CN104592336A - Method for extracting cordycepin in cordyceps militaris - Google Patents
Method for extracting cordycepin in cordyceps militaris Download PDFInfo
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- CN104592336A CN104592336A CN201410842829.6A CN201410842829A CN104592336A CN 104592336 A CN104592336 A CN 104592336A CN 201410842829 A CN201410842829 A CN 201410842829A CN 104592336 A CN104592336 A CN 104592336A
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Abstract
The invention provides a method for extracting cordycepin in cordyceps militaris. Cordycepin is a medicine with a natural source. The cordycepin has a good clinical application prospect besides of a plurality of pharmacologic actions of resisting tumors, resisting bacteria and viruses, carrying out immunoregulation and removing free radicals, but is always affected by the factors such as raw materials and purification process. The method comprises the following steps: fully crushing a cordyceps militaris culture medium or a bacteria sample, dissolving into water, carrying out ultrasonic treatment to improve the dissolving-out amount of the cordycepin; filtering to obtain a crude filtrate; and gathering and purifying the filtrate, and carrying out adsorption, elution and concentration on the filtrate through a reverse ODS filled column, so as to obtain the pure cordycepin crystal. The method has the beneficial effects that the extraction efficiency of the cordycepin can be improved; the extraction time is shortened; extraction and purification processes of the cordycepin are optimized; and the method is simple in processing steps and selected equipment, high in operability and low in energy consumption.
Description
Technical field
The invention belongs to medical art, especially relate to the extracting method of cordycepin in a kind of Cordyceps militaris (L.) Link..
Background technology
Cordyceps sinensis, has another name called cordyceps sinensis, also referred to as Cordyceps sinensis, is called for short Chinese caterpillar fungus.It is the rare traditional Chinese medicine of Chinese tradition, it is the Hepialus larva parasitized by the Cordyceps fungus of Hypocreales Clavicipitaceae Cordyceps in alpine meadow soil, larva is ossify, under optimum conditions, go out long bar-shaped stroma by bombys batryticatus head end pumping summer and form (i.e. the sporophore of Cordyceps fungus and bombys batryticatus sclerotium (larva corpse) form complex body).But worldwide Cordyceps sinensis distribution natural resource quantity is little, and in China, although Cordyceps sinensis kind is a lot, reach hundreds of, what be used as medicine just only has two kinds: Cordyceps fungus, Cordyceps militaris (L.) Link. bacterium.Cordycepin is a kind of medicine of natural origin, has outside the multiple pharmacological effect such as antitumor, anti-bacteria and anti-virus, immunomodulatory, scavenging free radicals, has good potential applicability in clinical practice.Due to the impact by the factor such as raw material, purge process, its price remains high always, and fancy price has become the bottleneck that restriction cordycepin develops.Therefore extraction process and its yield of raising of studying cordycepin have important economic worth and wide market application potential.
Summary of the invention
For solving the problems of the technologies described above, the invention provides the extracting method of cordycepin in a kind of Cordyceps militaris (L.) Link..
The technical solution used in the present invention is: the extracting method of cordycepin in a kind of Cordyceps militaris (L.) Link., comprises the following steps:
Step one, fully pulverizes Chinese caterpillar fungus culture medium or thalline sample, and add water mixing, supersound extraction under vacuum condition, obtains crude extract after being filtered by mixed solution.
Step 2, carries out settlement treatment by crude extract, collects supernatant liquor, carries out essence filter, obtain filtrate through millipore filtration.
Step 3, adsorbs filtrate through anti-phase ODS filled column, then uses water, methanol aqueous solution and methanol-eluted fractions successively, make pillar regenerate simultaneously, carry out HPLC detection to elutriant, collects the elutriant detecting and have cordycepin, obtains purification liquid.
Step 4, carries out rotary evaporation in vacuo to purification liquid, obtains evaporation concentration liquid, crystallization at evaporation concentration liquid is placed in 1 ~ 5 DEG C, is dissolved in purified water by the crystal obtained after suction filtration, carries out recrystallization, cordycepin crystal, finally by cordycepin crystal after frozen drying.
