CN104478978A - Extraction method of adenosine in scarlet caterpillar fungus - Google Patents
Extraction method of adenosine in scarlet caterpillar fungus Download PDFInfo
- Publication number
- CN104478978A CN104478978A CN201410732004.9A CN201410732004A CN104478978A CN 104478978 A CN104478978 A CN 104478978A CN 201410732004 A CN201410732004 A CN 201410732004A CN 104478978 A CN104478978 A CN 104478978A
- Authority
- CN
- China
- Prior art keywords
- adenosine
- extraction
- precipitation
- link
- supernatant liquor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
- C07H1/08—Separation; Purification from natural products
Abstract
The invention provides an extraction method of adenosine in scarlet caterpillar fungus. The extraction method specifically comprises the following steps: (1) superfine grinding; (2) ultrasonic bionic extraction and enzymatic extraction of adenosine in scarlet caterpillar fungus. The extraction method has the beneficial effects that the hard cell wall of scarlet caterpillar fungus is broken by applying the enzymolysis technology, thus being beneficial to dissolution of cellular contents, being beneficial to smooth operation of the extraction process, increasing the extraction rate of adenosine while ensuring the bioactivity of adenosine, and reducing the waste; adenosine extracted by applying the method not only has high extraction rate and is easy to separate and purify subsequently but also is easy to absorb and can give full play to the effects of adenosine in vivo.
Description
Technical field
the present invention relates to technical field of plant extraction, particularly relate to a kind of extracting method of Cordyceps militaris (L.) Link. adenosine.
Background technology
cordyceps militaris (L.) Link. has another name called Cordyceps militaris (L.) Link., Cordyceps militaris etc., belongs to xenogenesis together with Cordyceps sinensis, is the type species of Cordyceps, belongs to Ascomycotina Cordyceps, is that it colonizes in the complex body of stroma and the ossified pupal cell that Insecta lepidopteran noctuid pupal cell grows.Years of researches show that Cordyceps militaris (L.) Link. has immunomodulatory, the multiple pharmacologically active such as antitumor, antiviral, anti-infective, are widely used in tumour, immunologic hypofunction, comprise in the various diseases such as HIV (human immunodeficiency virus) infection.
adenosine is the acid of a kind of endogenous purine nucleoside, and have physiologically active widely, in Cordyceps militaris (L.) Link., the content of adenosine is similar to Cordyceps sinensis, even higher, how more than 0.01%.So far from Chinese caterpillar fungus and fermented hypha, isolation identification has 11 nucleoside compounds, comprises VITAMIN B4, adenosine, uridylic, uridine, guanine, guanosine, xanthoglobulin, inosine, thymus pyrimidine, Thymine deoxyriboside and uridylic deoxynucleoside etc.Cordycepin is main active ingredient in Cordyceps militaris (L.) Link., is also first nucleoside antibiotics separated from fungi, also known as cordycepin, cordycepin.The molecular formula of cordycepin is C10H13N5O3, and relative molecular mass is 251.Cordyceps militaris (L.) Link. plants the bacterial classification that uniquely can form cordycepin in Cordyceps in a large number reported at present more than 300.In natural cordyceps or fermentation mycelium, cordycepin does not exist or extremely trace.Be studied in detail for its structure and pharmacological action both at home and abroad at present, research proves that cordycepin has multiple biological activity, is mainly manifested in several aspects such as antibacterial, anti-inflammatory, antitumor and immunomodulatory.Wherein cordycepin is the focus that vast scientists is paid close attention to the restraining effect of tumour always.
Summary of the invention
the object of this invention is to provide a kind of extracting method of Cordyceps militaris (L.) Link. adenosine.
