CN109825540A - A kind of preparation method and applications of the non-carbohydrate extract of Inonotus obliquus - Google Patents
A kind of preparation method and applications of the non-carbohydrate extract of Inonotus obliquus Download PDFInfo
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Abstract
The invention discloses a kind of preparation method and applications of the non-carbohydrate extract of Inonotus obliquus, the preparation method includes the culture and fermentation of Inonotus obliquus, the cold-induction of culture solution is handled, the preparation of freeze-dried powder, after primary extraction liquid is obtained by extraction to freeze-dried powder progress chloroform, dialysis treatment is carried out, high performance liquid chromatography is finally carried out and is purified, obtain extract.Above-mentioned preparation method is simple, period is short, the main component for the non-carbohydrate extract being prepared is s-triazine derivative, there is excellent bacteriostasis efficacy to bacterium and fungi, belong to biological agent, environmental pollution is few, it can be applied to fungicide, the antibacterial medicines of preparation environmental protection, it has a good application prospect, and sufficiently develops the application value of Inonotus obliquus, improve the utilization rate of this medicinal fungi.
Description
Technical field
The present invention relates to the preparations of a kind of medicinal fungi extract more particularly to a kind of non-carbohydrate extract of Inonotus obliquus
Method and its application.
Background technique
Currently, Inonotus obliquus Inonotus obliquus, is also known as inonotus obliquus, it is a kind of medicinal fungi of high value,
Belong to Basidiomycotina (Basidiomycotina), Hymenomycetes (Hymenomycetes), Aphyllophorales
(Aphyllophorales), Hymenochaetales (Hymenoehaetaeeae).Happiness is cold, is distributed across the domestomycetes in frigid zone, is born in white
Under the bark of the live standing trees such as birch, silvery birch, elm, alder or after felling trees it is dried-up on, and can cause white birch, silver-colored poplar, elm,
The white rot of alder.Its area for being mainly distributed on 45 °~50 ° of Northern Hemisphere north latitude, as northern North america, Finland, Poland, Russia,
The ground such as Chinese Heilungkiang, Jilin Province Changbaishan area, Hokkaido, Japan.
At abroad, coming across Russia and Poland using early stage to Inonotus obliquus, it be used to treat a variety of difficult and complicated cases,
Such as cancer, diabetes have numerous reports about the extraction and application study of Inonotus obliquus chemical component at present, main to concentrate
In antitumor, hypoglycemic, reducing blood lipid the research of the polysaccharose substance of Inonotus obliquus, due to the intracorporal active constituent of Inonotus obliquus
Type is more, and application value is high, and this field is studied and underuses and develop the non-saccharide active component of Inonotus obliquus body at present
Value, also lack the extraction preparation method of corresponding non-saccharide active component.
Therefore, the prior art need further to develop.
Summary of the invention
In view of the above technical problems, the embodiment of the invention provides a kind of preparation sides of the non-carbohydrate extract of Inonotus obliquus
Method and its application, the method is easy to operate, and the non-carbohydrate extract of the Inonotus obliquus prepared has good bacteriostatic activity,
It has a good application prospect.
The present invention provides following technical schemes:
The present invention provides a kind of preparation methods of the non-carbohydrate extract of Inonotus obliquus comprising following steps:
S1, Inonotus obliquus thallus is inoculated in fluid nutrient medium, after 22-27 DEG C of shake culture 1-2d, obtains seed
Seed liquor is inoculated in fluid nutrient medium, shake culture 5-7d, shaking speed 130- by liquid according to the inoculum concentration of 5-10%
160rpm/min obtains culture solution；
S2, culture solution is subjected to centrifugal treating, isolated thallus and fermentation liquid is deposited in respectively under 0-8 DEG C of environment and carried out
Cold-induction 20-40d；This step is induction Inonotus obliquus under adverse environment, expresses special active material.
S3, by after thallus and fermentation liquid that cold-induction is handled mix, carry out vacuum freeze drying, mixing will be lyophilized
Object is pulverized, and freeze-dried powder is obtained；
S4, freeze-dried powder is subjected to chloroform extraction, obtains primary extraction liquid, filter paper filtering is being carried out to primary extraction liquid, is being gone
Except impurity；
S5, dialysis treatment is carried out to filtered primary extraction liquid, after going the macromolecular of 500Da or more, obtains secondary and mention
Take liquid；This step passes through the polysaccharide or other macromolecular chaff interferences removed in primary extraction liquid in large quantity of dialysing.
