CN106467566A - The enzymolysis technique of cordycepin and the application in health food in the Cordyceps militaris of Daxing'an Mountainrange - Google Patents
The enzymolysis technique of cordycepin and the application in health food in the Cordyceps militaris of Daxing'an Mountainrange Download PDFInfo
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- cordycepin
- cordyceps militaris
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- KQLDDLUWUFBQHP-UHFFFAOYSA-N Cordycepin Natural products C1=NC=2C(N)=NC=NC=2N1C1OCC(CO)C1O KQLDDLUWUFBQHP-UHFFFAOYSA-N 0.000 title claims abstract description 55
- OFEZSBMBBKLLBJ-BAJZRUMYSA-N cordycepin Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)C[C@H]1O OFEZSBMBBKLLBJ-BAJZRUMYSA-N 0.000 title claims abstract description 55
- OFEZSBMBBKLLBJ-UHFFFAOYSA-N cordycepine Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)CC1O OFEZSBMBBKLLBJ-UHFFFAOYSA-N 0.000 title claims abstract description 55
- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 50
- 238000000034 method Methods 0.000 title claims abstract description 34
- 235000013402 health food Nutrition 0.000 title claims abstract description 8
- 238000009210 therapy by ultrasound Methods 0.000 claims abstract description 17
- 239000012141 concentrate Substances 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 7
- 238000004108 freeze drying Methods 0.000 claims abstract description 7
- 239000008280 blood Substances 0.000 claims abstract description 4
- 210000004369 blood Anatomy 0.000 claims abstract description 4
- 230000000845 anti-microbial effect Effects 0.000 claims abstract description 3
- 230000001442 anti-mosquito Effects 0.000 claims abstract description 3
- 230000001203 anti-plasmodial effect Effects 0.000 claims abstract description 3
- 239000004599 antimicrobial Substances 0.000 claims abstract description 3
- 208000015181 infectious disease Diseases 0.000 claims abstract description 3
- 244000052769 pathogen Species 0.000 claims abstract description 3
- 230000001717 pathogenic effect Effects 0.000 claims abstract description 3
- 230000001737 promoting effect Effects 0.000 claims abstract description 3
- 230000001629 suppression Effects 0.000 claims abstract description 3
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract 4
- 235000012000 cholesterol Nutrition 0.000 claims abstract 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000012153 distilled water Substances 0.000 claims description 16
- 239000000843 powder Substances 0.000 claims description 16
- 239000000287 crude extract Substances 0.000 claims description 14
- 239000000284 extract Substances 0.000 claims description 12
- 239000000706 filtrate Substances 0.000 claims description 12
- 108090000145 Bacillolysin Proteins 0.000 claims description 11
- 108010059892 Cellulase Proteins 0.000 claims description 11
- 102000035092 Neutral proteases Human genes 0.000 claims description 11
- 108091005507 Neutral proteases Proteins 0.000 claims description 11
- 229940106157 cellulase Drugs 0.000 claims description 11
- 238000010298 pulverizing process Methods 0.000 claims description 11
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 238000005119 centrifugation Methods 0.000 claims description 9
- 241000190633 Cordyceps Species 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 239000000919 ceramic Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 5
- 238000007710 freezing Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 5
- 230000010355 oscillation Effects 0.000 claims description 5
- 238000001556 precipitation Methods 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 238000012360 testing method Methods 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 3
- 206010028980 Neoplasm Diseases 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 9
- 238000004458 analytical method Methods 0.000 abstract description 5
- 238000004811 liquid chromatography Methods 0.000 abstract description 5
- 230000000259 anti-tumor effect Effects 0.000 abstract description 3
- 241000196324 Embryophyta Species 0.000 description 5
- 239000003814 drug Substances 0.000 description 4
- 241000233866 Fungi Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 206010005963 Bone formation increased Diseases 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 208000005171 Dysmenorrhea Diseases 0.000 description 1
