1, 0.1 mg/g of vitamin B.sub.2, 0.1 mg/g of vitamin B.sub.6, 0.1 ?g/g of vitamin B.sub.12, 1 mg/g of niacinamide, 40 ?g/g of folic acid, 10 mg/g of vitamin C, and 0.5 mg/g of pantothenic acid; 93 mg/g of calcium carbonate, 27 mg/g of magnesium gluconate, 0.58 mg/g of manganese sulfate, 1 mg/g of ferrous lactate, 0.1 mg/g of zinc gluconate, 10 ?g/g of sodium selenite, and 0.01 mg/g of copper sulfate; and the prebiotics content was 0.3 g/100 mL;
[0125] Step (4), the solidified probiotics was mixed (100 rpm, 30 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:3 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:6, washed for 3 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0126] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (45 rpm, 10 min) evenly with the wet hydrogel particles according to a mass ratio of 1:13 to obtain a mixture; the mixture was then pre-cooled at ?80? C. for 1 h, and then freeze-dried at ?55? C. for 48 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0127] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 0.75 mm, in symmetrical positions in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Example 9
[0128] Step (1), sodium carboxymethyl cellulose (a viscosity of 7000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 350:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:22) to obtain a mixture; the mixture was first stirred at 60 rpm for 90 min, and then stirred at 30 rpm for 16 h to obtain a gel; the gel was dried in a drying oven at 45? C. for 28 h, turned over and continued to dry for 36 h, then cross-linked at high temperature of 110? ? C. for 4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 4 times, 2 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:200 in each washing), and filtered to obtain wet hydrogel particles;
[0129] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 1.5%, and repeatedly activated for 5 generations under the same culture conditions (37? C., 21 h); bacterial sludge was collected by low-temperature centrifugation (5? C., 5500 rpm, 15 min), and washed with 0.85% sterile normal saline for 3 times; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 13% according to a volume ratio of 1:5 to obtain a bacterial suspension; the bacterial suspension was then mixed evenly with an aqueous solution of a natural polymer material having a concentration of 1.5% according to a volume ratio of 1:0.5 to obtain a mixed solution; the obtained mixed solution was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 30 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 3?C for later use;
[0130] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (350 rpm, 10 min) evenly with the aqueous solution of the natural polymer material having a concentration of 4% according to a mass ratio of 1.5:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 95 ?g/g of vitamin A, 1 ?g/g of vitamin D3, 1 mg/g of vitamin E, 6 ?g/g of vitamin K.sub.2, 0.1 mg/g of vitamin B.sub.1, 0.1 mg/g of vitamin B.sub.2, 0.1 mg/g of vitamin B.sub.6, 0.1 ?g/g of vitamin B.sub.12, 1 mg/g of niacinamide, 40 ?g/g of folic acid, 10 mg/g of vitamin C, and 0.5 mg/g of pantothenic acid; the minerals include 93 mg/g of calcium carbonate, 27 mg/g of magnesium gluconate, 0.58 mg/g of manganese sulfate, 1 mg/g of ferrous lactate, 0.1 mg/g of zinc gluconate, 10 ?g/g of sodium selenite, and 0.01 mg/g of copper sulfate; and the prebiotics content was 0.3 g/100 mL;
[0131] Step (4), the solidified probiotics was mixed (300 rpm, 10 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:4 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed with an aqueous solution of an enteric coating material having a concentration of 8% according to a mass ratio of 1:6, washed for 3 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0132] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (60 rpm, 10 min) evenly with the wet hydrogel particles according to a mass ratio of 1:17 to obtain a mixture; the mixture was then pre-cooled at ?80? ? C. for 2.5 h, and then freeze-dried at ?55? C. for 48 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0133] Step (6), an empty capsule shell was fixed and punched with two holes, each having an aperture of 1 mm, in symmetrical positions in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Comparative Example 1