S6, HPLC chromatogram purifying, isolated extract are carried out.The non-carbohydrate extract of the extract, that is, Inonotus obliquus.
In the preparation method, in step S1, the fluid nutrient medium of use includes each component of following mass percent:
Birch powder 5%, 10% glucose, 1.5% peptone, 0.2%KH2PO4, 0.2%MgSO4, remaining is water.
In the preparation method, in step S3, the condition of vacuum freeze drying are as follows: by the mixture of thallus and fermentation liquid
After being placed in -20 DEG C of freezing 2-3h, then after freezing 4-8h under the conditions of being placed in -80 DEG C, mixture taking-up is placed in vacuum freeze drying
It is dry in machine, until obtaining freeze-drying mixture.
In the preparation method, chloroform extraction methods are as follows: soxhlet's extraction method is used, using chloroform as solvent to freeze-dried powder
It is dissolved and is extracted, impregnate 10-12h, flowed back after 6-8h, obtain primary extraction liquid.
In the preparation method, in S6 step, the chromatographic column used is Hypersil GOLD C18 column, column temperature 28-
32℃。
In the preparation method, in S6 step, using acetonitrile-water eluent system, the volume of water and acetonitrile in eluant, eluent
Than are as follows: 1:(6-10), the flow velocity of secondary extracting solution is 0.2ml/min.
The present invention also provides a kind of non-carbohydrate extracts using Inonotus obliquus prepared by above-mentioned preparation method.
It is found through Mass Spectrometer Method, the main component of the non-carbohydrate extract is s-triazine derivative, this is for the first time in birch
Discovery s-triazine compound is extracted in brown pore fungi, and finds that such extract has excellent fungistatic effect, is had wide
Application prospect.
The present invention also provides the non-carbohydrate extract of above-mentioned Inonotus obliquus prepare fungicide, answering in antibacterial medicines
With.
The invention has the following advantages:
The present invention provides a kind of preparation method of the non-carbohydrate extract in new Inonotus obliquus, this method is for non-
Designed by the property of carbohydrate extract, different from the tedious steps of existing Inonotus obliquus extract, operation is simple, adopts
Extractant type is few, and extraction step time-consuming is short, and extraction effect is good；Pass through the master for the non-carbohydrate extract that this method obtains
Wanting ingredient is s-triazine derivative, this is that s-triazine derivative is extracted in this field from microbial body (Inonotus obliquus) for the first time
Object, s-triazine compound mainly uses chemical synthesis mode to obtain before this.Also, it was found that the non-carbohydrate extract is with excellent
Good bacteria resistance function has bacteriostasis efficacy, institute to pathogenic bacteria Escherichia coli, staphylococcus glucose coccus, candida albicans bacterium etc.
Stating non-carbohydrate extract can be applied to prepare fungicide, antibacterial medicines, not only have a good application prospect, and sufficiently open
The application value for having sent out Inonotus obliquus improves the utilization rate of this medicinal fungi.
Detailed description of the invention
Fig. 1 is that the high-efficient liquid phase chromatogram of Inonotus obliquus chloroform extract is composed；
Fig. 2 is the mass-spectrogram of Inonotus obliquus chloroform extract.
Specific embodiment
Below in conjunction with attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that is retouched
The embodiment stated is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, originally
Field technical staff every other embodiment obtained without creative efforts, belongs to protection of the present invention
Range.
Unless otherwise defined, technical and scientific term all used in this specification is led with technology of the invention is belonged to
The normally understood meaning of the technical staff in domain is identical.Used term is only in the description of the invention in this specification
The purpose of description specific embodiment is not intended to the limitation present invention.Term "and/or" used in this specification includes
Any and all combinations of one or more related listed items.In addition, invention described below difference is implemented
Technical characteristic involved in mode can be combined with each other as long as they do not conflict with each other.
