CN111171169A - Method for efficiently and simultaneously extracting polysaccharide and adenosine from fermented cordyceps sinensis powder - Google Patents
Method for efficiently and simultaneously extracting polysaccharide and adenosine from fermented cordyceps sinensis powder Download PDFInfo
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- CN111171169A CN111171169A CN202010025040.7A CN202010025040A CN111171169A CN 111171169 A CN111171169 A CN 111171169A CN 202010025040 A CN202010025040 A CN 202010025040A CN 111171169 A CN111171169 A CN 111171169A
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- adenosine
- cordyceps sinensis
- sinensis powder
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/167—Purine radicals with ribosyl as the saccharide radical
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Abstract
The invention discloses a method for efficiently and simultaneously extracting polysaccharide and adenosine from fermented cordyceps sinensis powder, which comprises the steps of extracting the fermented cordyceps sinensis powder at 95-100 ℃ for 2-4 hours to obtain an extracting solution, wherein the weight ratio of feed liquid is 1: 10-1: 15; centrifuging the extractive solution, and collecting supernatant; concentrating the supernatant, and measuring the relative density of the concentrated solution at 50-60 ℃ to be 0.98-1.18; adding absolute ethyl alcohol until the concentration of the ethanol is 40-85%, carrying out alcohol precipitation, centrifuging, separating to obtain a precipitate and a supernatant, and concentrating the supernatant into a thick paste; drying, and drying the precipitate and the soft extract respectively. The method is environment-friendly, the product is safe, the obtained product has high yield, safety and high adenosine content.
Description
Technical Field
The invention belongs to the technical field of biological extraction, and relates to a method for simultaneously extracting cordyceps polysaccharide and adenosine from fermented cordyceps sinensis powder.
Background
Cordyceps sinensis is a rare traditional Chinese medicine and has multiple effects. The fermented cordyceps sinensis powder is prepared by fermenting paecilomyces hepiali in cordyceps sinensis, has multiple components, wherein polysaccharide and adenosine are important effective components in the fermented cordyceps sinensis powder, and has wide physiological activity. At present, the cordyceps health-care products in the market take adenosine as a main functional component and polysaccharide as the second component. Cordyceps adenosine and polysaccharide are two effective components, and corious versicolor intracellular glycopeptide is a mixed component.
At present, most methods for extracting polysaccharide are water extraction and alcohol precipitation methods, and most adenosine are water extraction, alcohol extraction and enzymolysis methods. The alcohol extraction method has certain potential safety hazard and the utilization rate of the bacterial powder is low; the water extraction method has low adenosine content and contains a large amount of impurities. How to simply, efficiently and safely extract adenosine from cordyceps is an important research direction.
Disclosure of Invention
The invention aims to achieve the effect of simultaneously extracting the polysaccharide and the adenosine in the cordyceps by groping a simple water extraction and alcohol precipitation method, and has the advantages of simple and safe operation and high material utilization rate.
The purpose of the invention is realized by the following modes:
a method for efficiently and simultaneously extracting polysaccharide and adenosine from fermented cordyceps sinensis powder comprises the following steps:
extracting fermented cordyceps sinensis powder for 2-4 hours at the temperature of 95-100 ℃ to obtain an extracting solution, wherein the weight ratio of the material liquid to the material liquid is 1: 10-1: 15; centrifuging the extractive solution, and collecting supernatant; concentrating the supernatant, and measuring the relative density of the concentrated solution at 50-60 ℃ to be 0.98-1.18; adding absolute ethyl alcohol until the concentration of the ethanol is 40-85%, carrying out alcohol precipitation, centrifuging, separating to obtain a precipitate and a supernatant, and concentrating the supernatant into a thick paste; drying, and drying the precipitate and the soft extract respectively.
The drying temperature is preferably 60-70 ℃.
The preferable alcohol precipitation condition is 4 ℃ alcohol precipitation for 12-20 h.
The centrifugation is preferably carried out for 5min under the condition of 4000 r/min.
Preferably, the ethanol concentration adopted by the ethanol precipitation is 50-75%, and most preferably, the ethanol concentration adopted by the ethanol precipitation is 60%.
The relative density of the concentrated solution is 1.03-1.19 measured at 50-60 ℃. Preferably, the relative density of the concentrated solution is 1.022-1.181 measured at 50-60 ℃.
The inventor discovers that the total sugar in the cordyceps sinensis and other components have a coprecipitation phenomenon; when the density of the extracting solution is higher, the polysaccharide has adhesiveness, and other substances are caused to settle along with the polysaccharide, which is shown in that the precipitation yield is increased and the total sugar content is reduced.
The adenosine is a substance which can be dissolved in water and alcohol, and the polysaccharide is a substance which can be dissolved in water and can not be dissolved in alcohol. Two products with high yield are obtained by one extraction method, the utilization rate of the bacterial powder is increased, and the yield is improved.
