CN114392282B - Preparation with high metazoan content and preparation method and application thereof - Google Patents
Preparation with high metazoan content and preparation method and application thereof Download PDFInfo
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/40—Cyclodextrins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Abstract
The invention belongs to the technical field of biological preparations, and particularly relates to a preparation with high metazoan content, and a preparation method and application thereof. The preparation with high metazoan content comprises the following components in parts by weight: 60-70 parts of metazoan, 0.5-1 part of polyvinyl alcohol, 2-5 parts of absorption accelerator and 0.1-0.3 part of stabilizer; the absorption enhancer comprises sorbitol and L-fucose; the stabilizing agent comprises polyvinylpyrrolidone and hydroxypropyl cyclodextrin. The preparation provided by the invention has high postnatal vitality content and excellent anti-immunosenescence effect. In addition, the preparation method of the preparation with high metazoan content is simple, easy to implement industrially and beneficial to large-scale popularization and application.
Description
Technical Field
The invention belongs to the technical field of biological preparations, and particularly relates to a preparation with high metazoan content, and a preparation method and application thereof.
Background
Metazoa refers to inactivated probiotic cells and their associated metabolic byproducts, having the ability to provide a benefit to the health of the host, and has been defined broadly as: metazoan are non-living microorganisms and/or components thereof (including metabolites, cellular components, or cell-free mixtures) that are beneficial to the health of the host; including many different components such as short chain fatty acids, microbial cell components, exopolysaccharides, cell lysates, teichoic acids, peptidoglycans, active lipids, organic acids, and the like. Compared with live probiotics, metazoans have clear chemical structures, safe dosage parameters, longer shelf life and good absorption, metabolism, distribution and excretion capacities, and simultaneously overcome the potential risks of probiotics in the use process, such as abdominal distension and flatulence, probiotics-related bacteremia and mycosis, and possible risks of antibiotic resistance gene drift and the like. Therefore, the development of the metazoan has wide market prospect.
At present, the development of the postbiotic preparation mainly focuses on improving the intestinal health, losing weight and reducing fat, and for example, the Chinese patent application with the publication number of CN111494607A discloses a selenium-rich postbiotic fermentation traditional Chinese medicine microbial preparation for improving the growth performance of piglets and improving the intestinal health, which comprises the following components in parts by weight: 5-10 parts of Chinese pulsatilla root, 10-20 parts of astragalus membranaceus, 5-10 parts of radix sophorae flavescentis, 10-20 parts of Chinese yam, 10-20 parts of Chinese angelica, 10-20 parts of hawthorn, 6-8 parts of antibacterial peptide, 8-10 parts of selenium-enriched prebiotics, 6-10 parts of saccharomycetes, 6-10 parts of lactobacillus, 1000-1200 parts of soybean meal and 500-700 parts of corn flour. Chinese patent application publication No. CN113854461A discloses a composite post-biotic solid beverage, which is composed of sulforaphane, sodium butyrate, post-biotic solid powder, binder, sweetener and sour agent; the solid beverage is especially suitable for people with special requirements such as hyperlipemia, fatty liver, and weight reducing people.
The preparations containing metazoan on the market at present have low content of metazoan in the preparations and few metazoan preparations having anti-immunosenescence effect, so that the development of a metazoan preparation having high content of metazoan and excellent anti-immunosenescence effect is urgent.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a preparation with high metazoan content, a preparation method and application thereof. The preparation provided by the invention has high postnatal vitality content and excellent anti-immunosenescence effect. In addition, the preparation method of the preparation with high metazoan content provided by the invention is simple, is easy for industrial implementation, and is beneficial to large-scale popularization and application.
The technical scheme of the invention is as follows:
a preparation with high metazoan content comprises the following components in parts by mass:
60-70 parts of metazoan, 0.5-1 part of polyvinyl alcohol, 2-5 parts of absorption accelerator and 0.1-0.3 part of stabilizer; the absorption enhancer comprises sorbitol and L-fucose; the stabilizer comprises polyvinylpyrrolidone and hydroxypropyl cyclodextrin.