- 206010013935 Dysmenorrhoea Diseases 0.000 description 1
- 208000019695 Migraine disease Diseases 0.000 description 1
- CKZWLFLXOQDVEX-UHFFFAOYSA-N [2-(6-aminopurin-9-yl)-5-[[[2-(6-aminopurin-9-yl)-5-(hydroxymethyl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]oxolan-3-yl] [5-(6-aminopurin-9-yl)-4-hydroxyoxolan-2-yl]methyl hydrogen phosphate Chemical compound C1=NC2=C(N)N=CN=C2N1C1OC(COP(O)(=O)OC2C(OC(CO)C2)N2C3=NC=NC(N)=C3N=C2)CC1OP(O)(=O)OCC(CC1O)OC1N1C(N=CN=C2N)=C2N=C1 CKZWLFLXOQDVEX-UHFFFAOYSA-N 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 230000002443 hepatoprotective effect Effects 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 206010027599 migraine Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses the enzymolysis technique of cordycepin and the application in health food in a kind of Cordyceps militaris of Daxing'an Mountainrange, it passes sequentially through selection, pulverize, dissolving, enzymolysis, ultrasonic Treatment, high voltage pulse is processed, preliminary filtration, concentrate, vacuum lyophilization, the present invention is extracted using the method for enzymolysis, can be good at improving extraction ratio, and combine ultrasonic Treatment, high-voltage pulse processes and carries out aid in treatment, extraction ratio can be greatly improved, record cordycepin content checking in extracting solution through liquid chromatography analysis, the extraction ratio of the cordycepin of the present invention can reach 0.852%, this health promoting product is applied to antitumor, anti-microbial pathogen, suppression protozoon, reduce cholesterol, blood sugar lowering, infection, in antiplasmodial and anti-mosquito.
Description
Technical field
The present invention relates to the enzymolysis technique of cordycepin and the application in health food in a kind of Cordyceps militaris of Daxing'an Mountainrange,
Belong to health care technology field.
Background technology
Daxinganling forest area Wild Medicinal animal and plant resource enriches, and is that development forest zone substitutes industry, cultivates new economic increasing
The importance of long point.Wherein, the Cordyceps militaris containing in Daxinganling forest area, can reduce cylinder super-oxidation lipid and free radical water
Flat, delay organ and whole body aging.Cordyceps militaris are a kind of edible fungi liked by people very much in recent years, with famous Chinese medicine
Cordyceps have similar pharmacologically active.
Cordycepin also known as Cordycepin, Cordycepin, cordycepin, be in Cordyceps and Cordyceps militaris (L.) Link. (especially
Ucleosides) main active is also first nucleoside antibiotics separated from funguses.And Daxing'an Mountainrange Cordyceps militaris
In same contain abundant cordycepin, a kind of medicine of natural origin of cordycepin, there are Cordyceps in traditional Chinese medical science medical knowledge the same
Negative and positive are with the function of mending with two-ways regulation total balance of the body;It hepatoprotective, protect kidney, in terms of lung moistening due to the purer effect of composition more preferably,
And strongly invigorating QI and blood, the disease such as incurable dysmenorrhea, migraine, hyperosteogeny now can be eliminated.In terms of doctor trained in Western medicine medical knowledge angle
Cordycepin has antitumor, defying age, antibacterial, antiviral, immunomodulating, improves metabolism, removes multiple medicines such as free radical
Reason acts on, and has good potential applicability in clinical practice.The research of cordycepin is now just becoming pharmaceutical chemistry, defying age, beauty treatment, health care at present
An extremely active field in product field.
Therefore, how efficiently to extract cordycepin from the Cordyceps militaris of Daxing'an Mountainrange, for grass element research with answer apparatus
There is extremely important effect.
Content of the invention
The present invention be directed to existing technical problem, provide in a kind of Cordyceps militaris of Daxing'an Mountainrange digest cordycepin technique and
In the application in health food it is therefore an objective to efficiently extract cordycepin from Cordyceps militaris, intend to solve asking of prior art presence
Topic.