[0134] Step (1), sodium carboxymethyl cellulose (a viscosity of 11000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 330:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:16) to obtain a mixture; the mixture was first stirred at 60 rpm for 90 min, and then stirred at 30 rpm for 16 h to obtain a gel; the gel was dried in a drying oven at 45? C. for 24 h, turned over and continued to dry for 32 h, then cross-linked at high temperature of 120? C. for 4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 4 times, 3 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:150 in each washing), and filtered to obtain wet hydrogel particles;
[0135] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 3%, and repeatedly activated for 5 generations under the same culture conditions (37? C., 24 h); bacterial sludge was collected by low-temperature centrifugation (4? C., 4500 rpm, 15 min), and washed twice with 0.9% sterile normal saline; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 13% according to a volume ratio of 1:4 to obtain a bacterial suspension; the bacterial suspension was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 30 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 4? ? C. for later use;
[0136] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (150 rpm, 20 min) evenly with the aqueous solution of the natural polymer material having a concentration of 3% according to a mass of 1.5:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 111 ?g/g of vitamin A, 4 ?g/g of vitamin D3, 4 mg/g of vitamin E, 8 ?g/g of vitamin K.sub.2, 0.4 mg/g of vitamin B.sub.1, 0.4 mg/g of vitamin B.sub.2, 0.4 mg/g of vitamin B.sub.6, 0.4 ?g/g of vitamin B.sub.12, 4 mg/g of niacinamide, 60 ?g/g of folic acid, 25 mg/g of vitamin C, and 1.5 mg/g of pantothenic acid; the minerals include 113 mg/g of calcium carbonate, 39 mg/g of magnesium gluconate, 0.78 mg/g of manganese sulfate, 3 mg/g of ferrous lactate, 1.5 mg/g of zinc gluconate, 14 ?g/g of sodium selenite, and 0.16 mg/g of copper sulfate; and the prebiotics content was 0.65 g/100 mL;
[0137] Step (4), the solidified probiotics was mixed (200 rpm, 20 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:3 to obtain a mixed nutrient solution;
[0138] Step (5), the mixed nutrient solution was mixed (45 rpm, 15 min) evenly with the wet hydrogel particles according to a mass ratio of 1:15 to obtain a mixture; the mixture was then pre-cooled at ?80? ? C. for 2.5 h, and then freeze-dried at ?55? C. for 36 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0139] Step (6), an empty capsule shell was fixed and punched with four holes, each having an aperture of 1 mm, in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Comparative Example 2
[0140] Step (1), sodium carboxymethyl cellulose (a viscosity of 11000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 330:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:16) to obtain a mixture; the mixture was stirred at 60 rpm for 90 min, and then stirred at 30 rpm for 16 h to obtain a gel; the gel was dried in a drying oven at 45? C. for 24 h, turned over and continued to dry for 32 h, then cross-linked at high temperature of 120? C. for 4 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 4 times, 3 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:150 in each washing), and filtered to obtain wet hydrogel particles;
[0141] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 3%, and repeatedly activated for 5 generations under the same culture conditions (37? C., 24 h); bacterial sludge was collected by low-temperature centrifugation (4? C., 4500 rpm, 15 min), and washed twice with 0.9% sterile normal saline; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 13% according to a volume ratio of 1:4 to obtain a bacterial suspension; the bacterial suspension was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 30 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 4?C for later use;
[0142] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder (the vitamins include vitamin A, vitamin D3, vitamin E, vitamin K.sub.2, vitamin B.sub.1, vitamin B.sub.2, vitamin B.sub.6, vitamin B.sub.12, niacinamide, folic acid, vitamin C, and pantothenic acid; the minerals include calcium carbonate, magnesium gluconate, manganese sulfate, ferrous lactate, zinc gluconate, sodium selenite, and copper sulfate; and the contents of vitamins, minerals and prebiotics in the finished diet-reducing capsule containing multi-nutrient microspheres were the same as those in Example 1);