Further, the preparation with high metazoan content comprises the following components in parts by mass:
64 parts of metazoan, 0.7 part of polyvinyl alcohol, 3 parts of absorption accelerator and 0.2 part of stabilizer.
Further, the absorption enhancer is prepared from sorbitol and L-fucose in a mass ratio of 4-6: 1.
Further, the absorption enhancer is prepared from sorbitol and L-fucose according to a mass ratio of 5: 1.
Further, the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 2-4: 10-13.
Further, the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 3: 10.
Further, the preparation method of the metazoan comprises the following steps:
(1) adding water into the fermentation material, wherein the adding amount of the water is 10-20 times of the mass of the fermentation material, stirring at the stirring speed of 600-;
(2) adding fermentation bacteria into the mixed material obtained in the step (1), wherein the adding amount of the fermentation bacteria is 1-3% of the mass of the mixed material, fermenting, adjusting the fermentation temperature to 27-33 ℃, the fermentation time to 4-7h, adjusting the fermentation temperature to 34-36 ℃ after the fermentation is finished, and fermenting again, wherein the fermentation time is 5-10h to obtain a fermented material;
(3) filtering the fermentation material obtained in the step (2), sterilizing and centrifuging the filtrate, wherein the centrifugation speed is 2000-3000r/min, the centrifugation time is 10-20min, and collecting the supernatant.
Further, the fermentation material in the step (1) of the preparation method of the metazoan comprises the following raw materials in parts by weight:
20-30 parts of glucose, 6-12 parts of peptone, 0.2-0.5 part of tremella polysaccharide, 8-12 parts of jerusalem artichoke powder, 1-3 parts of dipotassium hydrogen phosphate and 0.4-0.8 part of magnesium sulfate.
Further, the fermentation material comprises the following raw materials in parts by weight:
26 parts of glucose, 9 parts of peptone, 0.3 part of tremella polysaccharide, 10 parts of jerusalem artichoke powder, 2 parts of dipotassium hydrogen phosphate and 0.6 part of magnesium sulfate.
Further, the preparation method of the anagen comprises the step (2) that the zymocyte consists of lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subsp.
Further, the fermentation bacteria are prepared from lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subsp. infantis according to a mass ratio of 10-15: 6-8: 2-3.
Further, the fermentation bacteria are prepared from lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subsp. infantis in a mass ratio of 12: 7: 3, the components are mixed.
In the invention, the CAS number of the tremella polysaccharide: 9075-53-0.
The preparation method of the preparation with high metazoan content comprises the following steps:
taking the latter biogen, adding polyvinyl alcohol at the stirring speed of 300-500r/min, stirring for 25-40min, adding the absorption promoter, continuing stirring for 30-50min, adding the stabilizer, and continuing stirring for 1-2 h.
The invention also aims to provide the application of the preparation with high metazoan content in preparing the anti-immunosenescence medicament.
The preparation with high content of the metaplasia provided by the invention can generate a synergistic effect among the components by reasonably setting the components and the content of the metaplasia, the polyvinyl alcohol, the absorption enhancer and the stabilizer, is favorable for the better effect of the active ingredients of the metaplasia in the obtained preparation, has excellent stability and is favorable for the stable storage of the preparation. The absorption enhancer composed of the sorbitol and the L-fucose according to a specific ratio can not only promote the absorption of the metazoan effective components, but also generate a synergistic effect with the metazoan effective components, and exert more excellent anti-immunosenescence effect.