For achieving the above object, the present invention provides following technical scheme：Cordycepin is digested in a kind of Cordyceps militaris of Daxing'an Mountainrange
Technique it is characterised in that it comprises the following steps：
（1）Choose Daxing'an Mountainrange Cordyceps militaris, and dried, then dried Daxing'an Mountainrange Cordyceps militaris are carried out at pulverizing
Reason, and cross 20 mesh sieves；
（2）Enzymolysis：Cordyceps militaris powder after pulverizing is added in distilled water, adds neutral protease while stirring, be placed in
In 35-45 DEG C of water-bath, digest 40min, add cellulase afterwards, be placed in 40-50 DEG C of water-bath, then digest
30min；
（3）Ultrasonic Treatment：By described step（2）The enzymolysis solution obtaining carries out supersound process 20min；
（4）High voltage pulse is processed：Solution after ultrasonic Treatment is placed in pulse in high voltage pulse and processes 10min, pulse is
Big pressure is 350Mpa, and minimum pressure is 20Mpa, and pulse frequency is 5-10Hz；
（5）Preliminary filtration：By described step（4）The solution obtaining is heated to 100 degrees Celsius, and is incubated 15min, using ceramic membrane
Filtered, repeat to filter 2-4 time, merging filtrate；
（6）Concentrate：By described step（5）The filtrate obtaining is placed in Rotary Evaporators and carries out concentration；
（7）Vacuum lyophilization：By described step（6）Concentrated solution after concentration is placed in vacuum freezing instrument that to carry out vacuum cold
Freeze, obtain cordycepin crude extract.
Further, preferably, described step（7）It is additionally provided with essence afterwards and put forward step, specially：Cordycepin will be obtained thick
Extract is dissolved in distilled water, adds activated carbon in thick cordycepin crude extract solution, and constant temperature oscillation 2.5h at 30 DEG C
, it is subsequently placed in centrifugation 15min in the centrifuge of 5000r/min, abandons precipitation, add Sevage liquid afterwards, extract 35min, then
It is placed in centrifugation 15min in the centrifuge of 4200r/min, takes the superiors' clear liquid, repeat aforesaid operations 3-5 time, then be added thereto to
The dehydrated alcohol of the 95% of 5 times of volumes, puts into 4 DEG C of refrigerator standing 24h, takes out and be centrifuged in the centrifuge of 4800r/min
20min, abandoning supernatant, drying precipitated as cordycepin extract.
Further, preferably, described step（1）During dried, it is to be carried out drying to containing using 60 DEG C of hot run test
Water rate is less than 4%, and is pulverized using plant pulverizer.
Further, preferably, described step（2）In, Cordyceps militaris powder：Neutral protease：The ratio of cellulase is
100:2:3.5.
Further, preferably, described step（2）In, the ratio of Cordyceps militaris powder and distilled water is 100:1ml/g.
Further, preferably, during described ultrasonic Treatment, ultrasonic power 500W, frequency 26 kHz.
Additionally, the invention provides a kind of health food made using cordycepin, it adopts in the Cordyceps militaris of Daxing'an Mountainrange
The technique of enzymolysis cordycepin is prepared from, this enzymolysis：Cordyceps militaris powder after pulverizing is added in distilled water, adds while stirring
Neutral protease, is placed in 35-45 DEG C of water-bath, digests 40min, adds cellulase afterwards, be placed in 40-50 DEG C
In water-bath, then digest 30min；
（3）Ultrasonic Treatment：By described step（2）The enzymolysis solution obtaining carries out supersound process 20min；
（4）High voltage pulse is processed：Solution after ultrasonic Treatment is placed in pulse in high voltage pulse and processes 10min, pulse is
Big pressure is 350Mpa, and minimum pressure is 20Mpa, and pulse frequency is 5-10Hz；
（5）Preliminary filtration：By described step（4）The solution obtaining is heated to 100 degrees Celsius, and is incubated 15min, using ceramic membrane
Filtered, repeat to filter 2-4 time, merging filtrate；
（6）Concentrate：By described step（5）The filtrate obtaining is placed in Rotary Evaporators and carries out concentration；
（7）Vacuum lyophilization：.
Compared with prior art, the invention has the beneficial effects as follows：
The present invention is extracted using the method for enzymolysis, can be good at improving extraction ratio, and combines ultrasonic Treatment, high-tension pulse
Punching processes and carries out aid in treatment, can greatly improve extraction ratio, record cordycepin in extracting solution through liquid chromatography analysis
Content is verified, the extraction ratio of the cordycepin of the present invention can reach 0.852%.
Specific embodiment
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described,
Obviously, described embodiment is only a part of embodiment of the present invention, rather than whole embodiments.Based in the present invention
Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under the premise of not making creative work, all
Belong to the scope of protection of the invention.
Embodiment one
The technique of cordycepin is digested it is characterised in that it comprises the following steps in a kind of Cordyceps militaris of Daxing'an Mountainrange：
（1）Choose Daxing'an Mountainrange Cordyceps militaris, and dried, then dried Daxing'an Mountainrange Cordyceps militaris are carried out at pulverizing
Reason, and cross 20 mesh sieves, wherein, during dried, it is to be dried using 60 DEG C of hot run test to be less than 4% to moisture content, and utilizes
Plant pulverizer is pulverized.
