CN1297569C - Preparation method of ganoderma polysaccharide - Google Patents
Preparation method of ganoderma polysaccharide Download PDFInfo
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- CN1297569C CN1297569C CNB2004100673012A CN200410067301A CN1297569C CN 1297569 C CN1297569 C CN 1297569C CN B2004100673012 A CNB2004100673012 A CN B2004100673012A CN 200410067301 A CN200410067301 A CN 200410067301A CN 1297569 C CN1297569 C CN 1297569C
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Abstract
The present invention relates to a method for preparing macro-molecular ganoderma polysaccharide, which comprises four steps: rough extraction of ganoderma, rough filtration, ultrafiltration and drying. The preparing method can be utilized to obtain macro-molecular ganoderma polysaccharide with a molecular weight larger than 500000 and a polysaccharide content higher than 50%. Besides, the anti-tumor activity and the immunity are enhanced.
Description
Technical field
The present invention relates to a kind of preparation method of ganoderma polysaccharide, relate in particular to a kind of preparation method of glossy ganoderma macromolecular polysaccharide.
Background technology
The separation and Extraction water-soluble polysaccharide generally adopts the ethanol step-by-step precipitation method at present from glossy ganoderma, be easy to scale operation, its cardinal principle is to make the polysaccharide dehydration by the specific inductivity that reduces the aqueous solution, thereby produce the purpose that precipitation reaches separating polyose, be applicable to most of water-soluble polysaccharides, but the polysaccharide impurity that obtains is many, and active constituent content is low, and specificity is not high.Fractional precipitation comes purifying as further employing Different concentrations of alcohol, and selectivity is relatively poor, and the rate of recovery is low, obtains polysaccharide content about 30%, and is the mixture of polysaccharide, will use a large amount of organic solvents simultaneously, has improved cost.In addition, some breadboard researchs are arranged also, aqueous extract obtains ganoderan after removing albumen and dialysis, but complex process needs organic solvents such as adding chloroform and propyl carbinol, is difficult for becoming suitability for industrialized production.
The research report that the immunoregulatory effect of glossy ganoderma macromolecular polysaccharide composition is existing a large amount of, and can reach antitumous effect by improving body's immunological function, be the effective constituent of antitumous effect.But often contain multiple polysaccharide component in a kind of plant or the fungi, the size of molecular weight also has nothing in common with each other, its activity also has nothing in common with each other, the polysaccharide component that also contains different molecular weight sections in the glossy ganoderma, for improving the effect of polysaccharide, non-activity or active not high composition must be removed, be enriched with active polysaccharide part.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of preparation method of glossy ganoderma macromolecular polysaccharide, the shortcoming that the purity of polysaccharide of extracting in the prior art is low to overcome, specificity is not high, be difficult to suitability for industrialized production.
Principle of the present invention is: activated polysaccharide in the Ganoderma extract is partly come the principle of enrichment with suitable ultra-filtration membrane, removing does not have active polysaccharide component.
The preparation method of a kind of glossy ganoderma macromolecular polysaccharide that provides of the present invention comprises the steps:
1. slightly carrying of glossy ganoderma: with the Ganoderma sporophore is raw material, is ground into coarse grain, add in the water of 5-15 times of weight, and soak at room temperature 10-60 minute, be heated to 60-100 ℃, kept this temperature 60-120 minute, filter.Filter residue repeats above-mentioned steps, extracts 1-2 time merging filtrate again.
2. the coarse filtration of glossy ganoderma crude extract: filtrate is used 100 microns membrane filtration again with 50-200 purpose filter screen coarse filtration.
3. glossy ganoderma filtrate ultrafiltration: use ultra-filtration membrane to carry out ultrafiltration, the molecular weight cutoff value of ultra-filtration membrane is 500,000 dalton, and pressure is lower than 0.3Mpa, and filtrate is divided into two parts, greater than the part of 500,000 molecular weight with less than the part of 500,000 molecular weight.
4. dry: the part convection drying greater than 500,000 molecular weight promptly obtains required macromolecular polysaccharide.
Can obtain steady quality, effective component content height and molecular-weight average by above-mentioned preparation method is strictly controlled at greater than 500,000 daltonian ganoderan compositions, present method has not only kept the anti-tumor activity of polysaccharide, and the content of ganoderan is improved, and polysaccharide content surpasses 50%.So this preparation method be a kind of easy, quick, efficient, cost is low, pollution-free, suitable industrial method that purity of polysaccharide is high.