The postnatal preparation is prepared by the steps of firstly adding water into a fermentation material, stirring to obtain a mixed material, adding fermentation bacteria, fermenting and the like. The added fermentation bacteria composed of lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subspecies infantis according to a certain mass ratio act together, so that the fermentation speed can be increased, the fermentation time can be shortened, and the post-growth effective components such as lipoteichoic acid and the like can be obtained. The tremella polysaccharide added in the fermentation material is beneficial to the culture of thalli, and can produce more post-growth effective components such as lipoteichoic acid and the like. The invention firstly ferments for a certain time at a certain temperature, and then regulates the fermentation temperature to ferment for a certain time again, which is beneficial to the exertion of the effect of the strain and the mass acquisition of the metazoan. The operations in the invention act together to finally achieve the purpose of improving the content of the metazoan.
Compared with the prior art, the invention has the following advantages:
(1) the preparation prepared from the prebiotics, the polyvinyl alcohol, the absorption enhancer and the stabilizer has high prebiotic content and excellent anti-immunosenescence effect.
(2) The preparation method of the preparation with high metazoan content is simple, is easy for industrial implementation, and is beneficial to large-scale popularization and application.
(3) The preparation with high metazoan content provided by the invention has excellent stability, is beneficial to stable storage of the preparation, has longer shelf life, and is easy to store and transport.
Detailed Description
The present invention is further described in the following description of the specific embodiments, which is not intended to limit the invention, but various modifications and improvements can be made by those skilled in the art according to the basic idea of the invention, within the scope of the invention, as long as they do not depart from the basic idea of the invention.
The starting materials used in the present invention are all commercially available unless otherwise specified. If the lactobacillus casei can be purchased from the Guangdong province microorganism strain preservation center, the strain preservation number is as follows: GDMCC 1.204, lactobacillus plantarum strain accession number: GDMCC 1.191, bifidobacterium longum subspecies infantis, strain deposit No.: GDMCC 1.207.
Example 1A high metazoan formulation
The preparation with high metazoan content comprises the following components in parts by mass:
60 parts of postbiotic, 0.5 part of polyvinyl alcohol, 2 parts of absorption promoter and 0.1 part of stabilizer; the absorption enhancer is prepared from sorbitol and L-fucose in a mass ratio of 4: 1, preparing a composition; the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 2: 13.
The preparation method of the anagen comprises the following steps:
(1) adding water into the fermented material, wherein the adding amount of the water is 10 times of the mass of the fermented material, and stirring at the stirring speed of 600r/min for 20min to obtain a mixed material;
(2) adding zymocyte into the mixed material obtained in the step (1), wherein the zymocyte is prepared from lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subspecies neonatorum according to the mass ratio of 10: 8: 3, fermenting, wherein the adding amount of the zymophyte is 1 percent of the mass of the mixed material, the fermentation temperature is adjusted to be 27 ℃, the fermentation time is 4 hours, the fermentation temperature is adjusted to be 34 ℃ after the fermentation is finished, and secondary fermentation is carried out for 5 hours to obtain a fermented material;
(3) and (3) filtering the fermented material obtained in the step (2), sterilizing the filtrate, centrifuging at the speed of 2000r/min for 10min, and collecting the supernatant to obtain the fermented material.
The fermentation material comprises the following raw materials in parts by weight:
20 parts of glucose, 6 parts of peptone, 0.2 part of tremella polysaccharide, 8 parts of jerusalem artichoke powder, 1 part of dipotassium hydrogen phosphate and 0.4 part of magnesium sulfate.
The preparation method of the preparation with high metazoan content comprises the following steps:
taking the postnatal, adding polyvinyl alcohol at the stirring speed of 300r/min, stirring for 25min, adding the absorption enhancer, continuing stirring for 30min, adding the stabilizer, and continuing stirring for 1h to obtain the final product.
Example 2A high metazoan formulation
The preparation with high metazoan content comprises the following components in parts by mass:
70 parts of postbiotic, 1 part of polyvinyl alcohol, 5 parts of absorption accelerator and 0.3 part of stabilizer; the absorption enhancer is prepared from sorbitol and L-fucose according to a mass ratio of 6: 1, preparing a composition; the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 4: 10.
