CN102653567A - Extracting process of cordyceps sinensis mycelium polysaccharide of composite preparation for preventing tumor - Google Patents
Extracting process of cordyceps sinensis mycelium polysaccharide of composite preparation for preventing tumor Download PDFInfo
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- CN102653567A CN102653567A CN2012101134220A CN201210113422A CN102653567A CN 102653567 A CN102653567 A CN 102653567A CN 2012101134220 A CN2012101134220 A CN 2012101134220A CN 201210113422 A CN201210113422 A CN 201210113422A CN 102653567 A CN102653567 A CN 102653567A
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Abstract
The invention discloses an extracting process of cordyceps sinensis mycelium polysaccharide of a composite preparation for preventing a tumor, and the method comprises the following steps of: boiling 100 g of mycelium powder obtained by concentrating a cordyceps sinensis fermentation extracting solution; heating a filtering solution to obtain a concentrated solution; collecting the concentrated solution in a water bath so as to be concentrated; packaging the concentrated solution into eight centrifugal tubes so as to be centrifuged to obtain a water-soluble cordyceps sinensis polysaccharide solution; collecting liquid supernatant in the solution and slowly adding the liquid supernatant into absolute ethyl alcohol which is 4 times as much as the volume of the liquid supernatant; standing to obtain alcohol sediment; centrifuging and collecting alcohol precipitated polysaccharide in each centrifugal tube into each centrifugal tube; washing the alcohol precipitated polysaccharide twice by use of ethanol and freezing and drying in vacuum and at a low temperature to obtain a rough cordyceps sinensis mycelium polysaccharide; dissolving the rough cordyceps sinensis mycelium polysaccharide into distilled water and re-treating by suitable amount of the ethanol to obtain sediment; and freezing and drying the sediment in vacuum and at the low temperature to obtain cordyceps sinensis mycelium polysaccharide powder. The extracting process disclosed by the invention is used for finely sieving components in the rough polysaccharide to sieve components for accelerating anti-tumor immune cells to be multiplied, so that the anti-tumor immune effect of an organism is enhanced.
Description
Technical field
The present invention relates to the prophylaxis of tumours compound formulation, be specifically related to a kind of Cordyceps mycelium polysaccharide extracting process that is used for the prophylaxis of tumours compound formulation.
Background technology
Occurred frequently in view of tumour, and prevent, treatment means is limited, and anti-oxidant importance in prophylaxis of tumours presses for a kind of compound preparation in the market, plays prophylaxis of tumours and oxidation resistant composite efficacy.
Lack at present preventing cancer and oxidation resistant product, and the theory of institute's foundation is comparatively single, lacks the mutual checking of traditional Chinese and western medicine: or excavate traditional prescription according to theory of traditional Chinese medical science, lack the theory and the experimental evidence support of modern medicine; Be difficult to satisfy prophylaxis of tumours and oxidation resistant demand simultaneously.
The existing Cordyceps mycelium polysaccharide extracting process that is used for the prophylaxis of tumours compound formulation is a Crude polysaccharides, and the composition in the Crude polysaccharides segments, and what have has an immunostimulation; What have has an immunoregulation effect; The use that mixes can produce effect and offset, and the present invention segments the composition of entomophyte Crude polysaccharides; One-component to promote tumour immunity is a target, more remarkable effect.
Summary of the invention
The present invention provides a kind of Cordyceps mycelium polysaccharide extracting process that is used for the prophylaxis of tumours compound formulation; This Cordyceps mycelium polysaccharide extracting process; Composition in the Crude polysaccharides in the prior art segments; Filter out component, strengthened antitumor immunity of organism power effect with the cell proliferation of promotion antineoplastic immune.