[0143] Step (4), the vitamin-mineral-prebiotics composite powder, the bacterial suspension and the wet hydrogel particles were mixed (45 rpm, 15 min) evenly according to a mass ratio of 0.5:0.5:15 to obtain a mixture; the mixture was then pre-cooled at ?80? C. for 2.5 h, and then freeze-dried at ?55? C. for 36 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0144] Step (5), an empty capsule shell was fixed and punched with two holes, each having an aperture of 1 mm, in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
Comparative Example 3
[0145] Step (1), sodium carboxymethyl cellulose (a viscosity of 3000) was added to an aqueous solution containing citric acid (a mass ratio of sodium carboxymethyl cellulose to citric acid was 380:1, and a mass ratio of sodium carboxymethyl cellulose to distilled water was 1:32) to obtain a mixture; the mixture was first stirred at 100 rpm for 150 min, and then stirred at 60 rpm for 30 h to obtain a gel; the gel was dried in a drying oven at 75? C. for 30 h, turned over and continued to dry for 40 h, then cross-linked at high temperature of 130? C. for 6 h, crushed and sieved to obtain solid hydrogel particles; the solid hydrogel particles were washed with distilled water for 8 times, 4 h each time (a mass ratio of the solid hydrogel particles to the distilled water was 1:150 in each washing), and filtered to obtain wet hydrogel particles;
[0146] Step (2), probiotics was inoculated in a sterile MRS liquid medium according to an inoculation amount of 7%, and repeatedly activated for 5 generations under the same culture conditions (39? C., 29 h); bacterial sludge was collected by low-temperature centrifugation (6? ? C., 6600 rpm, 30 min), and washed with 0.95% sterile normal saline for 4 times; the washed bacterial sludge was then mixed evenly with an aqueous solution of a freeze-drying protective agent having a concentration of 25% according to a volume ratio of 1:2 to obtain a bacterial suspension; the bacterial suspension was then solidified with a 0.1 mol/L CaCl.sub.2) solution for 50 min, washed and filtered to obtain solidified probiotics; and the solidified probiotics was stored at a low temperature of 6? ? C. for later use;
[0147] Step (3), powders of vitamins, minerals and prebiotics were mixed evenly, and sieved with a 70-mesh sieve to obtain vitamin-mineral-prebiotics composite powder; and the vitamin-mineral-prebiotics composite powder was then mixed (200 rpm, 30 min) evenly with the aqueous solution of the natural polymer material having a concentration of 6% according to a mass ratio of 3:11 to obtain a vitamin-mineral-prebiotics-natural polymer material mixed solution, wherein in the mixed solution, the contents of the vitamins, the minerals and the prebiotics were: 111 ?g/g of vitamin A, 4 ?g/g of vitamin D3, 4 mg/g of vitamin E, 8 ?g/g of vitamin K.sub.2, 0.4 mg/g of vitamin B.sub.1, 0.4 mg/g of vitamin B.sub.2, 0.4 mg/g of vitamin B.sub.6, 0.4 ?g/g of vitamin B.sub.12, 4 mg/g of niacinamide, 60 ?g/g of folic acid, 25 mg/g of vitamin C, and 1.5 mg/g of pantothenic acid; the minerals include 113 mg/g of calcium carbonate, 39 mg/g of magnesium gluconate, 0.78 mg/g of manganese sulfate, 3 mg/g of ferrous lactate, 1.5 mg/g of zinc gluconate, 14 ?g/g of sodium selenite, and 0.16 mg/g of copper sulfate; and the prebiotics content was 0.65 g/100 mL;
[0148] Step (4), the solidified probiotics was mixed (400 rpm, 40 min) evenly with the vitamin-mineral-prebiotics-natural polymer material mixed solution according to a mass ratio of 1:6 to obtain a mixed nutrient solution, and the mixed nutrient solution was then mixed with an aqueous solution of an enteric coating material having a concentration of 2% according to a mass ratio of 1:8, washed for 5 times and filtered to obtain wet probiotics-prebiotics-vitamin-mineral microspheres;
[0149] Step (5), the wet probiotics-prebiotics-vitamin-mineral microspheres were mixed (60 rpm, 40 min) evenly with the wet hydrogel particles according to a mass ratio of 1:20 to obtain a mixture; the mixture was then pre-cooled at ?80? C. for 5 h, and then freeze-dried at ?55? C. for 60 h under a vacuum degree of 25 Pa to obtain a freeze-dried product; the freeze-dried product was then crushed and sieved to obtain a capsule content; and
[0150] Step (6), an empty capsule shell was fixed and punched with four holes, each having an aperture of 1 mm, in the middle of the waist with laser to obtain a porous capsule shell; and the porous capsule shell was then combined with the capsule content to obtain the diet-reducing capsule containing multi-nutrient microspheres.