（2）Enzymolysis：Cordyceps militaris powder after pulverizing is added in distilled water, adds neutral protease, juxtaposition while stirring
In 45 DEG C of water-bath, digest 40min, add cellulase afterwards, be placed in 40 DEG C of water-bath, then digest 30min, its
In, Cordyceps militaris powder：Neutral protease：The mass ratio of cellulase is 100:2:3.5, the ratio of Cordyceps militaris powder and distilled water
Example is 100:1ml/g；
（3）Ultrasonic Treatment：By described step（2）The enzymolysis solution obtaining carries out supersound process 20min, ultrasonic power 500W,
Frequency 26 kHz；
（4）High voltage pulse is processed：Solution after ultrasonic Treatment is placed in pulse in high voltage pulse and processes 10min, pulse is
Big pressure is 350Mpa, and minimum pressure is 20Mpa, and pulse frequency is 8Hz；
（5）Preliminary filtration：By described step（4）The solution obtaining is heated to 100 degrees Celsius, and is incubated 15min, using ceramic membrane
Filtered, repeated filter 23, merging filtrate；
（6）Concentrate：By described step（5）The filtrate obtaining is placed in Rotary Evaporators and carries out concentration；
（7）Vacuum lyophilization：By described step（6）Concentrated solution after concentration is placed in vacuum freezing instrument that to carry out vacuum cold
Freeze, obtain cordycepin crude extract.
（8）Essence puies forward step, specially：Cordycepin crude extract will be obtained be dissolved in distilled water, to thick cordycepin crude extract
Activated carbon is added in solution, and constant temperature oscillation 2.5h at 30 DEG C, it is subsequently placed in centrifugation in the centrifuge of 5000r/min
15min, abandons precipitation, adds Sevage liquid afterwards, extracts 35min, then is placed in centrifugation in the centrifuge of 4200r/min
15min, takes the superiors' clear liquid, repeats aforesaid operations 5 times, then be added thereto to 5 times of volumes 95% dehydrated alcohol, put into 4 DEG C
Refrigerator standing 24h, take out in the centrifuge of 4800r/min be centrifuged 20min, abandoning supernatant, drying precipitated be Cordycepses
Plain extract.
Record cordycepin content checking, the extraction ratio of the cordycepin of the present embodiment in extracting solution through liquid chromatography analysis
Can reach 0.852%.
Embodiment two
The technique of cordycepin is digested it is characterised in that it comprises the following steps in a kind of Cordyceps militaris of Daxing'an Mountainrange：
（1）Choose Daxing'an Mountainrange Cordyceps militaris, and dried, then dried Daxing'an Mountainrange Cordyceps militaris are carried out at pulverizing
Reason, and cross 20 mesh sieves, wherein, during dried, it is to be dried using 60 DEG C of hot run test to be less than 4% to moisture content, and utilizes
Plant pulverizer is pulverized.
（2）Enzymolysis：Cordyceps militaris powder after pulverizing is added in distilled water, adds neutral protease, juxtaposition while stirring
In 40 DEG C of water-bath, digest 40min, add cellulase afterwards, be placed in 40 DEG C of water-bath, then digest 30min, its
In, Cordyceps militaris powder：Neutral protease：The mass ratio of cellulase is 100:2:3.5, the ratio of Cordyceps militaris powder and distilled water
Example is 100:1ml/g；
（3）Ultrasonic Treatment：By described step（2）The enzymolysis solution obtaining carries out supersound process 20min, ultrasonic power 500W,
Frequency 26 kHz；
（4）High voltage pulse is processed：Solution after ultrasonic Treatment is placed in pulse in high voltage pulse and processes 10min, pulse is
Big pressure is 350Mpa, and minimum pressure is 20Mpa, and pulse frequency is 5Hz；
（5）Preliminary filtration：By described step（4）The solution obtaining is heated to 100 degrees Celsius, and is incubated 15min, using ceramic membrane
Filtered, repeat to filter 4 times, merging filtrate；
（6）Concentrate：By described step（5）The filtrate obtaining is placed in Rotary Evaporators and carries out concentration；
（7）Vacuum lyophilization：By described step（6）Concentrated solution after concentration is placed in vacuum freezing instrument that to carry out vacuum cold
Freeze, obtain cordycepin crude extract.
