CN114106213B - Preparation method of inonotus obliquus crude polysaccharide dry powder - Google Patents
Preparation method of inonotus obliquus crude polysaccharide dry powder Download PDFInfo
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- CN114106213B CN114106213B CN202111369848.8A CN202111369848A CN114106213B CN 114106213 B CN114106213 B CN 114106213B CN 202111369848 A CN202111369848 A CN 202111369848A CN 114106213 B CN114106213 B CN 114106213B
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
The invention discloses a preparation method of inonotus obliquus crude polysaccharide dry powder, which comprises the following specific steps: s1, preparing inonotus obliquus into fungus powder and degreasing; s2, adding water into the degreased fungus powder, decocting and centrifuging to obtain primary supernatant and primary residue, adding water into the primary residue, decocting and centrifuging to obtain secondary supernatant and secondary residue, and combining the primary supernatant and the secondary supernatant; s3, adding alkali liquor into the secondary residues, standing and centrifuging to obtain primary supernatant alkali liquor and primary alkali residues, neutralizing the primary supernatant alkali liquor, adding alkali liquor into the primary alkali residues, standing and centrifuging to obtain secondary supernatant alkali liquor, neutralizing the secondary supernatant alkali liquor, and combining the primary supernatant alkali liquor and the secondary supernatant alkali liquor which are neutralized to neutrality; s4, dialyzing the alkali extract by using a dialysis bag; and S5, concentrating the water extract and the alkali extract dialysate under reduced pressure, combining, freezing and drying to obtain crude polysaccharide dry powder. The invention utilizes the materials in the water extraction process, and the water extraction method and the alkali extraction method are matched, thereby improving the yield of the crude polysaccharide dry powder.
Description
Technical Field
The invention relates to the technical field of edible fungus extraction. More specifically, the invention relates to a preparation method of inonotus obliquus crude polysaccharide dry powder.
Background
The main active substance of the inonotus obliquus is polysaccharide, and the polysaccharide extract extracted from the inonotus obliquus sclerotia has the effects of obviously reducing blood sugar, blood fat, tumor growth and the like. However, the texture of the sclerotium of the inonotus obliquus is hard, the cell wall is difficult to break, the internal active ingredients are difficult to dissolve out, and the extraction efficiency of the polysaccharide is low.
Disclosure of Invention
It is an object of the present invention to address at least the above problems and to provide at least the advantages described hereinafter.
Still another object of the present invention is to provide a method for preparing a dried powder of inonotus obliquus crude polysaccharide, which can improve the extraction rate of the polysaccharide.
To achieve these objects and other advantages in accordance with the present invention, there is provided a method for preparing a dry powder of a crude polysaccharide of inonotus obliquus, comprising:
s1, crushing and sieving sclerotium of inonotus obliquus to prepare fungus powder, and degreasing the fungus powder to obtain degreased fungus powder;
s2, adding distilled water into the degreased fungus powder, boiling and centrifuging to obtain primary supernatant and primary residue, adding distilled water into the primary residue, boiling and centrifuging to obtain secondary supernatant and secondary residue, and combining the primary supernatant and the secondary supernatant to obtain water extract;
s3, adding alkali liquor into the secondary residues, standing and centrifuging to obtain primary supernatant alkali liquor and primary alkali residues, neutralizing the primary supernatant alkali liquor to be neutral, adding alkali liquor into the primary alkali residues, standing and centrifuging to obtain secondary supernatant alkali liquor, neutralizing the secondary supernatant alkali liquor to be neutral, and combining the neutralized primary supernatant alkali liquor and the neutralized secondary supernatant alkali liquor to obtain alkali extract;
s4, dialyzing the alkali extract by using a dialysis bag of 200Da to obtain alkali extract dialysate;
and S5, concentrating the water extract and the alkali extract dialysate under reduced pressure, combining, freezing and drying to obtain crude polysaccharide dry powder.