Preferably, in step one, the pH value of water is 4-5, and the volume ratio of Chinese caterpillar fungus culture medium or thalline sample and water is 1: 8-30.
Preferably, the ultrasonic number of times of step one is 2-5 time, each ultrasonic interval 10-15 minute.
Preferably, in step 2, the aperture of millipore filtration is 3-5 μm.
Preferably, step 3 filtrate is adsorbed through anti-phase ODS filled column, and often liter of filtrate adds the filler of 20-60g, then uses methanol aqueous solution and the methanol-eluted fractions of the water of 5-15 column volume, a 5-15 column volume successively, makes pillar regenerate simultaneously; DS packing material size is less than or equal to 150 μm.
The advantage that the present invention has and positively effect are: owing to adopting technique scheme, improve cordycepin extraction efficiency, reduce extraction time; Be optimized the Isolation and purification technique of cordycepin, processing step is simple with the equipment selected, and workable, energy consumption is low.
Embodiment
Below by specific embodiment, technical scheme of the present invention is further described.
Embodiment 1:
Step one, fully pulverizes Chinese caterpillar fungus culture medium or thalline sample, adds the water mixing that pH value is 4-5, the volume ratio of Chinese caterpillar fungus culture medium or thalline sample and water is 1: 8, supersound extraction under vacuum condition, supersound extraction 2 times, 10 minutes, interval, obtains crude extract after being filtered by mixed solution.
Step 2, carries out settlement treatment by crude extract, collects supernatant liquor, is that 3-5 μm of millipore filtration carries out essence filter, obtains filtrate through aperture.
Step 3, filtrate is adsorbed through anti-phase ODS filled column, DS packing material size is less than or equal to 150 μm, often liter of filtrate adds the filler of 20g, then use the water of 5 column volumes, the methanol aqueous solution of 5 column volumes and methanol-eluted fractions successively, make pillar regenerate simultaneously, HPLC detection is carried out to elutriant, collect the elutriant detecting and have cordycepin, obtain purification liquid.
Step 4, carries out rotary evaporation in vacuo to purification liquid, obtains evaporation concentration liquid, crystallization at evaporation concentration liquid is placed in 4 DEG C, is dissolved in purified water by the crystal obtained after suction filtration, carries out recrystallization, cordycepin crystal, finally by cordycepin crystal after frozen drying.
Embodiment 2:
Step one, fully pulverizes Chinese caterpillar fungus culture medium or thalline sample, adds the water mixing that pH value is 4-5, the volume ratio of Chinese caterpillar fungus culture medium or thalline sample and water is 1: 30, supersound extraction under vacuum condition, supersound extraction 5 times, 15 minutes, interval, obtains crude extract after being filtered by mixed solution.
Step 2, carries out settlement treatment by crude extract, collects supernatant liquor, is that 3-5 μm of millipore filtration carries out essence filter, obtains filtrate through aperture.
Step 3, filtrate is adsorbed through anti-phase ODS filled column, DS packing material size is less than or equal to 150 μm, often liter of filtrate adds the filler of 60g, then use the water of 15 column volumes, the methanol aqueous solution of 15 column volumes and methanol-eluted fractions successively, make pillar regenerate simultaneously, HPLC detection is carried out to elutriant, collect the elutriant detecting and have cordycepin, obtain purification liquid.
Step 4, carries out rotary evaporation in vacuo to purification liquid, obtains evaporation concentration liquid, crystallization at evaporation concentration liquid is placed in 4 DEG C, is dissolved in purified water by the crystal obtained after suction filtration, carries out recrystallization, cordycepin crystal, finally by cordycepin crystal after frozen drying.
Above to invention has been exemplary description; should be noted that; when not departing from core of the present invention, any simple distortion, amendment or other those skilled in the art can not spend the equivalent replacement of creative work all to fall into protection scope of the present invention.
