CN102511716A - Glycyrrhiza polysaccharide granules and preparation method thereof - Google Patents
Glycyrrhiza polysaccharide granules and preparation method thereof Download PDFInfo
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- CN102511716A CN102511716A CN2011104509375A CN201110450937A CN102511716A CN 102511716 A CN102511716 A CN 102511716A CN 2011104509375 A CN2011104509375 A CN 2011104509375A CN 201110450937 A CN201110450937 A CN 201110450937A CN 102511716 A CN102511716 A CN 102511716A
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- radix glycyrrhizae
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Abstract
The invention discloses glycyrrhiza polysaccharide granules and a preparation method thereof; a tissue culture method is adopted to produce glycyrrhiza polysaccharide, and the glycyrrhiza polysaccharide granules can be prepared by the glycyrrhiza polysaccharide and other excipients. The glycyrrhiza polysaccharide granules comprise the components by parts: 100-150 parts of glycyrrhiza polysaccharide, 100-300 parts of sustained-release material, 100-200 parts of filler and 5-10 parts of adhesive. The glycyrrhiza polysaccharide is produced by being cultured by a fermentation tank; the developed glycyrrhiza polysaccharide granules are convenient to use and can be directly taken or taken with water; the produced glycyrrhiza polysaccharide has obvious immune dispensing activity and is capable of activating macrophage and tuberculosis (TB) lymphocyte, so that humoral immunity and cellular immunity can be enhanced, and the aims of improving the general immunity function and more effectively inhibiting tumours and acquired immunodeficiency syndrome (AIDS) can be achieved; furthermore, products developed by the polysaccharide have obvious health care function so as to have excellent develop space.
Description
Technical field
The present invention relates to a kind of health food and preparation method thereof, particularly a kind of Chinese medicinal granule health food.
Background technology
Glycyrrhiza (Glycyrrhiza) department of botany pulse family Papillionoideae herbaceos perennial, the research of plant cell suspension cultured are the important steps that Plant Tissue Breeding moves towards the industrialization and produces.Plant cell liquid suspension cultured to have an outstanding advantage cell proliferation rate fast, the culture of a large amount of uniformities can be provided and can carry out large-scale culture.Therefore; From the beginning of this century, utilize plant cell, the research that the method for tissue and organ culture is produced active medicinal matter has obtained breakthrough; Existing nearly 1000 kind of plant were carried out the research of cell cultivation aspect; According to estimates, from 400 various plants, set up tissue and cell culture, and therefrom isolated more than 600 kind of metabolite.Radix Glycyrrhizae is as a kind of traditional Chinese medicine; The good reputation of " ten sides, nine grass " is just arranged since ancient times, and the main active of Radix Glycyrrhizae has glycyrrhizic acid, Radix Glycyrrhizae brass and licorice polysaccharide, and the above two have had more deep research; And that the functional study of licorice polysaccharide mainly concentrates on is antitumor; Antiviral, regulate immunocompetence and remove the free radical aspect, seldom in the exploitation of licorice polysaccharide application facet.
Summary of the invention
In order to solve the problems of the prior art, the invention provides a kind of licorice polysaccharide granule and preparation method thereof, solved the problem of licorice polysaccharide in health products research and development field blank.
The present invention realizes through following technical scheme:
A kind of licorice polysaccharide granule adopts the method for tissue culture to produce licorice polysaccharide, is aided with other auxiliary material and processes the licorice polysaccharide granule, and its each composition proportion is following:
Described slow-release material is selected from one or more mixtures in HPMC or ethyl cellulose or cellulose acetate or the Aquacoat.
Described filler is selected from one or more mixtures in microcrystalline cellulose or sucrose or lactose or starch or sweet mellow wine or sorbierite or xylitol or dextrin or beta-schardinger dextrin-or calcium carbonate or calcium phosphate or the calcium monohydrogen phosphate.
Described adhesive is selected from the ethanolic solution of polyvinylpyrrolidone of ethanolic solution or 2-5% of the ethyl cellulose of 1-5%.