Preferably, the bacterial powder is degreased 2 times in step S1 with ethanol with a volume fraction of 95%.
Preferably, the volume ratio of the defatted bacterial powder to the distilled water in the step S2 is 1; the volume ratio of the primary residue to distilled water was 1.
Preferably, the alkali liquor is a 2% NaOH solution, the volume ratio of the secondary residue to the alkali liquor is 1.
Preferably, in step S4, monosaccharide is added to the alkaline extract dialysate to a sugar content equal to the total sugar content of the alkaline extract solution before dialysis.
Preferably, the alkali-extracted dialysate in step S5 is concentrated at 65 ℃ under reduced pressure to 3 times the volume of the defatted bacteria powder to obtain an alkali-extracted concentrated solution;
and (3) concentrating the water extract at 65 ℃ under reduced pressure until the volume of the defatted bacteria powder is 5 times that of the defatted bacteria powder to obtain water extract concentrated solution, mixing the alkali extract concentrated solution and the water body concentrated solution, pre-freezing the mixture in a refrigerator at-80 ℃ for not less than 2 hours, and freeze-drying the mixture in a freeze dryer to obtain the crude polysaccharide dry powder.
Preferably, the dialysis time in step S4 is not less than 3 days, and the water changing frequency is 3 times/d.
The invention at least comprises the following beneficial effects:
firstly, after the sclerotium of inonotus obliquus is made into powder and degreased, water extraction is firstly carried out to obtain water extract, the water extract is subsequently processed to obtain crude polysaccharide dry powder, meanwhile, residue generated in the water extraction process is subjected to alkali extraction to obtain alkali extract, the alkali extract is subsequently processed to obtain crude polysaccharide dry powder, and the two dry powders are combined to obtain the total crude polysaccharide dry powder extracted by the method.
Secondly, the alkali extraction dialysate obtained after alkali extraction liquid dialysis loses micromolecular solution, so that the internal balance of the alkali extraction dialysate is lost, a precipitate is formed, the precipitate cannot be redissolved in subsequent water adding, the redissolution effect of the extracted crude polysaccharide dry powder is poor, and the agglomeration phenomenon appears after redissolution.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
FIG. 1 is a sample of a dry powder of Inonotus obliquus crude polysaccharide prepared from an aqueous extract according to one embodiment of the present invention;
FIG. 2 is a state of the crude polysaccharide dry powder of FIG. 1 after reconstitution;
FIG. 3 is a sample of a dry powder of Fuscoporia obliqua crude polysaccharide prepared from the alkali extract according to one embodiment of the present invention;
fig. 4 is a state of the crude polysaccharide dry powder of fig. 3 after reconstitution.
Wherein, the preparation process of the crude polysaccharide dry powder in fig. 3 is the same as the procedure of example 2.
Detailed Description
The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.
Experimental materials: the wild air-dried inonotus obliquus sclerotium purchased from Changbai mountain for many years has the outer diameter length of not less than 20cm, hard and black appearance and yellow brown interior, is distributed with white channel-shaped hypha, and is identified as the inonotus obliquus sclerotium by professor Lihengtian of engineering research center of Ministry of edible and medicinal fungi education of Jilin agriculture university.