The Performances of the Diet-Reducing Capsule Containing Multi-Nutrient Microspheres of the Present Disclosure were Measured.
(I) Research on Acid-Resistance Stability Testing of Probiotics in Microspheres
[0151] The samples were placed in simulated gastric juice for 2 h respectively, and then the microspheres were separated, washed and then stored at 4? C. for later use. 1 g of microspheres were added to 9 mL of phosphate buffer solution, then placed in a 37? C. shaker and shaken at 230 rpm for 30 min, followed by sampling, and the total number of viable bacteria was calculated.
[0152] The total number of viable bacteria in probiotics was determined by plate counting, and the specific operation was as follows: activated strains were mixed evenly in an aseptic operation cabinet to obtain a mixture; under the conditions of aseptic operation, the mixture was sequentially diluted into different gradients with sterile water in 10-fold increments; and the diluted products of three suitable dilution gradients were taken respectively, evenly inoculated in an MRS agar solid medium respectively, and invertedly cultured in a constant-temperature incubator at 37? C. for 24 h. Flat plates with the number of colonies between 30 to 300 were counted, and the total number of colonies was calculated according to a formula: total number of colonies (CFU/g)=average number of colonies of the same dilution gradient?dilution factor?5. The test results were shown in Table 1.
(II) Research on Determination of Embedding Rate
[0153] The samples were placed in simulated gastric juice for 2 h, and then microspheres were separated, washed and then stored at 4? C. for later use.
[0154] 1 g of microspheres were added to 9 mL of phosphate buffer solution, shaken in a shaker at 37? C., 230 rpm for 30 min, followed by sampling; the number of viable bacteria was calculated, and the vitamin content, mineral content and prebiotics content were determined. The embedding rate was calculated according to the following formula:
[0156] The test results were shown in Table 2.
(III) Research on Temporal Stability of Vitamins, Minerals, Probiotics and Prebiotics
[0157] The samples were placed at normal temperature and pressure (indoor), and the contents of vitamins, minerals, probiotics and prebiotics in the samples were determined every 2 weeks, lasting for 10 weeks. The determination method of fat-soluble vitamin content refers to the BJS 201717 Determination of 9 Fat-soluble Vitamins in Health Food. The determination method of water-soluble vitamin content refers to the BJS 201716 Determination of 9 Water-soluble Vitamins in Health Food. The determination method of mineral content refers to the BJS 201718 Determination of 9 Mineral Elements in Health Food. The determination method of probiotics content is the same as that in Test (I). The detection method of fructooligosaccharides (prebiotics) refers to GB/T 23528.2-2021 Quality Requirements for oligosaccharides Part 2: Fructooligosaccharides. The test results were shown in Tables 3, 4, 5 and 6.
(IV) Research on Determination of Gel Swelling Rate (MUR).