Claims (5)
1. the extracting method of cordycepin in Cordyceps militaris (L.) Link., is characterized in that: comprise the following steps:
Step one, fully pulverizes Chinese caterpillar fungus culture medium or thalline sample, and add water mixing, supersound extraction under vacuum condition, obtains crude extract after being filtered by mixed solution;
Step 2, carries out settlement treatment by described crude extract, collects supernatant liquor, carries out essence filter, obtain filtrate through millipore filtration;
Step 3, adsorbs described filtrate through anti-phase ODS filled column, then uses water, methanol aqueous solution and methanol-eluted fractions successively, make pillar regenerate simultaneously, carry out HPLC detection to elutriant, collects the elutriant detecting and have cordycepin, obtains purification liquid;
Step 4, carries out rotary evaporation in vacuo to described purification liquid, obtains evaporation concentration liquid, crystallization at evaporation concentration liquid is placed in 1 ~ 5 DEG C, is dissolved in purified water by the crystal obtained after suction filtration, carries out recrystallization, cordycepin crystal, finally by described cordycepin crystal after frozen drying.
2. the extracting method of cordycepin in Cordyceps militaris (L.) Link. according to claim 1, it is characterized in that: in described step one, the pH value of water is 4-5, the volume ratio of described Chinese caterpillar fungus culture medium or described thalline sample and water is 1: 8-30.
3. the extracting method of cordycepin in Cordyceps militaris (L.) Link. according to claim 1, is characterized in that: the ultrasonic number of times of described step one is 2-5 time, each ultrasonic interval 10-15 minute.
4. the extracting method of cordycepin in Cordyceps militaris (L.) Link. according to claim 1, is characterized in that: in described step 2, the aperture of millipore filtration is 3-5 μm.
5. the extracting method of cordycepin in Cordyceps militaris (L.) Link. according to claim 1, it is characterized in that: described step 3 filtrate is adsorbed through anti-phase ODS filled column, often liter of described filtrate adds the filler of 20-60g, then use methanol aqueous solution and the methanol-eluted fractions of the water of 5-15 column volume, a 5-15 column volume successively, make pillar regenerate simultaneously, described methanol aqueous solution is the methanol aqueous solution of volume percent 5-30%, and DS packing material size is less than or equal to 150 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410842829.6A CN104592336A (en) | 2014-12-30 | 2014-12-30 | Method for extracting cordycepin in cordyceps militaris |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410842829.6A CN104592336A (en) | 2014-12-30 | 2014-12-30 | Method for extracting cordycepin in cordyceps militaris |
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| CN104592336A true CN104592336A (en) | 2015-05-06 |
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| CN201410842829.6A Pending CN104592336A (en) | 2014-12-30 | 2014-12-30 | Method for extracting cordycepin in cordyceps militaris |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105111264A (en) * | 2015-08-16 | 2015-12-02 | 深圳市倍昂生物科技有限公司 | 3'-deoxyadenosine crystal form and preparation method therefor |
| CN105942105A (en) * | 2016-06-01 | 2016-09-21 | 合肥丰瑞隆生物科技有限公司 | Cordyceps militaris beverage and making method thereof |
| CN108991193A (en) * | 2018-06-29 | 2018-12-14 | 浙江香仟科技有限公司 | A kind of Radix Glycyrrhizae lily Cordyceps militaris solid clearing lung-heat health preserving tea and preparation method |
| CN113952902A (en) * | 2021-11-19 | 2022-01-21 | 广东容大生物股份有限公司 | Method for extracting water-soluble cordycepin serving as antibacterial growth promoter |
-
2014
- 2014-12-30 CN CN201410842829.6A patent/CN104592336A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105111264A (en) * | 2015-08-16 | 2015-12-02 | 深圳市倍昂生物科技有限公司 | 3'-deoxyadenosine crystal form and preparation method therefor |
| CN105942105A (en) * | 2016-06-01 | 2016-09-21 | 合肥丰瑞隆生物科技有限公司 | Cordyceps militaris beverage and making method thereof |
| CN108991193A (en) * | 2018-06-29 | 2018-12-14 | 浙江香仟科技有限公司 | A kind of Radix Glycyrrhizae lily Cordyceps militaris solid clearing lung-heat health preserving tea and preparation method |
| CN113952902A (en) * | 2021-11-19 | 2022-01-21 | 广东容大生物股份有限公司 | Method for extracting water-soluble cordycepin serving as antibacterial growth promoter |
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Application publication date: 20150506 |