< example 1>
S1, weighing 50g of inonotus obliquus sclerotium, crushing, sieving with a 80-mesh sieve to obtain bacterial powder, immersing the bacterial powder in 1000mL of 95% ethanol by volume fraction, standing at room temperature for 24h, fully stirring for 3 times, centrifuging at the rotating speed of 5800rpm for 30min, taking out the precipitate, continuously adding 95% ethanol with the same volume into the precipitate, and repeating the operation to obtain degreased bacterial powder;
s2, adding 1500mL of distilled water into the degreased fungus powder, boiling the degreased fungus powder by using an induction cooker, then decocting the degreased fungus powder with soft fire for 2 hours, cooling the degreased fungus powder to room temperature, centrifuging the degreased fungus powder at the rotating speed of 5500rpm for 30 minutes to obtain primary supernatant and primary residue, adding distilled water with the same volume as that of the primary residue into the primary residue, repeatedly extracting the primary residue for one time to obtain secondary supernatant and secondary residue, and combining the primary supernatant and the secondary supernatant to obtain a water extract;
s3, adding 500mL of NaOH solution with the mass fraction of 2% into the secondary residues, quickly stirring uniformly, standing for 2h in a refrigerator at 4 ℃, centrifuging at the rotating speed of 5800rpm for 30min, collecting supernate to obtain primary supernatant alkali liquor, quickly adjusting the pH value to 7.0 by using concentrated hydrochloric acid, repeating the operation on the primary alkali residues to obtain secondary supernatant alkali liquor, quickly adjusting the pH value to 7.0 by using concentrated hydrochloric acid, and combining the neutralized primary supernatant alkali liquor and the neutralized secondary supernatant alkali liquor to obtain alkali extract;
s4, dialyzing the alkali extract by using a 200Da dialysis bag, wherein the dialysis time is not less than 3d, and the water changing frequency is 3 times/d, so as to obtain alkali extract dialysate;
s5, concentrating the alkali-extracted dialysate at 65 ℃ under reduced pressure until the volume of the degreased bacteria powder is 3 times that of the degreased bacteria powder, so as to obtain alkali-extracted concentrated liquid; concentrating the water extractive solution at 65 deg.C under reduced pressure to 5 times the volume of the defatted bacteria powder to obtain water extractive concentrated solution, mixing the alkali extractive concentrated solution and the water extractive concentrated solution, pre-freezing in refrigerator at-80 deg.C for no less than 2 hr, and freeze-drying in freeze drier to obtain crude polysaccharide dry powder.
< example 2>
The procedure is the same as in example 1, but in step S4, monosaccharide is added to the alkaline extract dialysate to a sugar content equal to the total sugar content of the alkaline extract before dialysis, wherein the monosaccharide may be mannose, glucose, or other sugar having a sweet taste.
< comparative example 1>
The polysaccharide extraction step of comparative example 1 is the same as S1 and S2 in example 1, the aqueous extract is concentrated in the same step S5, 95% ethanol in an amount of 3 to 4 times the volume of the aqueous extract is added to the aqueous extract while stirring, crude polysaccharide is precipitated, after centrifugation, a small amount of 95% ethanol is used for washing, centrifugation and repeated three times, the precipitate is collected and dried to obtain crude polysaccharide.
As a result:
the formula for calculating the polysaccharide extraction rate is as follows: polysaccharide extraction rate = polysaccharide extraction quality/fuscoporia obliqua fungus powder quality x 100%;
the yield of the crude polysaccharide dry powder is calculated according to the formula: dry powder yield = (the mass of the alkali extraction dry powder and the mass of the water extraction dry powder)/the mass of the inonotus obliquus bacterial powder is multiplied by 100%;
the formula for calculating the total sugar extraction rate is as follows: the total sugar extraction rate = total sugar extraction mass/fuscoporia obliqua fungus powder mass × 100%.
The following calculation is obtained through the calculation formula: the total sugar extraction rate of example 2 is 7.336%, the weight of the inonotus obliquus crude polysaccharide dry powder is 14.395g, and the polysaccharide extraction rate is 6.822%; the total polysaccharide extraction rate of comparative example 1 was 1.97%, the dry weight of the inonotus obliquus crude polysaccharide was 8.37g, and the polysaccharide extraction rate was 3.57%;
according to the data, the crude polysaccharide dry powder is prepared by matching water extraction and alkali extraction, so that the extraction rate of the crude polysaccharide dry powder is improved.