[0158] A certain mass of samples was weighed (denoted as M.sub.0) and placed in 100 mL of pre-prepared 37? C. diluted artificial gastric juice respectively (a volume ratio of the artificial gastric juice to sterile distilled water was 1:8, pH 2.10), and gently stirred for 0.5 h at a rotation speed of 45 rpm, followed by timing (no bubble was allowed). After the timing was over, excess diluted artificial gastric juice was removed with a stainless steel filter, absorbent paper was used to wipe off the moisture on the surface, and the samples were weighed and counted (denoted as M.sub.1). The MUR was calculated according to a formula. The test was repeated for three samples in parallel, and an average was taken from the results. Formula: MUR=(M.sub.1?M.sub.0)/M.sub.0. The test results were shown in
(V) Research on Determination of Elastic Modulus
[0159] The elasticity of a sample was evaluated by dynamic mechanical analysis (DMA). After the swelling rate determination was over, the sample was immediately placed between parallel plates (with crosshatches and a diameter of 40 mm) of a rotational rheometer, and the elastic modulus was determined. A gap between the steel plates was set to 4 mm. The value of the elastic modulus at a frequency of 10 rad/s was used to determine the elasticity of sample particles. The test was repeated for three samples in parallel, and an average was taken from the results. The test results were shown in
(VI) Research on Effects of Different Concentrations of Enteric Coating Material on the Survival Number of Probiotics in Microspheres
[0160] The samples used in this test were prepared with reference to the method of Example 1, except that the concentration of the enteric coating material was changed according to
[0161] The samples (different concentrations of enteric coating material) were placed in sterile water for 2 h, and then microspheres were separated, washed and then stored at 4? C. for later use. 1 g of microspheres were added to 9 mL of phosphate buffer solution, then placed in a 37? C. shaker and shaken at 230 rpm for 30 min, followed by sampling, and the total number of viable bacteria was calculated.
[0162] The determination method for the total number of viable bacteria in probiotics was the same as that in Test (I). The test results were shown in
(VII) Research on Effects of the Length of Time the Microspheres Stay in the Intestine on the Survival Number of Probiotics in the Microspheres
[0163] The samples used in this test were the samples prepared in Example 1.
[0164] The samples were placed in sterile water for 2 h, and then the microspheres were separated, washed and stored at 4? C. for later use. 1 g of microspheres were added to 9 containers each containing 9 mL of artificial intestinal fluid. The containers were then placed in a 37? C. shaker and shaken at 230 rpm for different times (0 min, 15 min, 30 min, 45 min, 60 min, 75 min, 90 min, 105 min, and 120 min), followed by sampling. The total number of viable bacteria was calculated.
[0165] The determination method for the total number of viable bacteria in probiotics was the same as that in Test (I). The test results were shown in
(VIII) Research on Effects of Different Laser Punching Methods on Determination of the Disintegration Time of Capsule
[0166] The samples used in this test were prepared with reference to the method of Example 1, except that the empty capsule shell was punched with different laser punching methods, and other conditions were the same as those of Example 1.
[0167] A test design for the effects of different laser punching methods on determination of the disintegration time of a capsule was shown in Table 7.