object of the present invention is achieved through the following technical solutions: a kind of extracting method of Cordyceps militaris (L.) Link. adenosine, and the concrete steps of its extracting method are:
(1) micronizing: take the Cordyccps-militaris-(L.)-link. Sporophore that 200g is dried, put into supper micron mill and pulverize 5-15 minute;
(2) ultrasonic bionic extraction, the enzymolysis and extraction of Cordyceps militaris (L.) Link. adenosine, comprising:
a, according to solid-liquid ratio, every g material adds the extraction agent of 1ml, be that the ratio of 6 ~ 25m/v adds the hydrochloric acid soln that pH value is 4.0-6.0, take the wall breaking enzyme that mass volume ratio is 0.5 ~ 1%, under 37 DEG C of conditions, be incubated 10 ~ 30min, then in ultrasonic wave with after power heating l ~ 5 clock of 60% ~ 80%, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying;
b, again according to solid-liquid ratio, every g material adds the extraction agent of 1ml, be 6 ~ 25m/v ratio to above-mentioned pH value be 4.0-6.0 hydrochloric acid soln extract, adding pH in the precipitation of air-drying after finishing is 6 ~ 7.5, and concentration is the buffer solution of sodium phosphate of 0.1mol/L, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying collects supernatant liquor, precipitation natural air drying;
6 ~ 30 times of volumes are added in the precipitation of c, the most backward natural air drying, every g material adds the extraction agent of 1ml, pH value is 8.0 ~ 9.0 alkaline solution of sodium hydroxide, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, discard precipitation;
d, merge three supernatant liquors, get 100ul after crossing the water system film of 0.45um and add ultrapure water and be diluted to 1000ul, after crossing the water system film of 0.22um, in high performance liquid chromatograph, measure the content of adenosine, obtain Cordyceps militaris (L.) Link. adenosine.
the present invention has following beneficial effect
1. the method simulation human gastrointestinal tract acid or alkali environment carries out the extraction of Cordyceps militaris (L.) Link. adenosine, and the adenosine obtained is easy to be processed into various types of medicament; After the formulation that process for processing becomes oral liquid or other types, the direct absorption of Cordyceps militaris (L.) Link. effective constituent can be guaranteed, alleviate stomach burden;
2. the method extraction Cordyceps militaris (L.) Link. adenosine content is high, the content of adenosine can reach every g crude drug and extract the amount obtaining 2.825mg, the extraction yield of final activeconstituents can up to 0.304%, all higher than other general extraction methods extraction yield, and quality controllable, material damages few, with short production cycle, be applicable to scale operation, market potential is huge;
3. zymolysis technique has abolished the hard cell walls of Cordyceps militaris (L.) Link., is conducive to the stripping of entocyte, is conducive to carrying out smoothly of leaching process, ensureing adenosine bioactive while, improves the rate of promoting of adenosine, cuts the waste;
4. apply the method and extract the adenosine obtained, not only extraction yield is high, is easy to follow-up separation and purification, and is easy to absorb, and can give full play to adenosine effect in vivo.
Embodiment
embodiment 1
an extracting method for Cordyceps militaris (L.) Link. adenosine, the concrete steps of its extracting method are:
(1) micronizing: take the Cordyccps-militaris-(L.)-link. Sporophore that 200g is dried, puts into supper micron mill and pulverizes 10 minutes.
(2) the ultrasonic bionic extraction of Cordyceps militaris (L.) Link. adenosine adds enzymolysis and extraction, comprising:
a, being 15 (m/v) according to solid-liquid ratio (every g material adds the extraction agent of 1ml), ratio adds the hydrochloric acid soln that pH value is 5.0, take the wall breaking enzyme that mass volume ratio is 0.5%, 20min is incubated under 37 DEG C of conditions, then in ultrasonic wave with after power heating l ~ 5 clock of 60% ~ 80%, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying;
b, again according to solid-liquid ratio (every g material adds the extraction agent of 1ml) be 15 (m/v) ratio to above-mentioned pH value be 5.0 hydrochloric acid soln extract.Adding pH in the precipitation of air-drying after finishing is 7.0, concentration is the buffer solution of sodium phosphate of 0.1mol/L, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying collects supernatant liquor, precipitation natural air drying;
the pH value adding 15 times of volumes (every g material adds the extraction agent of 1ml) in the precipitation of c, the most backward natural air drying is 8.5 alkaline solution of sodium hydroxide, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, discard precipitation;
d, merge three supernatant liquors, get 100ul after crossing the water system film of 0.45um and add ultrapure water and be diluted to 1000ul, in high performance liquid chromatograph, after crossing the water system film of 0.22um, measure the content of adenosine.
the content of adenosine can reach every g crude drug and extract the amount obtaining 2.265mg, and the extraction yield of final activeconstituents can up to 0.272%.
embodiment 2
an extracting method for Cordyceps militaris (L.) Link. adenosine, the concrete steps of its extracting method are:
(1) micronizing: take the Cordyccps-militaris-(L.)-link. Sporophore that 200g is dried, puts into supper micron mill and pulverizes 15 minutes.