For achieving the above object, the technical scheme that the present invention adopts is: a kind of Cordyceps mycelium polysaccharide extracting process that is used for the prophylaxis of tumours compound formulation may further comprise the steps:
Step 1, get the Chinese caterpillar fungus fermentation extracting solution and concentrate the bacterium powder 100g form, add 4L H
2O boils behind the 4h while hot filtered through gauze and removes residue;
Step 2, the filtrating heating that will obtain through step 1 be concentrated into 1L and obtain liquid concentrator;
Step 3, the liquid concentrator of step 2 is collected in the 1L beaker, 70 ℃ of water-baths are concentrated into 400ml;
Step 4, liquid concentrator branch in the step 3 is filled in the centrifuge tube of 8 50ml, under 4500 rpm centrifugation rates, handles 10min, obtain water-soluble Cordyceps polysaccharide solution;
Step 5, collect the about 400ml of supernatant in step 4 solution, the removal insoluble impurities to the 2L beaker, leaves standstill the acquisition precipitation solution after slowly adding 4 times of volume absolute ethyl alcohols;
Step 6, the precipitation solution branch of step 5 is filled in 40 50ml centrifuge tubes, alcohol precipitation polysaccharide in said each centrifuge tube is collected in 4 50ml centrifuge tubes after under 4500 rpm centrifugation rates, handling 10min;
Step 7, wash through the said alcohol precipitation polysaccharide of step 62 times through 100% ethanol, the vacuum and low temperature lyophilize obtains the Cordyceps mycelium polysaccharide;
Step 8, the Cordyceps mycelium polysaccharide of step 7 is dissolved in the zero(ppm) water; Stir after 2 hours; The 10000rpm centrifugation rate is handled down and was obtained supernatant in 5 minutes, in this supernatant, adds 100% ethanol, and the limit edged stirs; After 4 ℃ of depositions hour, under the 10000rpm centrifugation rate, handle again 5 minutes post precipitations at this moment throw out obtain said Cordyceps mycelium polysaccharide powder through the vacuum and low temperature lyophilize.
Further improvement project in the technique scheme is following:
1, in the such scheme, carries out stirring operation when filtrating is heated in the said step 2 simultaneously.
2, in the such scheme, in the said step 2 in 70 ℃ of water-baths the hold-time be 24h.
3, in the such scheme, dwell temperature is 4 ℃ in the said step 5, and the time is 12h.
Because the technique scheme utilization, the present invention compared with prior art has advantage and effect:
The present invention is used for the Cordyceps mycelium polysaccharide powder extraction process of prophylaxis of tumours compound formulation; Composition in the Crude polysaccharides segments; Adopt the multistep extraction step, obtain various ingredients, and compare the physiological function of different components; One-component to promote tumour immunity is a target, makes the product of acquisition more remarkable at enhancing antineoplastic immune aspect of performance.
Description of drawings
Accompanying drawing 1 is the present invention and the prior art comparison diagram at the killing tumor cell aspect of performance.
Embodiment
Below in conjunction with accompanying drawing and embodiment the present invention is further described:
Embodiment: a kind of Cordyceps mycelium polysaccharide extracting process that is used for the prophylaxis of tumours compound formulation may further comprise the steps:
Step 1, get the Chinese caterpillar fungus fermentation extracting solution and concentrate the bacterium powder 100g form, this bacterium powder is formed by the Chinese caterpillar fungus fermentation fluid dewatering, adding 4L H
2O boils 4h, and filtered through gauze is removed residue while hot.Repeat 3 times;
Step 2, collect all filtratings (about 5L) heating and be concentrated into 1L.Attention heats while stirring, and prevents the sticking end;
Step 3, liquid concentrator are collected in the 1L beaker, and 70 ℃ of water-baths are concentrated into 400ml, under this temperature, and the solvent evaporates rapid speed, temperature is too high, then can influence active polysaccharide, prevents polysaccharide carbonization in the heat-processed.Need 24h;
Step 4, liquid concentrator branch are filled in the centrifuge tube of 8 50ml, and the centrifugal 10min of 4500 rpm obtains water-soluble Cordyceps polysaccharide solution;
Step 5, the about 400ml of collection supernatant remove insoluble impurities; To the 2L beaker; Slowly add (120ml/ minute) and go into 4 times of volumes (the Cordyceps polysaccharide biological activity that 4 times of volume ethanol obtain after handling is good) absolute ethyl alcohol; To ethanol final concentration (after the 400ml supernatant adds 1600ml ethanol; The TV of system is 2L, and 1600ml is exactly a final concentration divided by 2L) 75%, 4 ℃ leave standstill 12h.Notice that the limit adds the ethanol limit and stirs;