The preparation of the non-carbohydrate extract of 1 Inonotus obliquus of embodiment
1, the culture and fermentation of Inonotus obliquus
Culture medium prescription: birch powder 5%, 10% glucose, 1.5% peptone, 0.2%KH2PO4, 0.2%MgSO4,
Yu Weishui.The fluid nutrient medium of configuration is subjected to high pressure sterilization processing.
The mycelium of Inonotus obliquus is inoculated in 50ml fluid nutrient medium, is shaken and is cultivated with 150rpm/min at 25 DEG C
Obtain seed liquor after 1-2d, seed liquor be inoculated in the culture medium of 300ml according to the inoculum concentration of 5-10%, at 25 DEG C with
150rpm/min vibration culture 5-7d, obtains culture solution.
2, the cold-induction of Inonotus obliquus
Above-mentioned Inonotus obliquus culture solution is placed in progress cold-induction 20-40d in 4 DEG C of refrigerators.Preferably, the cold-induction time is
30 days.This step facilitates Inonotus obliquus inducing expression target product in extreme circumstances.
3, the preparation of freeze-dried powder
By the fermentation liquid (including mycelium pellet and fermentation liquid) of cold-induction processing, vacuum freeze drying is carried out, it is mixed that will be lyophilized
It closes object to pulverize, obtains primary treatment product.For abundant broken wall, 150U/ can be added into the mixed liquor of thallus and fermentation liquid
The cytase of ml is tentatively digested, and carries out vacuum freeze drying again after handling 20min.It can also direct thallus in other embodiments
Freeze-drying process is carried out with the mixed liquor of fermentation liquid.
The method of vacuum freeze drying are as follows: after the mixed liquor of thallus and fermentation liquid is placed in -20 DEG C of freezing 2h, then be placed in -
After freezing 8h under the conditions of 80 DEG C, taking-up is placed in drying in vacuum freeze drier, until obtaining freeze-drying mixture；Freeze-drying is mixed
Object is pulverized, and freeze-dried powder is obtained.
4, the extraction of active constituent
Using soxhlet's extraction method, freeze-dried powder is dissolved and extracted using chloroform as solvent, impregnates 12h, after the 6h that flows back,
Primary extraction liquid is obtained, filter paper is reused and is filtered, removes impurity.
5, to the dialysis treatment of primary extraction liquid
One section of bag filter is taken, molecular cut off 500Da pricks one end cotton cord extremely, after other end addition 5ml filtering
Primary extraction liquid, and sealed, be suspended from bag filter in the beaker for filling distilled water, promoted with magnetic stirrer molten
Liquid exchange, until dialysis equilibrium.After above-mentioned dialysis treatment, the polysaccharide molecule in primary extraction liquid is removed,
Obtain secondary extracting solution.
6, purification process
High-efficient liquid phase chromatogram purification processing is carried out to secondary extracting solution, uses Hypersil GOLD C18 column (2.1
×100mm,5μm；Thermo Fisher Scientific, Waltham, USA), column temperature be 30 DEG C, secondary extracting solution with
The flow velocity of 0.2ml/min is carried out in the nanometer liquid phase systems of bidimensional.In elution process, eluant, eluent A uses water, and eluant, eluent B is second
Nitrile eluent system, in the volume ratio of two kinds of eluant, eluents are as follows: under the elution requirement of 1:8, chromatographic retention 1.3min collects purifying
Product, i.e. Inonotus obliquus chloroform extract, chromatogram are shown in Fig. 1.
7, mass spectral analysis
Chloroform extract is analyzed by mass spectrometry, Electrospray Ionization Mass Spectrometry carries out within the scope of the nucleocytoplasmic ratio of 50-1500m/z
Cation and anion full scan, nitrogen is as atomization gas, flow velocity 6L/min；Temperature is 180 DEG C；Pressure is 1.0Bar,
Data acquisition and processing (DAP) uses the LC/MS Data Analysis Software (edition 4 .0) provided with instrument.It is formed corresponding to specific element
Nucleocytoplasmic ratio data calculated using the formula predictions software provided with instrument.It is required that the standard of the nucleocytoplasmic ratio and substance measured
Error between nucleocytoplasmic ratio is no more than 5ppm.Mass spectral results are shown in Fig. 2, by the analysis to mass-spectrogram, learn that maximum peak is point
Daughter ion peak, M/Z value are 203, and the molecular formula of main active is C9H7N4O2, molecular weight 203, binding analysis other
Fragment ion peak finally obtains: compound C9H7N4O2, include 1,3,5- s-triazine rings, and be connected on the ring 4- nitro and
One phenyl ring, therefore can deduce that the compound is s-triazine derivative.This is isolated one kind from Inonotus obliquus for the first time
S-triazine derivative.