(2) the ultrasonic bionic extraction of Cordyceps militaris (L.) Link. adenosine adds enzymolysis and extraction, comprising:
a, being 10 (m/v) according to solid-liquid ratio (every g material adds the extraction agent of 1ml), ratio adds the hydrochloric acid soln that pH value is 5.0, take the wall breaking enzyme that mass volume ratio is 1%, 20min is incubated under 37 DEG C of conditions, then in ultrasonic wave with after power heating l ~ 5 clock of 60% ~ 80%, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying;
b, again according to solid-liquid ratio (every g material adds the extraction agent of 1ml) be 10 (m/v) ratio to above-mentioned pH value be 4.0-6.0 hydrochloric acid soln extract.Adding pH in the precipitation of air-drying after finishing is 7.0, concentration is the buffer solution of sodium phosphate of 0.1mol/L, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying collects supernatant liquor, precipitation natural air drying;
the pH value adding 10 times of volumes (every g material adds the extraction agent of 1ml) in the precipitation of c, the most backward natural air drying is 8.5 alkaline solution of sodium hydroxide, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, discard precipitation;
d, merge three supernatant liquors, get 100ul after crossing the water system film of 0.45um and add ultrapure water and be diluted to 1000ul, in high performance liquid chromatograph, after crossing the water system film of 0.22um, measure the content of adenosine.
the content of adenosine can reach every g crude drug and extract the amount obtaining 2.825mg, and the extraction yield of final activeconstituents can up to 0.304%.
Claims (1)
1. an extracting method for Cordyceps militaris (L.) Link. adenosine, is characterized in that: the concrete steps of its extracting method are:
(1) micronizing: take the Cordyccps-militaris-(L.)-link. Sporophore that 200g is dried, put into supper micron mill and pulverize 5-15 minute;
(2) ultrasonic bionic extraction, the enzymolysis and extraction of Cordyceps militaris (L.) Link. adenosine, comprising:
A, according to solid-liquid ratio, every g material adds the extraction agent of 1ml, be that the ratio of 6 ~ 25m/v adds the hydrochloric acid soln that pH value is 4.0-6.0, take the wall breaking enzyme that mass volume ratio is 0.5 ~ 1%, under 37 DEG C of conditions, be incubated 10 ~ 30min, then in ultrasonic wave with after power heating l ~ 5 clock of 60% ~ 80%, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying;
B, again according to solid-liquid ratio, every g material adds the extraction agent of 1ml, be 6 ~ 25m/v ratio to above-mentioned pH value be 4.0-6.0 hydrochloric acid soln extract, adding pH in the precipitation of air-drying after finishing is 6 ~ 7.5, and concentration is the buffer solution of sodium phosphate of 0.1mol/L, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, precipitation natural air drying collects supernatant liquor, precipitation natural air drying;
6 ~ 30 times of volumes are added in the precipitation of c, the most backward natural air drying, every g material adds the extraction agent of 1ml, pH value is 8.0 ~ 9.0 alkaline solution of sodium hydroxide, with after power heating l ~ 5 clock of 60% ~ 80% in ultrasonic wave, with the centrifugal force of 8000g ~ 12000g under room temperature centrifugal 10 ~ 30 minutes, collect supernatant liquor, discard precipitation;
D, merge three supernatant liquors, get 100ul after crossing the water system film of 0.45um and add ultrapure water and be diluted to 1000ul, after crossing the water system film of 0.22um, in high performance liquid chromatograph, measure the content of adenosine, obtain Cordyceps militaris (L.) Link. adenosine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410732004.9A CN104478978A (en) | 2014-12-06 | 2014-12-06 | Extraction method of adenosine in scarlet caterpillar fungus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410732004.9A CN104478978A (en) | 2014-12-06 | 2014-12-06 | Extraction method of adenosine in scarlet caterpillar fungus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104478978A true CN104478978A (en) | 2015-04-01 |
Family
ID=52753599