Step 6,2000ml precipitation solution branch are filled in 40 50ml centrifuge tubes, the centrifugal 10min of 4500rpm, and the supernatant that inclines, all alcohol precipitation polysaccharide are collected in 4 50ml centrifuge tubes;
Step 7,100% ethanol are washed throw out 2 times, the vacuum and low temperature lyophilize;
Step 8, worm summer in winter entomogenous fungi mycelium polysaccharide 5g is dissolved in the 500ml zero(ppm) water stirred overnight, centrifugal 5 minutes of 10000rpm; Obtain the 500ml supernatant, in supernatant, add 100% ethanol 500ml, the limit edged stirs; 4 ℃ of depositions are spent the night, centrifugal 5 minutes of 10000rpm, and deposition is the Cordyceps mycelium polysaccharide fraction; This moment, throw out obtained said Cordyceps mycelium polysaccharide powder through the vacuum and low temperature lyophilize, was placed in the dry environment and preserved.
Shown in accompanying drawing 1; Under the condition of 40 times of magnifications; (a) detects figure for the present invention at the killing tumor cell aspect of performance among Fig. 1; (b) is that prior art detects figure at the killing tumor cell aspect of performance among Fig. 1, and tumor cell line MCA205 (fibrosarcoma cell) is inoculated in cell cultures 12 orifice plates, and cell inoculation density is 1 * 10
5/ hole adds the Cordyceps mycelium polysaccharide simultaneously, and concentration is 0.2mg/ml, and polysaccharide is respectively this product (ccg-150 Cordyceps mycelium polysaccharide fraction) and currently available products (Cordyceps mycelium Crude polysaccharides).Cultivate after 3 days, microscopically is observed the growth of tumour cell situation, and the result shows, tumour cell mass mortality in this group of products, and cell debris is more, and tumour cell has part dead in of the present invention group, the growth of tumour cell in the lower left corner, the visual field is good.Through both comparative descriptions, the present invention has lethal effect preferably to tumour cell.
The foregoing description only is explanation technical conceive of the present invention and characteristics, and its purpose is to let the personage who is familiar with this technology can understand content of the present invention and enforcement according to this, can not limit protection scope of the present invention with this.All equivalences that spirit is done according to the present invention change or modify, and all should be encompassed within protection scope of the present invention.
Claims (4)
1. Cordyceps mycelium polysaccharide extracting process that is used for the prophylaxis of tumours compound formulation is characterized in that: may further comprise the steps:
Step 1, get the Chinese caterpillar fungus fermentation extracting solution and concentrate the bacterium powder 100g form, add 4L H
2O boils behind the 4h while hot filtered through gauze and removes residue;
Step 2, the filtrating heating that will obtain through step 1 be concentrated into 1L and obtain liquid concentrator;
Step 3, the liquid concentrator of step 2 is collected in the 1L beaker, 70 ℃ of water-baths are concentrated into 400ml;
Step 4, liquid concentrator branch in the step 3 is filled in the centrifuge tube of 8 50ml, under 4500 rpm centrifugation rates, handles 10min, obtain water-soluble Cordyceps polysaccharide solution;
Step 5, collect the about 400ml of supernatant in step 4 solution, the removal insoluble impurities to the 2L beaker, leaves standstill the acquisition precipitation solution after slowly adding 4 times of volume absolute ethyl alcohols;
Step 6, the precipitation solution branch of step 5 is filled in 40 50ml centrifuge tubes, alcohol precipitation polysaccharide in said each centrifuge tube is collected in 4 50ml centrifuge tubes after under 4500 rpm centrifugation rates, handling 10min;
Step 7, wash through the said alcohol precipitation polysaccharide of step 62 times through 100% ethanol, the vacuum and low temperature lyophilize obtains the Cordyceps mycelium polysaccharide;
Step 8, the Cordyceps mycelium polysaccharide of step 7 is dissolved in the zero(ppm) water; Stir after 2 hours; The 10000rpm centrifugation rate is handled down and was obtained supernatant in 5 minutes, in this supernatant, adds 100% ethanol, and the limit edged stirs; After 4 ℃ of depositions hour, under the 10000rpm centrifugation rate, handle again 5 minutes post precipitations at this moment throw out obtain said Cordyceps mycelium polysaccharide powder through the vacuum and low temperature lyophilize.