Compared with the prior art, the invention has the beneficial effects that: in the method, the content of polysaccharide in the precipitate is more than 30 percent, and the content of adenosine in the supernatant is more than 12 mg/g. The product is prepared into solid and is suitable for industrial production; polysaccharide and adenosine can be simultaneously extracted from the fermented cordyceps sinensis powder, and the utilization rate of the fermented cordyceps sinensis powder is high; the water extraction and alcohol precipitation method has simple and safe process.
The detection method comprises the following steps: the method for detecting cordyceps polysaccharide as water-soluble polysaccharide and the method for detecting cordyceps adenosine as national standard adenosine.
Detailed Description
The present invention is further illustrated by the following specific examples, wherein the fermented Cordyceps powder used is purchased from Jiangsu Shenhua pharmaceutical Co.
Example 1: water extraction. Taking 1kg of fermented cordyceps sinensis powder, adding 10 times of water, and extracting for 2.5h at 95-100 ℃. Centrifuging the extractive solution at 4000r/min for 5min, collecting supernatant, concentrating into soft extract, and oven drying at 70 deg.C to obtain sample.
Example 2: alcohol extraction method. Taking 1kg of fermented cordyceps sinensis powder, adding 10 times of 25%, 50% and 75% ethanol, and heating and refluxing for extraction for 2.5 h. Centrifuging the extractive solution at 4000r/min for 5min, collecting supernatant, concentrating into soft extract, and oven drying at 70 deg.C to obtain sample.
Example 3: water extraction and alcohol precipitation. Taking 1kg of fermented cordyceps sinensis powder, adding 10 times of water, and extracting for 2.5h at 95-100 ℃. Centrifuging the extractive solution at 4000r/min for 5min, collecting supernatant, concentrating to obtain concentrated solution with relative density of 1.102 (measured at 50-60 deg.C), adding anhydrous ethanol to make final ethanol concentration be 60%, precipitating with ethanol at 4 deg.C for 12h, centrifuging the precipitated solution at 4000r/min for 5min, collecting supernatant, concentrating into soft extract, and oven drying the soft extract and precipitate in 70 deg.C oven respectively.
TABLE 1 Effect of extraction method on effective ingredients of Cordyceps
| Extraction method | The yield is% | Polysaccharide% | Adenosine mg/g |
| Mushroom powder | 4.5 | 3.4 | |
| Water extraction method | 34.1 | 17.7 | 7.9 |
| Alcohol extraction method 25% | 29.9 | 16.50 | 8.55 |
| Alcohol extraction method 50% | 27.8 | 11.26 | 9.81 |
| Alcohol extraction method 75% | 23.3 | 4.9 | 11.4 |
| Water extraction and alcohol precipitation (precipitation) | 16.6 | 33.5 | 0.4 |
| Extracting with water and precipitating with ethanol (supernatant) | 17.1 | 3.4 | 14.1 |
As can be seen from the data, the higher the ethanol concentration in the alcohol extraction method, the lower the polysaccharide content, and the higher the adenosine content. The total yield of the sediment and the supernatant in the water extraction and alcohol precipitation method is similar to that of the water extraction method, and the content of polysaccharide in the sediment and the content of adenosine in the supernatant are higher than that of the water extraction method and the alcohol extraction method. The water extraction and alcohol precipitation method is obviously superior to the water extraction method and the alcohol extraction method.
Example 4: taking 10kg of fermented cordyceps sinensis powder, adding 10 times of water, and extracting for 2.5h at 95-100 ℃. Extracting at 4000r/min, centrifuging for 5min, collecting supernatant, dividing the supernatant into 8 parts, concentrating to different densities, adding 100% ethanol to make the final ethanol concentration be 60%, precipitating with ethanol at 4 deg.C for 12h, subjecting the precipitated solution to 4000r/min, centrifuging for 5min, collecting supernatant, concentrating to obtain soft extract, and oven drying the soft extract and precipitate in a 70 deg.C oven to obtain the sample. When the density is higher, the adenosine content is higher, and the density is higher than 1.2, the supernatant yield and the adenosine content tend to be stable values.