A kind of preparation method of licorice polysaccharide granule comprises the steps:
(1) extraction of licorice polysaccharide: the extracting liquorice cell powder, the distilled water of 15-25 times of weight of adding leaves standstill 5-12h, water-bath 1-5 time, each 1-3h.Repeat water-bath 2-5 time, merging filtrate is concentrated to certain volume, is 50-90% with absolute ethyl alcohol modulation final concentration, and 0-6 ℃ of hold over night removed supernatant, and with sevage method deproteinized, drying is pulverized, and obtains the licorice polysaccharide powder;
(2) extracting liquorice polysaccharide powder, filler, adhesive, water are granulated, and drying promptly gets the licorice polysaccharide granular preparation.
Said licorice polysaccharide is to obtain by extracting in the Radix Glycyrrhizae suspension cell, and its preparation method is:
(1) acquisition of Radix Glycyrrhizae aseptic seedling: select full grains, be bottle-green Radix Glycyrrhizae seed, the clorox with 5%～15% stirs sterilization to seed on magnetic stirring apparatus; Rotating speed is 200r/min～500r/min, and temperature is 20 ℃～30 ℃, sterilization time 20min～40min; MS contains 2%-5% sucrose in cultivating, and pH 5～6 before the sterilization is seeded in the triangular flask that contains MS solid medium 50ml; Be placed in 20～28 ℃ of illumination boxs, cultivated 30～35 days, obtain the Radix Glycyrrhizae aseptic seedling;
(2) acquisition of Radix Glycyrrhizae callus: the Radix Glycyrrhizae callus is to induce from Radix Glycyrrhizae aseptic seedling hypocotyl to get, and after repeatedly screening subculture, growth conditions is stable, and the callus subculture medium is the MS culture medium, adds 2,4-D (0.5～2mgL
-1), NAA (0.5～2mgL
-1), 6-BA (0.1～0.3mgL
-1), 2～5% sucrose, pH5 before the sterilization～6, subculture cycle 25～30 days;
(3) acquisition of Radix Glycyrrhizae suspension cell: select fast growth, loose, the flaxen Radix Glycyrrhizae callus of quality, transfer to after broken with the tweezers folder and carry out suspension cultured in the fluid nutrient medium; Cultivation and composition are the 3/4MS culture medium, add 2,4-D (0.5～2mgL
-1), NAA (0.5～2mgL
-1), 6-BA (0.1～0.3mgL
-1), 2～5% sucrose, cultivate 3～5 days for the first time after, the bulk callus that suspends is removed by filter, suspension cell is every at a distance from 15～20 days subcultures once, shaking speed 110～120rmin
-1, cultivation temperature (20～28) ℃, through the screening of going down to posterity for several times, the clone of final obtained performance excellence;
(4) Radix Glycyrrhizae suspension cell bubbling style reactor is cultivated: utilize the 5L bubbling style reactor of design voluntarily; Inoculum concentration is 5%～10% fresh weight licorice cell, and the fermentation tank throughput is 200L/min～500L/min, and cultivation and composition are the 3/4MS culture medium; Add 2,4-D (0.5～2mgL
-1), NAA (0.5～2mgL
-1), 6-BA (0.1～0.3mgL
-1), 2～5% sucrose, pH value are 5～9, cultivation cycle is 15～20 days.
Advantage of the present invention is advantage: the present invention utilizes the training method of fermentation tank culture to produce licorice polysaccharide, develops the licorice polysaccharide granule, and is easy to use; Can directly swallow, also can wash by water and take, produce licorice polysaccharide and have tangible immunity regulatin remedy activity; Its ability activating macrophage; The TB lymphocyte strengthens humoral immunity and cellular immunity, improves the general immunity function and more effectively suppresses the purpose of tumour and AIDS; Utilize this polysaccharide to develop product and have obvious health care, have development space.
The packing specification of health food particle provided by the invention is: the 10g/ bag is diluted in the 200ml hot water.Instructions about how to take medicine: sooner or later respectively obey one bag every day.
The specific embodiment
Below in conjunction with specific embodiment the present invention is further described.