（8）Essence puies forward step, specially：Cordycepin crude extract will be obtained be dissolved in distilled water, to thick cordycepin crude extract
Activated carbon is added in solution, and constant temperature oscillation 2.5h at 30 DEG C, it is subsequently placed in centrifugation in the centrifuge of 5000r/min
15min, abandons precipitation, adds Sevage liquid afterwards, extracts 35min, then is placed in centrifugation in the centrifuge of 4200r/min
15min, takes the superiors' clear liquid, repeats aforesaid operations 5 times, then be added thereto to 5 times of volumes 95% dehydrated alcohol, put into 4 DEG C
Refrigerator standing 24h, take out in the centrifuge of 4800r/min be centrifuged 20min, abandoning supernatant, drying precipitated be Cordycepses
Plain extract.
Record cordycepin content checking, the extraction ratio of the cordycepin of the present embodiment in extracting solution through liquid chromatography analysis
Can reach 0.855%.
Embodiment three
The technique of cordycepin is digested it is characterised in that it comprises the following steps in a kind of Cordyceps militaris of Daxing'an Mountainrange：
（1）Choose Daxing'an Mountainrange Cordyceps militaris, and dried, then dried Daxing'an Mountainrange Cordyceps militaris are carried out at pulverizing
Reason, and cross 20 mesh sieves, wherein, during dried, it is to be dried using 60 DEG C of hot run test to be less than 4% to moisture content, and utilizes
Plant pulverizer is pulverized.
（2）Enzymolysis：Cordyceps militaris powder after pulverizing is added in distilled water, adds neutral protease, juxtaposition while stirring
In 35 DEG C of water-bath, digest 40min, add cellulase afterwards, be placed in 42 DEG C of water-bath, then digest 30min, its
In, Cordyceps militaris powder：Neutral protease：The mass ratio of cellulase is 100:2:3.5, the ratio of Cordyceps militaris powder and distilled water
Example is 100:1ml/g；
（3）Ultrasonic Treatment：By described step（2）The enzymolysis solution obtaining carries out supersound process 20min, ultrasonic power 500W,
Frequency 26 kHz；
（4）High voltage pulse is processed：Solution after ultrasonic Treatment is placed in pulse in high voltage pulse and processes 10min, pulse is
Big pressure is 350Mpa, and minimum pressure is 20Mpa, and pulse frequency is 8Hz；
（5）Preliminary filtration：By described step（4）The solution obtaining is heated to 100 degrees Celsius, and is incubated 15min, using ceramic membrane
Filtered, repeated filter 23, merging filtrate；
（6）Concentrate：By described step（5）The filtrate obtaining is placed in Rotary Evaporators and carries out concentration；
（7）Vacuum lyophilization：By described step（6）Concentrated solution after concentration is placed in vacuum freezing instrument that to carry out vacuum cold
Freeze, obtain cordycepin crude extract.
（8）Essence puies forward step, specially：Cordycepin crude extract will be obtained be dissolved in distilled water, to thick cordycepin crude extract
Activated carbon is added in solution, and constant temperature oscillation 2.5h at 30 DEG C, it is subsequently placed in centrifugation in the centrifuge of 5000r/min
15min, abandons precipitation, adds Sevage liquid afterwards, extracts 35min, then is placed in centrifugation in the centrifuge of 4200r/min
15min, takes the superiors' clear liquid, repeats aforesaid operations 4 times, then be added thereto to 5 times of volumes 95% dehydrated alcohol, put into 4 DEG C
Refrigerator standing 24h, take out in the centrifuge of 4800r/min be centrifuged 20min, abandoning supernatant, drying precipitated be Cordycepses
Plain extract.
Record cordycepin content checking, the extraction ratio of the cordycepin of the present embodiment in extracting solution through liquid chromatography analysis
Can reach 0.858%.
Additionally, the invention provides a kind of health food made using cordycepin, it adopts in the Cordyceps militaris of Daxing'an Mountainrange
The technique of enzymolysis cordycepin is prepared from, and this health promoting product is applied to antitumor, anti-microbial pathogen, suppression protozoon, reduces gallbladder admittedly
In alcohol, blood sugar lowering, infection, antiplasmodial and anti-mosquito.