The biological activity assay of the macromole ganoderan of preparation gained:
According to the Stanley reported method, select the Balb/c mouse of 8～10 ages in week, body weight (28 ± 1) g, put to death, extract the marrow in mouse thigh, the focile, be incubated at after the dispersion in the complete DMEM substratum that contains 10%L929 cells and supernatant (behind the L929 cell strain cultivation 3d, removing the substratum of cell).Behind the 3d, remove adherent cell, non-adherent cell is placed the complete DMEM substratum that adds the 10%L929 cells and supernatant.After continuing to cultivate 3d, remove non-adherent cell, collect adherent cell, be the scavenger cell in mouse bone marrow cells source.The mouse macrophage activity ratio is measured 1 * 10
5The scavenger cell suspension of individual/mL adds in 96 orifice plates, and every hole adds 180 μ L, and adds sample 20 μ L.Under 37 ℃, 5% CO2 condition, cultivated 3 days, add 20 μ L Alamar Blue reagent, cultivate 6hr again, read the absorbancy that the plate instrument is measured 570nm and 600nm place automatically with ELISA, the formula according to Alamar Blue reagent calculates the activity ratio of various samples to scavenger cell then.Calculation formula is: macrophage activation rate (%)=[80856 * A
λ 570(sample)-11726 * A
λ 600(sample)]/[80856 * A
λ 570(contrast)-11726 * A
λ 600(contrast)] * 100%.
Molecular weight greater than 500,000 parts to the mouse macrophage activity ratio greater than 200%, can obviously strengthen the phagocytic activity of scavenger cell, thus the anti-tumor capacity of enhancing body.
The detection of polysaccharide content:
With phenol-vitriol oil method, precision takes by weighing this product 0.5g and puts in the 100ml beaker, adds the about 80ml of water, boils and extracts 1 hour postcooling to room temperature, adds in the 100ml volumetric flask.Be diluted with water to scale, shake up, 25 times of redilution.The accurate sample 1.0ml that draws puts in the 15ml test tube, adds phenolsulfuric acid reagent respectively, measures optical density A at the 490nm place.With the dextran is that standard substance calculate sugar contents, and measures reducing sugar content with DNS, and the content of polysaccharide equals total reducing sugar and subtracts reducing sugar.Molecular weight greater than the polysaccharide content of 500,000 parts greater than more than 50%.
Embodiment:
Embodiment 1:
1. slightly carrying of glossy ganoderma: with the Ganoderma sporophore is raw material, is ground into coarse grain, adds in the water of 5 times of weight, and soak at room temperature 60 minutes is heated to 60 ℃, keeps this temperature 60 minutes, filters.Filter residue repeats above-mentioned steps, extracts once merging filtrate again.
2. the coarse filtration of glossy ganoderma crude extract: filtrate is used 100 microns membrane filtration again with 100 purpose filter screen coarse filtration.
3. glossy ganoderma filtrate ultrafiltration: use ultra-filtration membrane to carry out ultrafiltration, the molecular weight cutoff value of ultra-filtration membrane is 500,000 dalton, and pressure is lower than 0.3Mpa, and filtrate is divided into two parts, greater than the part of 500,000 molecular weight with less than the part of 500,000 molecular weight.
4. dry: the part spraying drying greater than 500,000 molecular weight promptly obtains required polysaccharide.
The polysaccharide of preparation is 320% to the mouse macrophage activity ratio.Polysaccharide content is 55%.
Embodiment 2:
1. slightly carrying of glossy ganoderma: with the Ganoderma sporophore is raw material, is ground into coarse grain, adds in the water of 10 times of weight, and soak at room temperature 30 minutes is heated to 70 ℃, keeps this temperature 60 minutes, filters.Filter residue repeats above-mentioned steps, extracts once merging filtrate again.
2. the coarse filtration of glossy ganoderma crude extract: filtrate is used 100 microns membrane filtration again with 200 purpose filter screen coarse filtration.
3. glossy ganoderma filtrate ultrafiltration: use ultra-filtration membrane to carry out ultrafiltration, the molecular weight cutoff value of ultra-filtration membrane is 500,000 dalton, and pressure is lower than 0.3Mpa, and filtrate is divided into two parts, greater than the part of 500,000 molecular weight with less than the part of 500,000 molecular weight.
4. dry: as, promptly to obtain required polysaccharide greater than the direct lyophilize of the part of 500,000 molecular weight.
The polysaccharide of preparation is 290% to the mouse macrophage activity ratio.Polysaccharide content is 53%.