TABLE-US-00001 TABLE 7 Test Number of Aperture code Punching positions on empty capsule shell punching (hole) (mm) A Symmetrical positions in the middle of the waist 2 1.00 B Symmetrical positions in the middle of the waist 4 1.00 C Central symmetrical positions at both ends 2 1.00 D Equidistant symmetrical positions at both ends and 4 1.00 the waist E Equidistant positions of skew symmetry from the 2 1.00 vertex and midline F Symmetrical positions in the middle of the waist 2 0.75 G Symmetrical positions in the middle of the waist 4 0.75 H Central symmetrical positions at both ends 2 0.75 I Equidistant symmetrical positions at both ends and 4 0.75 the waist J Equidistant positions of skew symmetry from the 2 0.75 vertex and midline K Symmetrical positions in the middle of the waist 2 0.50 L Symmetrical positions in the middle of the waist 4 0.50 M Central symmetrical positions at both ends 2 0.50 N Equidistant symmetrical positions at both ends and 4 0.50 the waist O Equidistant positions of skew symmetry from the 2 0.50 vertex and midline
[0168] Capsule disintegration time determination method: the test was carried out in accordance with the Pharmacopoeia of the People's Republic of China, Edition 2020, Part IV, General Principles 0921, Disintegration Time Limit Inspection Method. Six test sample capsules were taken, a disintegration time limit tester was checked and equipped with a baffle, the temperature was set to 37? ? C., and the time when the capsules were completely dissolved in the artificial gastric juice was recorded. The capsules should be completely disintegrated within 30 min. If there was one capsule that cannot be completely disintegrated, another six capsules should be taken for retesting. If there was another one capsule that cannot be completely disintegrate, it would be recorded as unqualified. The test was repeated for three samples in parallel, and an average was taken from the results. The test results were shown in
(IX) Study of Weight Loss Effect of Capsule Prepared by the Present Disclosure
[0169] Example 1 was selected as a test sample. Five male and female obese rats of the same age and sexual maturity were taken respectively, with a male rat weight of 500 to 520 g and a female rat weight of 400 to 420 g. The obese rats were given one capsule 30 min before meals every day at regular time, the obese rats were given normal diet after 30 min, and the test continued in this way for 63 days; and the weights of the obese rats were weighed and recorded at regular time of 20:00 every 7 days. The test results were shown in
Performance Test Results of Examples 1 to 9 and Comparative Examples 1 to 3 as Samples were Described as Follows
[0170]
TABLE-US-00002 TABLE 1 Research results of acid resistance of probiotics in microspheres Samples Acid resistance of probiotics (CPU/g) Example 1 2.22 ? 10.sup.8 Example 2 1.59 ? 10.sup.8 Example 3 1.85 ? 10.sup.8 Example 4 1.88 ? 10.sup.8 Example 5 1.46 ? 10.sup.8 Example 6 1.63 ? 10.sup.8 Example 7 1.73 ? 10.sup.8 Example 8 1.64 ? 10.sup.8 Example 9 1.53 ? 10.sup.8 Comparative example 1 4.2 ? 10.sup.5 Comparative example 2 5.6 ? 10.sup.4 Comparative example 3 1.3 ? 10.sup.7
TABLE-US-00003 TABLE 2 Embedding rates of probiotics, vitamins, minerals and prebiotics in microspheres Probiotics Vitamins Minerals Prebiotics Sample (%) (%) (%) (%) Example 1 87.8 86.98 81.12 86.15 Example 2 84.65 81.16 75.32 77 Example 3 86.61 84.92 73.33 76.67 Example 4 85.83 80.15 79.78 78.46 Example 5 83.86 82.74 78.04 81.54 Example 6 83.86 82.79 78.46 75 Example 7 85.04 81.69 75.12 77 Example 8 84.65 80.13 80.79 83.33 Example 9 83.07 80.44 77.21 73.33 Comparative 83.86 78.45 70.66 76.92 example 1 Comparative 30.71 60.11 41.75 36.92 example 2 Comparative 85.43 81.4 60.86 69.23 example 3