The preparation method of the anagen comprises the following steps:
(1) adding water into the fermented material, wherein the adding amount of the water is 20 times of the mass of the fermented material, and stirring at the stirring speed of 900r/min for 40min to obtain a mixed material;
(2) adding zymophyte into the mixed material obtained in the step (1), wherein the zymophyte is prepared from lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subspecies neonatorum according to a mass ratio of 15: 6: 2, fermenting, wherein the adding amount of the zymophyte is 3% of the mass of the mixed material, the fermentation temperature is adjusted to 33 ℃, the fermentation time is 7 hours, the fermentation temperature is adjusted to 36 ℃ after the fermentation is finished, and secondary fermentation is carried out for 10 hours to obtain a fermented material;
(3) and (3) filtering the fermented material obtained in the step (2), sterilizing and centrifuging the filtrate at the centrifugation speed of 3000r/min for 20min, and collecting the supernatant to obtain the fermented material.
The fermentation material comprises the following raw materials in parts by weight:
30 parts of glucose, 12 parts of peptone, 0.5 part of tremella polysaccharide, 12 parts of jerusalem artichoke powder, 3 parts of dipotassium hydrogen phosphate and 0.8 part of magnesium sulfate.
The preparation method of the preparation with high metazoan content comprises the following steps:
taking the later-born material, adding polyvinyl alcohol at a stirring speed of 500r/min, stirring for 40min, adding the absorption enhancer, continuing to stir for 50min, adding the stabilizer, and continuing to stir for 2h to obtain the final product.
Example 3A high metazoan formulation
The preparation with high metazoan content comprises the following components in parts by mass:
64 parts of metazoan, 0.7 part of polyvinyl alcohol, 3 parts of absorption accelerator and 0.2 part of stabilizer; the absorption enhancer is prepared from sorbitol and L-fucose in a mass ratio of 5: 1, preparing a composition; the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 3: 10.
The preparation method of the anagen comprises the following steps:
(1) adding water into the fermentation material, wherein the adding amount of the water is 14 times of the mass of the fermentation material, and stirring at the stirring speed of 800r/min for 25min to obtain a mixed material;
(2) adding zymocyte into the mixed material obtained in the step (1), wherein the zymocyte is prepared from lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subspecies neonatorum according to the mass ratio of 12: 7: 3, fermenting, wherein the adding amount of the zymophyte is 1.5 percent of the mass of the mixed material, the fermentation temperature is adjusted to be 30 ℃, the fermentation time is 5 hours, the fermentation temperature is adjusted to be 35 ℃ after the fermentation is finished, and secondary fermentation is carried out for 8 hours to obtain a fermented material;
(3) and (3) filtering the fermented material obtained in the step (2), sterilizing the filtrate, centrifuging at the speed of 2800r/min for 18min, and collecting the supernatant to obtain the fermented material.
The fermentation material comprises the following raw materials in parts by weight:
26 parts of glucose, 9 parts of peptone, 0.3 part of tremella polysaccharide, 10 parts of jerusalem artichoke powder, 2 parts of dipotassium hydrogen phosphate and 0.6 part of magnesium sulfate.
The preparation method of the preparation with high metazoan content comprises the following steps:
taking the latter prebiotics, adding polyvinyl alcohol at a stirring speed of 400r/min, stirring for 35min, adding the absorption enhancer, continuing to stir for 40min, adding the stabilizer, and continuing to stir for 1.5h to obtain the product.
Comparative example 1A preparation containing metazoan
The preparation containing the metazoan comprises the following components in parts by weight:
64 parts of postbiotic, 0.7 part of polyvinyl alcohol, 3 parts of absorption promoter and 0.2 part of stabilizer; the absorption enhancer is prepared from sorbitol and trehalose according to a mass ratio of 5: 1, preparing a composition; the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 3: 10.
The method for preparing the anagen and the preparation containing the anagen is similar to the example 3.