The preparation of licorice polysaccharide
(1) acquisition of Radix Glycyrrhizae aseptic seedling: select full grains, be bottle-green Radix Glycyrrhizae seed, the clorox with 10% stirs sterilization to seed on magnetic stirring apparatus; Rotating speed is 300r/min, and temperature is 25 ℃, sterilization time 30min; MS contains 3% sucrose in cultivating, and pH5.9 before the sterilization is seeded in the triangular flask that contains MS solid medium 50ml; Be placed in 25 ℃ of illumination boxs, cultivated 30 days, obtain the Radix Glycyrrhizae aseptic seedling.
(2) acquisition of Radix Glycyrrhizae callus: the Radix Glycyrrhizae callus is to induce from Radix Glycyrrhizae aseptic seedling hypocotyl to get, and after repeatedly screening subculture, growth conditions is stable.The callus subculture medium is the MS culture medium, adds 2,4-D (1mgL
-1), NAA (1mgL
-1), 6-BA (0.2mgL
-1), 3% sucrose, pH5.9 before the sterilization, subculture cycle 28 days.
(3) acquisition of Radix Glycyrrhizae suspension cell: select fast growth, loose, the flaxen Radix Glycyrrhizae callus of quality, transfer to after broken with the tweezers folder and carry out suspension cultured in the fluid nutrient medium.Cultivation and composition are the 3/4MS culture medium, add 2,4-D (1mgL
-1), NAA (1mgL
-1), 6-BA (0.2mgL
-1), 3% sucrose, cultivate 3 days for the first time after, the bulk callus that suspends is removed by filter, suspension cell is every at a distance from 18 days subcultures once.Shaking speed 110～120rmin
-1, cultivation temperature (20～28) ℃.Through the screening of going down to posterity for several times, the excellent clone of final obtained performance.
(4) Radix Glycyrrhizae suspension cell bubbling style reactor is cultivated: utilize the 5L bubbling style reactor of design voluntarily, inoculum concentration is 8% fresh weight licorice cell, and the fermentation tank throughput is 300L//min, and cultivation and composition are the 3/4MS culture medium, adds 2,4-D (1mgL
-1), NAA (1mgL
-1), 6-BA (0.2mgL
-1), 3% sucrose, pH value are 5.9, cultivation cycle is 18 days.
Embodiment 1:
With the Radix Glycyrrhizae suspension cell is raw material, gets cell powder 2kg, adds the water of 20 times of licorice cell powder qualities, soaks 12h; Heated water bath is extracted, and 100 ℃ of temperature are extracted 1h, repeat to extract 2 times; Be concentrated into certain volume, add 4 times of volume absolute ethyl alcohols, under 4 ℃ of environment, leave standstill 12h, remove supernatant; Remove deproteinized with the sevage method, 45 ℃ of forced air drying 24h grind, and obtain the licorice polysaccharide powder.
Get the above-mentioned licorice polysaccharide of 100g, the 200g HPMC, the 150g lactose after mixing, is crossed 100 mesh sieves, adds the ethanolic solution of the ethyl cellulose of 20g1-5%, and 60 ℃ of dry 5h cross the whole grain of 20 mesh then, promptly obtain the licorice polysaccharide granular preparation.
Embodiment 2:
With the Radix Glycyrrhizae suspension cell is raw material, gets cell powder 2kg, adds the water of 15 times of licorice cell powder qualities, soaks 5h; Heated water bath is extracted, and 100 ℃ of temperature are extracted 3h, repeat to extract 1 time; Be concentrated into certain volume, add 4 times of volume absolute ethyl alcohols, under 4 ℃ of environment, leave standstill 12h, remove supernatant; Remove deproteinized with the sevage method, 45 ℃ of forced air drying 24h grind, and obtain white licorice polysaccharide powder.
Get the above-mentioned licorice polysaccharide of 100g, the 200g ethyl cellulose, 150g sweet mellow wine after mixing, is crossed 100 mesh sieves, adds the ethanolic solution of the ethyl cellulose of 20g1-5%, and 60 ℃ of dry 5h cross the whole grain of 20 mesh then, promptly obtain the licorice polysaccharide granular preparation.