TABLE-US-00004 TABLE 3 Storage stability of probiotics in microspheres Storage stability of probiotics (?10.sup.8 CPU/g) Sample Week 0 Week 2 Week 4 Week 6 Week 8 Week 10 Example 1 2.23 2.31 2.26 2.21 2.18 2.14 Example 2 2.15 2.09 2.05 2.01 1.97 1.94 Example 3 2.19 2.16 2.11 2.07 2.05 2.01 Example 4 2.18 2.15 2.11 2.07 2.03 1.98 Example 5 2.13 2.08 2.03 1.99 1.95 1.91 Example 6 2.13 2.09 2.05 2.01 1.97 1.92 Example 7 2.16 2.11 2.02 1.96 1.89 1.86 Example 8 2.15 2.09 2.04 2.01 1.94 1.87 Example 9 2.11 2.06 1.99 1.93 1.87 1.79 Comparative 2.24 2.2 2.15 2.11 2.06 2.01 example 1 Comparative 2.17 2.02 1.86 1.68 1.56 1.37 example 2 Comparative 1.84 1.81 1.78 1.71 1.65 1.62 example 3
TABLE-US-00005 TABLE 4 Storage stability of vitamins in microspheres Storage stability of vitamins in microspheres (%) Sample Week 0 Week 2 Week 4 Week 6 Week 8 Week 10 Example 1 86.98 86.73 86.51 86.28 86.01 85.81 Example 2 81.16 80.88 80.55 80.13 79.5 79.02 Example 3 84.92 84.56 84.09 83.68 83.48 83.25 Example 4 80.15 79.83 79.43 79.14 78.88 78.62 Example 5 82.74 82.72 82.51 82.29 81.82 81.82 Example 6 82.79 82.62 82.31 81.98 81.68 81.45 Example 7 81.69 81.57 81.07 80.68 80.24 79.91 Example 8 80.13 79.96 79.74 79.52 79.23 78.96 Example 9 80.44 80.31 80.09 79.76 79.35 79.01 Comparative 78.45 77.32 76.62 75.38 74.52 73.45 example 1 Comparative 60.11 58.87 56.69 54.53 52.46 51.34 example 2 Comparative 81.4 80.97 80.51 80.15 79.85 79.28 example 3
TABLE-US-00006 TABLE 5 Storage stability of minerals in microspheres Storage stability of minerals in microspheres (%) Sample Week 0 Week 2 Week 4 Week 6 Week 8 Week 10 Example 1 81.12 81.07 80.84 80.62 80.48 80.24 Example 2 75.32 75.31 74.76 74.52 74.12 73.71 Example 3 73.33 73.32 72.83 72.37 71.85 71.49 Example 4 79.78 79.66 79.52 79.29 79.14 78.85 Example 5 78.04 77.94 77.55 77.19 76.81 76.45 Example 6 78.46 78.34 78.05 77.69 77.35 77.14 Example 7 75.12 74.97 74.64 74.48 74.25 73.85 Example 8 80.79 80.56 80.14 80.03 79.81 79.62 Example 9 77.21 77.04 76.61 76.24 79.81 75.48 Comparative 70.66 68.54 67.41 66.26 65.25 63.89 example 1 Comparative 41.75 39.96 37.61 34.56 32.24 30.24 example 2 Comparative 60.86 59.62 59.18 58.61 58.23 57.58 example 3
TABLE-US-00007 TABLE 6 Storage stability of prebiotics in microspheres Storage stability of prebiotics in microspheres (%) Sample Week 0 Week 2 Week 4 Week 6 Week 8 Week 10 Example 1 86.15 86.22 85.96 85.61 85.29 84.97 Example 2 77.02 76.77 76.43 76.26 75.92 75.34 Example 3 76.67 76.42 76.05 75.68 75.19 74.77 Example 4 78.46 78.31 78.05 77.72 77.19 76.87 Example 5 81.54 81.23 80.99 80.64 80.34 79.92 Example 6 75.01 74.97 74.72 74.37 74.13 73.96 Example 7 77.04 76.98 76.65 76.29 76.14 75.91 Example 8 83.33 82.97 82.51 82.24 81.84 81.33 Example 9 73.33 72.99 72.67 72.24 71.75 71.59 Comparative 76.92 75.94 75.51 75.04 74.25 73.71 example 1 Comparative 36.92 34.52 33.14 30.66 29.44 28.24 example 2 Comparative 69.23 68.84 68.64 68.25 67.48 67.06 example 3
[0171] As can be seen from Table 1, compared with Comparative examples 1 to 3, the number of viable bacteria in each of Examples 1 to 9 was more than 10.sup.8, indicating that the probiotics in the microspheres had good acid resistance. As can be seen from Table 2, in Examples 1 to 9, the embedding rates of probiotics and vitamins were more than 80% respectively, the embedding rate of minerals was more than 70%, the embedding rate of prebiotics was more than 75%, and the embedding rate of nutrients in the microspheres was good. As can be seen from Tables 3, 4, 5 and 6, the probiotics, vitamins, minerals and prebiotics had good storage stability. As can be seen from
[0172] Although the contents of the present disclosure have been described in detail by the preferred examples above, it should be recognized that the above description should not be considered as a limitation on the present disclosure. After a person skilled in the art has read the above contents, a variety of modifications and substitutions made for the present disclosure will be obvious. Therefore, the protection scope of the present disclosure should be defined by the appended claims.