The difference from example 3 is that L-fucose is replaced by trehalose.
Comparative example 2A preparation containing prebiotics
The preparation containing the metazoan comprises the following components in parts by weight:
64 parts of postbiotic, 0.7 part of polyvinyl alcohol, 3 parts of absorption promoter and 0.2 part of stabilizer; the absorption enhancer is prepared from sorbitol and L-fucose in a mass ratio of 5: 1, preparing a composition; the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 3: 10.
The preparation method of the metancholia comprises the following steps:
(1) adding water into the fermented material, wherein the adding amount of the water is 14 times of the mass of the fermented material, and stirring at the stirring speed of 800r/min for 25min to obtain a mixed material;
(2) adding zymocyte into the mixed material obtained in the step (1), wherein the zymocyte is prepared by mixing lactobacillus casei and lactobacillus plantarum according to a mass ratio of 12: 7, fermenting, wherein the adding amount of the zymophyte is 1.5 percent of the mass of the mixed material, the fermentation temperature is adjusted to be 30 ℃, the fermentation time is 5 hours, the fermentation temperature is adjusted to be 35 ℃ after the fermentation is finished, and secondary fermentation is carried out for 8 hours to obtain a fermented material;
(3) and (3) filtering the fermentation material obtained in the step (2), sterilizing and centrifuging the filtrate at the speed of 2800r/min for 18min, and collecting the supernatant to obtain the fermented soybean protein.
The fermentation material comprises the following raw materials in parts by weight:
26 parts of glucose, 9 parts of peptone, 0.3 part of tremella polysaccharide, 10 parts of jerusalem artichoke powder, 2 parts of dipotassium hydrogen phosphate and 0.6 part of magnesium sulfate.
The preparation method of the preparation containing the metazoan is similar to that of the example 3.
The difference from example 3 is that the fermentation tubes are not added with bifidobacterium longum subsp.
Comparative example 3A preparation containing prebiotics
The preparation containing the metazoan comprises the following components in parts by weight:
64 parts of metazoan, 0.7 part of polyvinyl alcohol, 3 parts of absorption accelerator and 0.2 part of stabilizer; the absorption enhancer is prepared from sorbitol and L-fucose in a mass ratio of 5: 1, preparing a composition; the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 3: 10.
The preparation method of the metancholia comprises the following steps:
(1) adding water into the fermentation material, wherein the adding amount of the water is 14 times of the mass of the fermentation material, and stirring at the stirring speed of 800r/min for 25min to obtain a mixed material;
(2) adding zymophyte into the mixed material obtained in the step (1), wherein the zymophyte is prepared from lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subspecies infantis according to the mass ratio of 12: 7: 3, adding the zymophyte in an amount of 1.5 percent of the mass of the mixed material, fermenting, adjusting the fermentation temperature to 30 ℃, and fermenting for 13 hours to obtain a fermented material;
(3) and (3) filtering the fermentation material obtained in the step (2), sterilizing and centrifuging the filtrate at the speed of 2800r/min for 18min, and collecting the supernatant to obtain the fermented soybean protein.
The fermentation material comprises the following raw materials in parts by weight:
26 parts of glucose, 9 parts of peptone, 0.3 part of tremella polysaccharide, 10 parts of jerusalem artichoke powder, 2 parts of dipotassium hydrogen phosphate and 0.6 part of magnesium sulfate.
The preparation method of the preparation containing the metazoan is similar to that of the example 3.
The difference from example 3 is that the specific fermentation operation in step (2) of the method for the preparation of metazoan is different.
Comparative example 4A preparation containing metazoan
The preparation containing the metazoan comprises the following components in parts by weight:
64 parts of postbiotic, 0.7 part of polyvinyl alcohol, 3 parts of absorption promoter and 0.2 part of stabilizer; the absorption enhancer is prepared from sorbitol and L-fucose in a mass ratio of 5: 1, preparing a composition; the stabilizer is prepared from polyvinylpyrrolidone and hydroxypropyl cyclodextrin in a mass ratio of 3: 10.