Claims (1)
1. a preparation method of ganoderma polysaccharide is characterized in that this method comprises the steps:
1. slightly carrying of glossy ganoderma: with the Ganoderma sporophore is raw material, is ground into coarse grain, add in the water of 5-15 times of weight, and soak at room temperature 10-60 minute, be heated to 60-100 ℃, kept this temperature 60-120 minute, filter; Filter residue repeats above-mentioned steps, extracts 1-2 time merging filtrate again;
2. the coarse filtration of glossy ganoderma crude extract: filtrate is used 100 microns membrane filtration again with 50-200 purpose filter screen coarse filtration;
3. glossy ganoderma filtrate ultrafiltration: use ultra-filtration membrane to carry out ultrafiltration, the molecular weight cutoff value of ultra-filtration membrane is 500,000, and pressure is lower than 0.3Mpa, and filtrate is divided into two parts, greater than the part of 500,000 molecular weight with less than the part of 500,000 molecular weight;
4. dry: the part convection drying greater than 500,000 molecular weight promptly obtains required macromolecular polysaccharide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100673012A CN1297569C (en) | 2004-10-20 | 2004-10-20 | Preparation method of ganoderma polysaccharide |
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2004100673012A CN1297569C (en) | 2004-10-20 | 2004-10-20 | Preparation method of ganoderma polysaccharide |
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| CN1618819A CN1618819A (en) | 2005-05-25 |
| CN1297569C true CN1297569C (en) | 2007-01-31 |
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| CNB2004100673012A Expired - Fee Related CN1297569C (en) | 2004-10-20 | 2004-10-20 | Preparation method of ganoderma polysaccharide |
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Families Citing this family (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100409856C (en) * | 2006-09-13 | 2008-08-13 | 贵州省科晖制药厂 | Glossy ganoderma extractum preparing process |
| CN101434660B (en) * | 2008-08-01 | 2011-08-17 | 湖北省农业科学院农产品加工与核农技术研究所 | Method for extracting Ganoderma lucidum polysaccharide by ultrasonic wave-microwave combination |
| CN101508739B (en) * | 2009-03-24 | 2011-02-16 | 湖北省农业科学院农产品加工与核农技术研究所 | Radio frequency auxiliary method for extracting glossy ganoderma seed polysaccharide |
| CN102040748B (en) * | 2009-10-16 | 2013-04-24 | 上海医药工业研究院 | Ganoderma sinensis mycelium anti-tumor polysaccharide component GS-C as well as preparation method and application thereof |
| CN101935358A (en) * | 2010-09-10 | 2011-01-05 | 上海市农业科学院 | Ganoderma lucidum polysaccharide and preparation method thereof |
| CN102120779B (en) * | 2011-04-18 | 2012-07-25 | 夏历 | Efficient extraction technology of ganodorma lucidum fermention mycelium homogeneous polysaccharides |
| CN102585027B (en) * | 2012-02-23 | 2014-06-11 | 上海市农业科学院 | Coprinus-comatus macromolecular polysaccharide and preparation method thereof |
| CN102718880A (en) * | 2012-06-23 | 2012-10-10 | 上海市农业科学院 | Ganoderma lucidum polysaccharide of high molecular weight and detection method thereof |
| CN104059162B (en) * | 2014-06-25 | 2016-03-23 | 上海市农业科学院 | A kind of ganoderan and preparation method thereof |
| TWI543765B (en) * | 2015-05-25 | 2016-08-01 | 長庚生物科技股份有限公司 | Method to prepare ganoderma lucidum polysaccharides possessing anti-obesity properties |
| CN104945528A (en) * | 2015-07-08 | 2015-09-30 | 广州白云山汉方现代药业有限公司 | Ganoderma lucidum spores polysaccharides preparing method |
| CN106511403B (en) * | 2017-01-09 | 2019-07-12 | 仙芝科技(福建)股份有限公司 | A kind of ganoderma lucidum tablet and preparation method thereof |
| CN113768141B (en) * | 2021-09-18 | 2022-10-18 | 广东粤微食用菌技术有限公司 | Anti-oxidation repairing ganoderma lucidum extract and preparation method thereof |
| CN115785300A (en) * | 2022-11-30 | 2023-03-14 | 上海市农业科学院 | A Ganoderma water extraction ultrafiltrate rich in beta- (1 → 3,1 → 6) -dextran, and its preparation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6372629A (en) * | 1986-09-17 | 1988-04-02 | Kyoritsu Yakuhin Kogyo Kk | Production of physiologically active extract from ganoderma lucidum karst. |
| CN1422872A (en) * | 2001-11-26 | 2003-06-11 | 刘年生 | High-germanium ganoderic polysaccharide and ganoderic acid extraction method |
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2004
- 2004-10-20 CN CNB2004100673012A patent/CN1297569C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6372629A (en) * | 1986-09-17 | 1988-04-02 | Kyoritsu Yakuhin Kogyo Kk | Production of physiologically active extract from ganoderma lucidum karst. |
| CN1422872A (en) * | 2001-11-26 | 2003-06-11 | 刘年生 | High-germanium ganoderic polysaccharide and ganoderic acid extraction method |
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| CN1618819A (en) | 2005-05-25 |
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