2. Cordyceps mycelium polysaccharide extracting process according to claim 1 is characterized in that: carry out stirring operation when filtrating is heated in the said step 2 simultaneously.
3. Cordyceps mycelium polysaccharide extracting process according to claim 1 is characterized in that: in the said step 2 in 70 ℃ of water-baths the hold-time be 24h.
4. Cordyceps mycelium polysaccharide extracting process according to claim 1 is characterized in that: dwell temperature is 4 ℃ in the said step 5, and the time is 12h.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103329733A (en) * | 2013-06-21 | 2013-10-02 | 夏西超 | Methods for preparing entomogenous fungus conjugate and extracting polysaccharides of entomogenous fungus conjugate |
| CN103435712A (en) * | 2013-08-27 | 2013-12-11 | 王辉 | Extraction method of cordyceps sinensis intracellular polysaccharide |
| CN108359027A (en) * | 2018-05-25 | 2018-08-03 | 浙江丹晖生物科技有限公司 | A kind of lentinan powder extraction process for treating tumour |
| CN110256591A (en) * | 2019-06-21 | 2019-09-20 | 东莞东阳光保健品研发有限公司 | A kind of Chinese caterpillar fungus polysaccharide extract and its preparation method and application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1482144A (en) * | 2003-07-25 | 2004-03-17 | 山东智灵生物工程有限公司 | Method for preparing galactomannanpeptide and product |
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2012
- 2012-04-18 CN CN201210113422.0A patent/CN102653567B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1482144A (en) * | 2003-07-25 | 2004-03-17 | 山东智灵生物工程有限公司 | Method for preparing galactomannanpeptide and product |
Non-Patent Citations (4)
| Title |
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| 《微生物学通报》 20060630 肖建辉等 "江西虫草发酵液脱色及其多糖提取分离的优化工艺" 24-30 1-4 第33卷, 第3期 * |
| 《科学大众（科学教育）》 20111020 仇冬梅等 "虫草菌丝体发酵液中多糖的提取和含量测定" 180 1-4 , 第10期 * |
| 仇冬梅等: ""虫草菌丝体发酵液中多糖的提取和含量测定"", 《科学大众（科学教育）》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103329733A (en) * | 2013-06-21 | 2013-10-02 | 夏西超 | Methods for preparing entomogenous fungus conjugate and extracting polysaccharides of entomogenous fungus conjugate |
| CN103435712A (en) * | 2013-08-27 | 2013-12-11 | 王辉 | Extraction method of cordyceps sinensis intracellular polysaccharide |
| CN108359027A (en) * | 2018-05-25 | 2018-08-03 | 浙江丹晖生物科技有限公司 | A kind of lentinan powder extraction process for treating tumour |
| CN110256591A (en) * | 2019-06-21 | 2019-09-20 | 东莞东阳光保健品研发有限公司 | A kind of Chinese caterpillar fungus polysaccharide extract and its preparation method and application |
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| CN102653567B (en) | 2014-06-18 |
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