The preparation method of the metancholia comprises the following steps:
(1) adding water into the fermented material, wherein the adding amount of the water is 14 times of the mass of the fermented material, and stirring at the stirring speed of 800r/min for 25min to obtain a mixed material;
(2) adding zymocyte into the mixed material obtained in the step (1), wherein the zymocyte is prepared from lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subspecies neonatorum according to the mass ratio of 12: 7: 3, adding zymophyte in an amount of 1.5 percent of the mass of the mixed material, fermenting, adjusting the fermentation temperature to be 30 ℃, the fermentation time to be 5 hours, adjusting the fermentation temperature to be 35 ℃ after the fermentation is finished, and fermenting again for 8 hours to obtain a fermented material;
(3) and (3) filtering the fermentation material obtained in the step (2), sterilizing and centrifuging the filtrate at the speed of 2800r/min for 18min, and collecting the supernatant to obtain the fermented soybean protein.
The fermentation material comprises the following raw materials in parts by weight:
26 parts of glucose, 9 parts of peptone, 10 parts of jerusalem artichoke powder, 2 parts of dipotassium hydrogen phosphate and 0.6 part of magnesium sulfate.
The preparation method of the preparation containing the metazoan is similar to that of the example 3.
The difference from example 3 is that no tremella polysaccharide is added to the fermentation material.
Test example I measurement of Lipoteichoic acid content
1. Test materials: example 3, comparative examples 2 to 4.
2. The test method comprises the following steps: the content of lipoteichoic acid in the metazoan obtained in example 3 and comparative examples 2 to 4 was measured using LTA lipoteichoic acid elisa kit from Shanghai Biotechnology Ltd.
3. And (3) test results: the specific measurement results are shown in table 1.
Table 1: measurement of lipoteichoic acid content
| Measurement items | Example 3 | Comparative example 2 | Comparative example 3 | Comparative example 4 |
| Lipoteichoic acid (pg/mL) | 352.9 | 263.7 | 305.4 | 291.2 |
As can be seen from Table 1, the amount of lipoteichoic acid in the after-growth starter obtained by the method of example 3 according to the present invention is significantly higher than that of the after-growth starter obtained by comparative examples 2-4. Therefore, the content of lipoteichoic acid in the anagen obtained by the method is higher, and a foundation is laid for preparing a preparation with high anagen content.
Test example two test for anti-immunosenescence Effect
1. Test materials: the high metazoan content formulations prepared in example 3 and comparative examples 1-4.
2. Test subjects: kunming-line mice, 2 months old, 20g in weight, 42 mice, half male and female, provided by Shanghai laboratory animal center of Chinese academy of sciences, and production license numbers: SCXK (Shanghai) 2003-0003.
3. The test method comprises the following steps:
the mice were randomly divided into example 3, model, control, comparative 1, comparative 2, comparative 3, and comparative 4 groups, each group having 6 mice each with half of male and female. D-galactose was diluted with 0.9% physiological saline to prepare a 5% (50mg/mL) D-galactose physiological saline solution. The model group is injected with 0.25mL/10g of 5% D-galactose physiological saline solution subcutaneously on the back of the neck once a day, and is injected with 0.3 mL/mouse of sterile physiological saline solution in the abdominal cavity; the group of example 3 and the group of comparative examples 1 to 4 were intraperitoneally injected with the preparations with high metazoan content prepared in example 3 and the preparations with high metazoan content prepared in comparative examples 1 to 4, respectively, 100 μ g/mouse, and the control group was injected with equal volume of sterile physiological saline once a day under the dorsal nuchal and intraperitoneally simultaneously. Each group of mice was weighed after continuous treatment for 40 days, killed by neck-amputation, thymus was taken out, and after residual blood was blotted with filter paper, weighing was performed, and thymus index was calculated from the corresponding body weight. Thymus index is thymus weight (mg)/body weight (g). The mouse is removed from the eyeball to take blood, serum is separated conventionally, and the IL-2 content in peripheral blood is determined.