While above-mentioned experiment, separately set following control group: extraction material is identical with step, only saves cold-induction processing step
Suddenly, the Inonotus obliquus chloroform extract extracted to the control group detects, and finds: not finding compound in extract
C9H7N4O2, this explanation: handled without cold-induction, Inonotus obliquus does not express compound C9H7N4O2。
The bacteriostatic experiment of the non-carbohydrate extract of 2 Inonotus obliquus of embodiment
1, the culture of indicator bacteria:
Escherichia coli, staphylococcus glucose coccus, salmonella typhimurium, shigella dysenteriae are inoculated in beef extract egg respectively
After white peptone base, the shake culture under the conditions of 37 DEG C；After candida albicans bacterium is inoculated in PDA culture medium, shaken under the conditions of 28 DEG C
Culture is swung, bacterium solution is respectively obtained.
2, the setting of sample sets:
Experimental group and control group are respectively set in accordance with the following methods, every group of setting at least three repeats:
Experimental group 1: Inonotus obliquus chloroform extract is prepared according to above-mentioned full experiment step, as sample；Experimental group
2: sample will be used as after the fermented supernatant fluid low temperature concentration of filtering removal Inonotus obliquus thallus through everfermentation, cold-induction processing；It is real
Test group 3: it is essentially identical with 1 method of experimental group, cold-induction processing step is only saved, collects extract as sample.Positive control
Group: using streptomysin as sample.The sample of each experimental group is separately added into sterile distilled water dissolution, is configured to the molten of 10mg/ml
Liquid is filtered through 0.22 μm of sterilised membrane filter.
3, Antibacterial Activity
200uL bacterium suspension is added in solid medium tablets and is coated with uniformly, is made into plate containing bacterium, two experimental groups
In, sterilized filter paper is put into the sterile solution of extract impregnates 30min respectively, presss from both sides out filter paper patch with sterilizing tweezers
In on respectively plate containing bacterium；In positive controls, plate containing bacterium is handled using the filter paper for impregnating streptomysin 30min, each group repeats
Three times, it is placed in constant incubator to be inverted to cultivate and takes out plate afterwards for 24 hours, to the plate of experimental group and control group, be respectively adopted ten
Word interior extrapolation method measures antibacterial circle diameter size, takes number average value, test result is as follows table 1.
The calculation method of bacteriostasis rate:
Bacteriostasis rate=[(reagent agent antibacterial circle diameter-filter paper diameter)/(positive medicament antibacterial circle diameter-filter paper
Diameter)] × 100%
1 bacteriostasis rate test result of table
From above-mentioned antibacterial test result it is found that the Inonotus obliquus chloroform extract of experimental group 1 is to bacterium (Escherichia coli, gold
Yellow glucose coccus, shigella dysenteriae) and fungi (candida albicans bacterium) all have excellent bacteriostatic activity；And without supercooling
The experimental group 3 of induction, antibacterial test result can not show a candle to experimental group 1, this illustrates that chloroform extract bacteriostatic active ingredients are cold
It is expressed under inductive condition, cold-induction is important experimental procedure；Comparative experiments group 2 and experimental group 1 illustrate chloroform essence of the invention
The extraction effect of the extracting method of extract is good, and antipathogenic composition recovery rate is high, bacteriostatic activity is high.
It, can according to the technique and scheme of the present invention and this hair it is understood that for those of ordinary skills
Bright design is subject to equivalent substitution or change, and all these changes or replacement all should belong to the guarantor of appended claims of the invention
Protect range.