Embodiment 3:
With the Radix Glycyrrhizae suspension cell is raw material, gets cell powder 2kg, adds the water of 25 times of licorice cell powder qualities, soaks 8h; Heated water bath is extracted, and 100 ℃ of temperature are extracted 1h, repeat to extract 5 times; Be concentrated into certain volume, add 4 times of volume absolute ethyl alcohols, under 4 ℃ of environment, leave standstill 12h, remove supernatant; Remove deproteinized with the sevage method, 45 ℃ of forced air drying 24h grind, and obtain white licorice polysaccharide powder.
Get the above-mentioned licorice polysaccharide of 100g, the 200g cellulose acetate, the 150g sorbierite after mixing, is crossed 100 mesh sieves, adds the ethanolic solution of the ethyl cellulose of 20g1-5%, and 60 ℃ of dry 5h cross the whole grain of 20 mesh then, promptly obtain the licorice polysaccharide granular preparation.
Embodiment 4:
With the Radix Glycyrrhizae suspension cell is raw material, gets cell powder 2kg, adds the water of 20 times of licorice cell powder qualities, soaks 12h; Heated water bath is extracted, and 100 ℃ of temperature are extracted 1.5h, repeat to extract 2 times; Be concentrated into certain volume, add 4 times of volume absolute ethyl alcohols, under 4 ℃ of environment, leave standstill 12h, remove supernatant; Remove deproteinized with the sevage method, 45 ℃ of forced air drying 24h grind, and obtain white licorice polysaccharide powder.
Get the above-mentioned licorice polysaccharide of 100g, 200g Aquacoat, 150g xylitol; After mixing, cross 100 mesh sieves, add the ethanolic solution of the ethyl cellulose of 20g1-5%; 60 ℃ of dry 5h cross the whole grain of 20 mesh then, promptly obtain the licorice polysaccharide granular preparation.
Embodiment 5:
With the Radix Glycyrrhizae suspension cell is raw material, gets cell powder 2kg, adds the water of 20 times of licorice cell powder qualities, soaks 12h; Heated water bath is extracted, and 100 ℃ of temperature are extracted 1h, repeat to extract 2 times; Be concentrated into certain volume, add 4 times of volume absolute ethyl alcohols, under 4 ℃ of environment, leave standstill 12h, remove supernatant; Remove deproteinized with the sevage method, 45 ℃ of forced air drying 24h grind, and obtain white licorice polysaccharide powder.
Get the above-mentioned licorice polysaccharide of 100g, 200g slow-release material (wherein HPMC 100g, ethyl cellulose 100g); 150g filler (wherein lactose 100g, sucrose 50g) is after mixing; Cross 100 mesh sieves, add the ethanolic solution of the ethyl cellulose of 20g1-5%, 60 ℃ of dry 5h; Cross the whole grain of 20 mesh then, promptly obtain the licorice polysaccharide granular preparation.
Test Example:
Animal experiment by University Of Tianjin modern Chinese herbal medicine quality research development centre is done by " health food function assessment assessment process and method of inspection method " immunological regulation of the present invention, oxidation-resisting health-care food granule is reported as follows:
1. every day Kunming white mouse oral administration is invented basic, normal, high three dose groups that particle is equivalent to 5 times, 10 times and 30 times recommended amounts of people's recommended amounts respectively; A control group is set in addition; Feeding is 30 days altogether, carries out carbon clearance test and dinitrofluorobenzene inducing mouse DTH test, and the result obtains: 1. clean up ability with the carbon of the index bidding documents mouse of swallowing; Through statistical analysis, the index of swallowing of three dose groups is significantly higher than the index of swallowing of control group.2. represent the degree of DHT with the difference of mouse left and right sides ear weight, through statistical analysis, the difference of dose groups is significantly higher than the difference of control group.Above-mentioned animal test results shows that health care particle of the present invention has tangible ability of regulating immunity.