While embodiments of the invention have been described above, it is not intended to be limited to the details shown, described and illustrated herein, but is to be accorded the widest scope consistent with the principles and novel features herein disclosed, and to such extent that such modifications are readily available to those skilled in the art, and it is not intended to be limited to the details shown and described herein without departing from the general concept as defined by the appended claims and their equivalents.
Claims (6)
1. The preparation method of the inonotus obliquus crude polysaccharide dry powder is characterized by comprising the following steps:
s1, crushing and sieving inonotus obliquus to prepare fungus powder, and degreasing the fungus powder to obtain degreased fungus powder;
s2, adding distilled water into the degreased fungus powder, boiling and centrifuging to obtain primary supernatant and primary residue, adding distilled water into the primary residue, boiling and centrifuging to obtain secondary supernatant and secondary residue, and combining the primary supernatant and the secondary supernatant to obtain water extract;
s3, adding alkali liquor into the secondary residues, standing and centrifuging to obtain primary supernatant alkali liquor and primary alkali residues, neutralizing the primary supernatant alkali liquor to be neutral, adding alkali liquor into the primary alkali residues, standing and centrifuging to obtain secondary supernatant alkali liquor, neutralizing the secondary supernatant alkali liquor to be neutral, and combining the neutralized primary supernatant alkali liquor and the neutralized secondary supernatant alkali liquor to obtain alkali extract;
s4, dialyzing the alkali extract by using a dialysis bag of 200Da to obtain alkali extract dialysate;
s5, concentrating the water extract and the alkali extract dialysate under reduced pressure, combining, freezing and drying to obtain crude polysaccharide dry powder;
in step S4, monosaccharide is added to the alkali-extracted dialysate until the sugar content is equal to the total sugar content of the alkali-extracted liquid before dialysis.
2. The method for preparing a dry powder of a crude polysaccharide of inonotus obliquus according to claim 1, wherein the bacterial powder is defatted 2 times with ethanol having a volume fraction of 95% in step S1.
3. The method for preparing a dry powder of a crude polysaccharide of inonotus obliquus according to claim 1, wherein the volume ratio of the defatted bacteria powder to distilled water in step S2 is 1; the volume ratio of the primary residue to distilled water was 1.
4. The method for preparing the inonotus obliquus crude polysaccharide dry powder according to claim 1, wherein the alkali liquor is a 2% NaOH solution by mass fraction, the volume ratio of the secondary residue to the alkali liquor is 1.
5. The method for preparing a dried powder of a crude polysaccharide from Inonotus obliquus according to claim 1, wherein in step S5, the alkali-extracted dialysate is concentrated at 65 ℃ under reduced pressure to 3 times the volume of the defatted bacteria powder to obtain an alkali-extracted concentrate;
and (3) concentrating the water extract at 65 ℃ under reduced pressure until the volume of the defatted bacteria powder is 5 times that of the defatted bacteria powder to obtain a water extract concentrated solution, mixing the alkali extract concentrated solution and the water body concentrated solution, pre-freezing the mixture in a refrigerator at-80 ℃ for not less than 2 hours, and freeze-drying the mixture in a freeze dryer to obtain the crude polysaccharide dry powder.
6. The method for preparing a dry powder of a crude polysaccharide of inonotus obliquus according to claim 1, wherein the dialysis time in step S4 is not less than 3 days and the water exchange frequency is 3 times/d.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109134693A (en) * | 2018-11-19 | 2019-01-04 | 大兴安岭至臻尚品寒带生物技术有限公司 | A kind of preparation method of Fuscoporia obliqua polysaccharide |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109134693A (en) * | 2018-11-19 | 2019-01-04 | 大兴安岭至臻尚品寒带生物技术有限公司 | A kind of preparation method of Fuscoporia obliqua polysaccharide |
Non-Patent Citations (1)
| Title |
|---|
| 桦褐孔菌多糖的提取及功能的初步研究;杨超慧;《中国优秀博硕士学位论文全文数据库（硕士）医药卫生科技辑》;20100615(第6期);E057-103 * |
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