Claims (8)
1. a kind of preparation method of the non-carbohydrate extract of Inonotus obliquus, which comprises the following steps:
S1, Inonotus obliquus thallus is inoculated in fluid nutrient medium, after 22-27 DEG C of shake culture 1-2d, obtains seed liquor, it will
Seed liquor is inoculated in fluid nutrient medium according to the inoculum concentration of 5-10%, shake culture 5-7d, shaking speed 130-160rpm/
Min obtains culture solution；
S2, culture solution is subjected to centrifugal treating, isolated thallus and fermentation liquid is deposited in respectively and carry out cold lure under 0-8 DEG C of environment
Lead 20-40d；
S3, by after thallus and fermentation liquid that cold-induction is handled mix, carry out vacuum freeze drying, will freeze-drying mixture grinding
Cheng Fen obtains freeze-dried powder；
S4, freeze-dried powder is subjected to chloroform extraction, obtains primary extraction liquid, then filter paper filtering is carried out to primary extraction liquid, goes to clean
Matter；
S5, dialysis treatment is carried out to filtered primary extraction liquid, after the macromolecular for removing 500Da or more, obtains secondary extraction
Liquid；
S6, HPLC chromatogram purifying, isolated extract, i.e., non-carbohydrate extract are carried out to secondary extracting solution.
2. preparation method according to claim 1, which is characterized in that in step S1, the fluid nutrient medium used include with
The each component of lower mass percent: birch powder 5%, 10% glucose, 1.5% peptone, 0.2%KH2PO4, 0.2%MgSO4,
Remaining is water.
3. preparation method according to claim 1, which is characterized in that in step S3, the condition of vacuum freeze drying are as follows: will
After the mixture of thallus and fermentation liquid is placed in -20 DEG C of freezing 2-3h, then after freezing 4-8h under the conditions of being placed in -80 DEG C, by mixture
Taking-up is placed in vacuum freeze drier and is lyophilized, until obtaining freeze-drying mixture.
4. preparation method according to claim 1, which is characterized in that chloroform extraction methods are as follows: soxhlet's extraction method is used, with
Chloroform is dissolved and is extracted to freeze-dried powder as solvent, and 10-12h is impregnated, and is flowed back after 6-8h, is obtained primary extraction liquid.
5. preparation method according to claim 1, which is characterized in that in S6 step, the chromatographic column used is Hypersil
GOLD C18 column, column temperature are 28-32 DEG C.
6. preparation method according to claim 5, which is characterized in that in S6 step, using acetonitrile-water eluent system, water
With the volume ratio of acetonitrile are as follows: 1:(6-10), the flow velocity of secondary extracting solution is 0.2ml/min.
7. a kind of non-carbohydrate extract using Inonotus obliquus prepared by preparation method described in claim 1.
8. the non-carbohydrate extract of Inonotus obliquus as claimed in claim 7 prepare fungicide, the application in antibacterial medicines.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116584505A (en) * | 2023-04-23 | 2023-08-15 | 南京农丰生物科技有限公司 | Anti-staphylococcus aureus composite preparation and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040191340A1 (en) * | 2003-03-25 | 2004-09-30 | Council Of Scientific And Industrial Research | One step process for preparing antibacterial and antioxidant fraction from seabuckthorn parts (Hippophae rhamnoides L.) |
| CN109337953A (en) * | 2018-11-01 | 2019-02-15 | 黑龙江中医药大学 | A kind of inonotus obliquus D extract and preparation method thereof and detection method |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040191340A1 (en) * | 2003-03-25 | 2004-09-30 | Council Of Scientific And Industrial Research | One step process for preparing antibacterial and antioxidant fraction from seabuckthorn parts (Hippophae rhamnoides L.) |
| CN109337953A (en) * | 2018-11-01 | 2019-02-15 | 黑龙江中医药大学 | A kind of inonotus obliquus D extract and preparation method thereof and detection method |
Non-Patent Citations (1)
| Title |
|---|
| 师苑等: "桦褐孔菌不同提取物提取工艺及生物活性研究", 《广州化工》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116584505A (en) * | 2023-04-23 | 2023-08-15 | 南京农丰生物科技有限公司 | Anti-staphylococcus aureus composite preparation and preparation method and application thereof |
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