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410732004.9A Pending CN104478978A (en) | 2014-12-06 | 2014-12-06 | Extraction method of adenosine in scarlet caterpillar fungus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104478978A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105362302A (en) * | 2015-11-25 | 2016-03-02 | 苏州普罗达生物科技有限公司 | Ultra-micro wall-breaking pulverizing method for cordyceps militaris |
| CN108948115A (en) * | 2018-10-16 | 2018-12-07 | 安徽祥康生物工程有限公司 | A kind of method of high efficiency extraction Cordyceps militaris adenosine |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102558265A (en) * | 2012-01-04 | 2012-07-11 | 正源堂（天津）生物科技有限公司 | Extract method of cordyceps militaris adenosine |
-
2014
- 2014-12-06 CN CN201410732004.9A patent/CN104478978A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102558265A (en) * | 2012-01-04 | 2012-07-11 | 正源堂（天津）生物科技有限公司 | Extract method of cordyceps militaris adenosine |
Non-Patent Citations (4)
| Title |
|---|
| 江晓路等，: "蛹虫草活性成分的测定.", 《食用菌学报》 * |
| 王志博等，: "用酶法对啤酒酵母细胞破壁优化条件的研究.", 《东北农业大学学报》 * |
| 赫功元等，: "银杏花粉不同破壁方法的比较分析.", 《南京林业大学学报（自然科学版》 * |
| 赵鲁杭等，: "《分子医学实验技术》", 30 April 2014 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105362302A (en) * | 2015-11-25 | 2016-03-02 | 苏州普罗达生物科技有限公司 | Ultra-micro wall-breaking pulverizing method for cordyceps militaris |
| CN108948115A (en) * | 2018-10-16 | 2018-12-07 | 安徽祥康生物工程有限公司 | A kind of method of high efficiency extraction Cordyceps militaris adenosine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tuli et al. | Pharmacological and therapeutic potential of Cordyceps with special reference to Cordycepin | |
| CN103432193A (en) | Microwave-assisted aqueous two-phase extraction and separation method of kudzu root total flavones | |
| CN102838686B (en) | Method for preparing high-purity agaricus blazei murrill polysaccharide | |
| CN102349924B (en) | Centella asiatica triterpenic acid single-glucopyranoside composition, its preparation method, its quantitative analysis method and its application | |
| CN103467617A (en) | Method for continuous counter-current ultrasonic extraction of high-purity astragalus polysaccharide | |
| CN106146684A (en) | The isolation and purification method of ganoderma spove powder GL-B | |
| CN101492706A (en) | Method for improving cordyceps sinensis bacterium native volume of production with cordyceps militaris link liquid fermentation | |
| CN105504079A (en) | Process for producing astragalus polysaccharide by using ultrasonic technology | |
| CN108997443A (en) | A kind of method that microbial fermentation extracts saponin(e in plant | |
| CN102936609A (en) | Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein | |
| CN103146592A (en) | Microzyme converting ginsenoside Rb1 to generate Rd and application thereof | |
| CN104357332B (en) | Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid | |
| CN105400845A (en) | Technology for producing radix astragali polysaccharide by utilization of microbial fermentation technology | |
| CN106008647B (en) | A kind of extracting method of Momordica charantial | |
| CN104497158A (en) | Method for extracting jujube polysaccharide | |
| CN104478978A (en) | Extraction method of adenosine in scarlet caterpillar fungus | |
| CN103342668A (en) | Simple method for extracting natural taurine from abalone viscera | |
| CN104783089A (en) | Preparation method of jujube extract | |
| CN104151385A (en) | Method for extracting cyclic adenosine monophosphate and application of cyclic adenosine monophosphate | |
| CN106619782A (en) | Herba rhodiolae extract and healthy product containing herba rhodiolae extract | |
| Zhang et al. | Enhancement of diosgenin production in Dioscorea zingiberensis cell culture by oligosaccharide elicitor from its endophytic fungus Fusarium oxysporum Dzf17 | |
| CN101367883B (en) | Method for extracting glossy ganoderma polysaccharide from fermentation liquor | |
| Tuli et al. | Cordycepin: A cordyceps metabolite with promising therapeutic potential | |
| CN103627770A (en) | Method used for biotransformation of pseudo-ginseng using two paecilomyces funguses | |
| CN110551167A (en) | Method for preparing astragaloside |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150401 |
|
| WD01 | Invention patent application deemed withdrawn after publication |