2. every day the Drosophila melanogaster oral administration is invented basic, normal, high three dose groups that particle is equivalent to 5 times, 10 times and 30 times recommended amounts of people's recommended amounts respectively, an aged group of contrast is set in addition, stop to life span of drosophila melanogaster.Every day 18 months rats are given basic, normal, high three dose groups of 5 times, 10 times and the 30 times recommended amounts that particle of the present invention is equivalent to people's recommended amounts respectively; Design a control group in addition; Totally 45 days; The result shows: 1. dose groups and control group compare, and the half death, average life span and the maximum life span that are tried fruit bat are through statistical analysis, and difference has conspicuousness (P＜0.05).2. dose groups compares with the aged group of contrast, and the SOD in the rat blood serum (superoxide dismutase) vigor raises through statistical analysis, and difference has conspicuousness (P＜0.05).Above-mentioned animal experiment proof particle of the present invention has and obviously has non-oxidizability.
Claims (6)
1. a licorice polysaccharide granule is characterized in that, adopts the method for tissue culture to produce licorice polysaccharide, is aided with other auxiliary material and processes the licorice polysaccharide granule, and its each composition proportion is following:
2. licorice polysaccharide granule according to claim 1 is characterized in that, described slow-release material is selected from one or more mixtures in HPMC or ethyl cellulose or cellulose acetate or the Aquacoat.
3. licorice polysaccharide granule according to claim 1; It is characterized in that described filler is selected from one or more mixtures in microcrystalline cellulose or sucrose or lactose or starch or sweet mellow wine or sorbierite or xylitol or dextrin or beta-schardinger dextrin-or calcium carbonate or calcium phosphate or the calcium monohydrogen phosphate.
4. licorice polysaccharide granule according to claim 1 is characterized in that, described adhesive is selected from the ethanolic solution of polyvinylpyrrolidone of ethanolic solution or 2-5% of the ethyl cellulose of 1-5%.
5. the preparation method of a licorice polysaccharide granule is characterized in that, comprises the steps:
(1) extraction of licorice polysaccharide: the extracting liquorice cell powder, the distilled water of 15-25 times of weight of adding leaves standstill 5-12h, water-bath 1-5 time, each 1-3h.Repeat water-bath 2-5 time, merging filtrate is concentrated to certain volume, is 50-90% with absolute ethyl alcohol modulation final concentration, and 0-6 ℃ of hold over night removed supernatant, and with sevage method deproteinized, drying is pulverized, and obtains the licorice polysaccharide powder;
(2) extracting liquorice polysaccharide powder, filler, adhesive, water are granulated, and drying promptly gets the licorice polysaccharide granular preparation.
6. according to the preparation method of the said licorice polysaccharide granule of claim 5, it is characterized in that said licorice polysaccharide is to obtain by extracting in the Radix Glycyrrhizae suspension cell, its preparation method is:
(1) acquisition of Radix Glycyrrhizae aseptic seedling: select full grains, be bottle-green Radix Glycyrrhizae seed, the clorox with 5%～15% stirs sterilization to seed on magnetic stirring apparatus; Rotating speed is 200r/min～500r/min, and temperature is 20 ℃～30 ℃, sterilization time 20min～40min; MS contains 2%-5% sucrose in cultivating, and pH 5～6 before the sterilization is seeded in the triangular flask that contains MS solid medium 50ml; Be placed in 20～28 ℃ of illumination boxs, cultivated 30～35 days, obtain the Radix Glycyrrhizae aseptic seedling;
(2) acquisition of Radix Glycyrrhizae callus: the Radix Glycyrrhizae callus is to induce from Radix Glycyrrhizae aseptic seedling hypocotyl to get, and after repeatedly screening subculture, growth conditions is stable, and the callus subculture medium is the MS culture medium, adds 2,4-D (0.5～2mgL