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, permissible
Understand and can carry out multiple changes, modification, replacement to these embodiments without departing from the principles and spirit of the present invention
And modification, the scope of the present invention be defined by the appended.
Claims (7)
1. digest the technique of cordycepin in a kind of Cordyceps militaris of Daxing'an Mountainrange it is characterised in that it comprises the following steps：
（1）Choose Daxing'an Mountainrange Cordyceps militaris, and dried, then dried Daxing'an Mountainrange Cordyceps militaris are carried out at pulverizing
Reason, and cross 20 mesh sieves；
（2）Enzymolysis：Cordyceps militaris powder after pulverizing is added in distilled water, adds neutral protease while stirring, be placed in
In 35-45 DEG C of water-bath, digest 40min, add cellulase afterwards, be placed in 40-50 DEG C of water-bath, then digest
30min；
（3）Ultrasonic Treatment：By described step（2）The enzymolysis solution obtaining carries out supersound process 20min；
（4）High voltage pulse is processed：Solution after ultrasonic Treatment is placed in pulse in high voltage pulse and processes 10min, pulse is
Big pressure is 350Mpa, and minimum pressure is 20Mpa, and pulse frequency is 5-10Hz；
（5）Preliminary filtration：By described step（4）The solution obtaining is heated to 100 degrees Celsius, and is incubated 15min, using ceramic membrane
Filtered, repeat to filter 2-4 time, merging filtrate；
（6）Concentrate：By described step（5）The filtrate obtaining is placed in Rotary Evaporators and carries out concentration；
（7）Vacuum lyophilization：By described step（6）Concentrated solution after concentration is placed in vacuum freezing instrument that to carry out vacuum cold
Freeze, obtain cordycepin crude extract.
2. in a kind of Cordyceps militaris of Daxing'an Mountainrange according to claim 1 digest cordycepin technique it is characterised in that：Described
Step（7）It is additionally provided with essence afterwards and put forward step, specially：Cordycepin crude extract will be obtained be dissolved in distilled water, to thick Cordycepses
Activated carbon is added in plain crude extract solution, and constant temperature oscillation 2.5h at 30 DEG C, it is subsequently placed in the centrifuge of 5000r/min
Middle centrifugation 15min, abandons precipitation, afterwards add Sevage liquid, extract 35min, then be placed in the centrifuge of 4200r/min from
Heart 15min, takes the superiors' clear liquid, repeats aforesaid operations 3-5 time, then be added thereto to 5 times of volumes 95% dehydrated alcohol, put
Enter 4 DEG C refrigerator standing 24h, take out in the centrifuge of 4800r/min be centrifuged 20min, abandoning supernatant, drying precipitated be
Cordycepin extract.
3. in a kind of Cordyceps militaris of Daxing'an Mountainrange according to claim 1 digest cordycepin technique it is characterised in that：Described
Step（1）During dried, it is to be dried using 60 DEG C of hot run test to be less than 4% to moisture content, and is entered using plant pulverizer
Row is pulverized.
4. in a kind of Cordyceps militaris of Daxing'an Mountainrange according to claim 1 digest cordycepin technique it is characterised in that：Described
Step（2）In, Cordyceps militaris powder：Neutral protease：The mass ratio of cellulase is 100:2:3.5.
5. in a kind of Cordyceps militaris of Daxing'an Mountainrange according to claim 1 digest cordycepin technique it is characterised in that：Described
Step（2）In, the ratio of Cordyceps militaris powder and distilled water is 100:1ml/g.
6. the technique digesting cordycepin in a kind of Daxing'an Mountainrange Cordyceps militaris according to claim 1-5 any one, it is special
Levy and be：During described ultrasonic Treatment, ultrasonic power 500W, frequency 26 kHz.
7. a kind of health food made using cordycepin it is characterised in that：Described cordycepin is any using claim 1-6
The technique digesting cordycepin in a kind of Daxing'an Mountainrange Cordyceps militaris described in one is prepared from, and this health promoting product is applied to anti-swollen
In tumor, anti-microbial pathogen, suppression protozoon, reduction cholesterol, blood sugar lowering, infection, antiplasmodial and anti-mosquito.
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| CN105294395A (en) * | 2015-09-23 | 2016-02-03 | 安徽阳光药业有限公司 | Method for preparing cordycepic acid and cordycepin by simultaneous extraction-combination with column chromatography-crystallization purification |
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