4. And (3) test results: the effect of the formulation on thymus index, peripheral blood IL-2 content is shown in Table 2.
Table 2: effect of the preparation on thymic index
As can be seen from Table 2, the thymus index and IL-2 content of the mice in the model group were significantly reduced compared to the control group, indicating that the molding was successful. Compared with the model group, the thymus index and the IL-2 content of the mice in the groups of example 3 and comparative examples 1-4 are improved to a certain extent after 40 days of continuous treatment, wherein the thymus index and the IL-2 content of the mice in the group of example 3 are obviously improved and are closer to those of the control group. Therefore, the preparation with high metazoan content has obvious anti-immunosenescence effect.
Claims (6)
1. The preparation containing the metancholia is characterized by comprising the following components in parts by weight:
60-70 parts of metazoan, 0.5-1 part of polyvinyl alcohol, 2-5 parts of absorption accelerator and 0.1-0.3 part of stabilizer; the absorption enhancer comprises sorbitol and L-fucose; the stabilizing agent comprises polyvinylpyrrolidone and hydroxypropyl cyclodextrin;
the preparation method of the metancholia comprises the following steps:
(1) adding water into the fermentation material, wherein the adding amount of the water is 10-20 times of the mass of the fermentation material, stirring at the stirring speed of 600-900r/min for 20-40min to obtain a mixed material;
(2) adding zymophyte into the mixed material obtained in the step (1), wherein the adding amount of the zymophyte is 1-3% of the mass of the mixed material, fermenting, adjusting the fermentation temperature to be 27-33 ℃, the fermentation time to be 4-7h, adjusting the fermentation temperature to be 34-36 ℃ after the fermentation is finished, and fermenting again, wherein the fermentation time is 5-10h to obtain a fermented material;
(3) filtering the fermentation material obtained in the step (2), sterilizing and centrifuging the filtrate, wherein the centrifugation speed is 2000-;
the preparation method of the metazoan comprises the following steps of (1) preparing the fermentation material, wherein the fermentation material comprises the following raw materials in parts by weight:
20-30 parts of glucose, 6-12 parts of peptone, 0.2-0.5 part of tremella polysaccharide, 8-12 parts of jerusalem artichoke powder, 1-3 parts of dipotassium hydrogen phosphate and 0.4-0.8 part of magnesium sulfate;
the preparation method of the anagen comprises the step (2) that the zymocyte consists of lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subspecies infantis.
2. The preparation containing metazoan according to claim 1, which comprises the following components in parts by weight:
64 parts of metazoan, 0.7 part of polyvinyl alcohol, 3 parts of absorption accelerator and 0.2 part of stabilizer.
3. The preparation containing metazoan according to claim 1, wherein the fermentation material comprises the following raw materials in parts by weight:
26 parts of glucose, 9 parts of peptone, 0.3 part of tremella polysaccharide, 10 parts of jerusalem artichoke powder, 2 parts of dipotassium hydrogen phosphate and 0.6 part of magnesium sulfate.
4. The formulation containing metazoa according to claim 1, wherein the fermentation tubes are made of lactobacillus casei, lactobacillus plantarum and bifidobacterium longum subsp. 6-8: 2-3.
5. The method for preparing a preparation containing metazoan according to any one of claims 1 to 4, comprising the steps of:
taking the latter biogen, adding polyvinyl alcohol at the stirring speed of 500r/min at 300-.
6. Use of a preparation containing metazoan according to any one of claims 1 to 4 for the preparation of a medicament against immunosenescence.
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| 高核心岩藻糖基化母乳促进子代肠道双歧杆菌和乳杆菌的优势生长;梅迪等;《中国微生态学杂志》;20200815(第08期);摘要 * |
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