TABLE 2 Effect of concentration on the active ingredients of fermented Cordyceps
| Density of | The precipitation yield is% | Precipitated polysaccharide% | The yield of the supernatant is% | Supernatant adenosine mg/g |
| 0.977 | 10.0 | 55.1 | 23.2 | 11.1 |
| 0.996 | 12.1 | 47.1 | 20.3 | 11.8 |
| 1.022 | 14.4 | 40.7 | 16.9 | 12.2 |
| 1.077 | 16.8 | 36.0 | 16.1 | 13.9 |
| 1.132 | 20.5 | 31.1 | 12.6 | 15.8 |
| 1.181 | 22.2 | 27.7 | 8.9 | 18.5 |
| 1.227 | 24.5 | 24.4 | 6.6 | 21.2 |
| 1.241 | 25.1 | 23.6 | 6.1 | 21.9 |
Example 5: taking 10kg of cordyceps sinensis powder, adding 10 times of water, and extracting for 2.5h at 95-100 ℃. Extracting at 4000r/min, centrifuging for 5min, collecting supernatant, concentrating the concentrated solution to density of 1.071, dividing the concentrated solution into 10 parts, adding 100% ethanol to make the final ethanol concentration be 40%, 50%, 60%, 70%, 80%, precipitating with ethanol at 4 deg.C for 12h, precipitating with ethanol at 4000r/min, centrifuging for 5min, collecting supernatant, concentrating into slurry, precipitating, and oven drying at 70 deg.C to obtain the sample. When alcohol precipitation is carried out, the concentration of ethanol is increased, the content of adenosine is higher, the concentration of ethanol is higher than 80%, and the yield of supernate and the content of adenosine tend to be stable values.
TABLE 3 influence of ethanol concentration on the active ingredients of fermented Cordyceps
| Concentration of ethanol | The precipitation yield is% | Precipitated polysaccharide% | The yield of the supernatant is% | Supernatant adenosine mg/g |
| 40％ | 12.1 | 31.9 | 20.7 | 10.2 |
| 45％ | 13.4 | 32.8 | 20.3 | 11.1 |
| 50％ | 13.8 | 34.3 | 20.0 | 11.5 |
| 55％ | 14.9 | 35.6 | 19.3 | 12.1 |
| 60％ | 15.3 | 36.8 | 18.1 | 12.7 |
| 65％ | 16.5 | 35.1 | 16.5 | 13.6 |
| 70％ | 17.3 | 33.9 | 15.9 | 14.6 |
| 75％ | 18.4 | 31.3 | 15.1 | 15.2 |
| 80％ | 20.1 | 28.8 | 14.2 | 16.1 |
| 85％ | 20.7 | 26.9 | 13.8 | 16.5 |
The above examples are merely illustrative for clarity and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description.
Claims (8)
1. A method for efficiently and simultaneously extracting polysaccharide and adenosine from fermented cordyceps sinensis powder is characterized by comprising the following steps:
extracting fermented cordyceps sinensis powder for 2-4 hours at the temperature of 95-100 ℃ to obtain an extracting solution, wherein the weight ratio of the material liquid to the material liquid is 1: 10-1: 15; centrifuging the extractive solution, and collecting supernatant; concentrating the supernatant, and measuring the relative density of the concentrated solution at 50-60 ℃ to be 0.98-1.18; adding absolute ethyl alcohol until the concentration of the ethanol is 40-85%, carrying out alcohol precipitation, centrifuging, separating to obtain a precipitate and a supernatant, and concentrating the supernatant into a thick paste; drying, and drying the precipitate and the soft extract respectively.
2. The method for efficiently and simultaneously extracting the polysaccharide and the adenosine from the fermented cordyceps sinensis powder according to claim 1, wherein the drying temperature is 60-70 ℃.
3. The method for efficiently and simultaneously extracting the polysaccharides and the adenosine from the fermented cordyceps sinensis powder according to claim 1, wherein the alcohol precipitation condition is 4 ℃ alcohol precipitation for 12-20 hours.
4. The method for simultaneously extracting the polysaccharide and the adenosine from the fermented cordyceps sinensis powder with high efficiency according to claim 1, wherein the centrifugation is carried out for 5min under the condition of 4000 r/min.
5. The method for efficiently and simultaneously extracting the polysaccharide and the adenosine from the fermented cordyceps sinensis powder according to claim 1, wherein the ethanol concentration adopted by the ethanol precipitation is 50-75%.
6. The method for efficiently and simultaneously extracting the polysaccharides and the adenosine from the fermented cordyceps sinensis powder according to claim 5, wherein the ethanol concentration adopted by the ethanol precipitation is 60%.
7. The method for efficiently and simultaneously extracting the polysaccharide and the adenosine from the fermented cordyceps sinensis powder according to claim 1, wherein the relative density of the concentrated solution is 1.03-1.19 measured at 50-60 ℃.
8. The method for efficiently and simultaneously extracting the polysaccharide and the adenosine from the fermented cordyceps sinensis powder according to claim 7, wherein the relative density of the concentrated solution is 1.022-1.181 when measured at 50-60 ℃.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101229199A (en) * | 2008-01-15 | 2008-07-30 | 重庆和润生物工程有限公司 | Integrative extract method of multi-active ingredient in cordyceps militaris mycelium |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101229199A (en) * | 2008-01-15 | 2008-07-30 | 重庆和润生物工程有限公司 | Integrative extract method of multi-active ingredient in cordyceps militaris mycelium |
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Application publication date: 20200519 |