-1), NAA (0.5～2mgL
-1), 6-BA (0.1～0.3mgL
-1), 2～5% sucrose, pH5 before the sterilization～6, subculture cycle 25～30 days;
(3) acquisition of Radix Glycyrrhizae suspension cell: select fast growth, loose, the flaxen Radix Glycyrrhizae callus of quality, transfer to after broken with the tweezers folder and carry out suspension cultured in the fluid nutrient medium; Cultivation and composition are the 3/4MS culture medium, add 2,4-D (0.5～2mgL
-1), NAA (0.5～2mgL
-1), 6-BA (0.1～0.3mgL
-1), 2～5% sucrose, cultivate 3～5 days for the first time after, the bulk callus that suspends is removed by filter, suspension cell is every at a distance from 15～20 days subcultures once, shaking speed 110～120rmin
-1, cultivation temperature (20～28) ℃, through the screening of going down to posterity for several times, the clone of final obtained performance excellence;
(4) Radix Glycyrrhizae suspension cell bubbling style reactor is cultivated: utilize the 5L bubbling style reactor of design voluntarily; Inoculum concentration is 5%～10% fresh weight licorice cell, and the fermentation tank throughput is 200L/min～500L/min, and cultivation and composition are the 3/4MS culture medium; Add 2,4-D (0.5～2mgL
-1), NAA (0.5～2mgL
-1), 6-BA (0.1～0.3mgL
-1), 2～5% sucrose, pH value are 5～9, cultivation cycle is 15～20 days.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105878188A (en) * | 2014-11-06 | 2016-08-24 | 王辉 | Fermented cordyceps sinensis extracellular polysaccharide granules and preparation method thereof |
| CN109156815A (en) * | 2018-09-17 | 2019-01-08 | 嘉应学院 | A kind of kudzu cultivating ganoderma mushroom bran granule and preparation method thereof |
| CN111718888A (en) * | 2020-06-23 | 2020-09-29 | 中国医学科学院药用植物研究所 | Culture method for improving glycyrrhizic acid content in suspension culture cells of liquorice |
| CN115634233A (en) * | 2022-11-09 | 2023-01-24 | 陕西师范大学 | Application of glycyrrhiza polysaccharide in preparation of product for relieving triptolide toxicity |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1138787C (en) * | 2000-09-20 | 2004-02-18 | 天津市贝特科技发展有限公司 | Natural plant polyose |
| CN1234368C (en) * | 2003-08-01 | 2006-01-04 | 天津市贝特科技发展有限公司 | New medicinal use of liquorice polysaccharide and its medicinal preparation |
| CN101766680A (en) * | 2008-12-31 | 2010-07-07 | 李志方 | Comprehensive extraction method of glycyrrhiza |
-
2011
- 2011-12-29 CN CN2011104509375A patent/CN102511716A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1138787C (en) * | 2000-09-20 | 2004-02-18 | 天津市贝特科技发展有限公司 | Natural plant polyose |
| CN1234368C (en) * | 2003-08-01 | 2006-01-04 | 天津市贝特科技发展有限公司 | New medicinal use of liquorice polysaccharide and its medicinal preparation |
| CN101766680A (en) * | 2008-12-31 | 2010-07-07 | 李志方 | Comprehensive extraction method of glycyrrhiza |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105878188A (en) * | 2014-11-06 | 2016-08-24 | 王辉 | Fermented cordyceps sinensis extracellular polysaccharide granules and preparation method thereof |
| CN109156815A (en) * | 2018-09-17 | 2019-01-08 | 嘉应学院 | A kind of kudzu cultivating ganoderma mushroom bran granule and preparation method thereof |
| CN111718888A (en) * | 2020-06-23 | 2020-09-29 | 中国医学科学院药用植物研究所 | Culture method for improving glycyrrhizic acid content in suspension culture cells of liquorice |
| CN111718888B (en) * | 2020-06-23 | 2022-08-19 | 中国医学科学院药用植物研究所 | Culture method for improving glycyrrhizic acid content in suspension culture cells of liquorice |
| CN115634233A (en) * | 2022-11-09 | 2023-01-24 | 陕西师范大学 | Application of glycyrrhiza polysaccharide in preparation of product for relieving triptolide toxicity |
| CN115634233B (en) * | 2022-11-09 | 2024-05-14 | 陕西师范大学 | Application of licorice polysaccharide in preparation of triptolide toxicity relieving product |
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Application publication date: 20120627 |