Preparation method of inonotuso bliquus polysaccharide extract

Abstract

The invention relates to the technical field of polysaccharide extraction, and provides a preparation method of an inonotuso bliquus polysaccharide extract, which comprises the following steps: S1, mixing inonotuso bliquus and a first extracting agent, extracting, and separating to obtain a filtered material; s2, mixing the filtered substance with a second extracting agent, extracting, and separating to obtain an extracting solution; s3, adding absolute ethyl alcohol into the extracting solution, standing, precipitating and centrifuging to obtain an inonotuso bliquus polysaccharide extract crude product, mixing the inonotuso bliquus polysaccharide extract crude product with a third extracting agent, extracting, taking a water phase, and concentrating to obtain an inonotuso bliquus polysaccharide extract; the first extracting agent comprises petroleum ether and isopropyl acetate; the second extracting agent comprises water; the third extracting agent comprises water, DL-lactic acid and dimethylformamide. According to the technical scheme, the problems of low extraction yield and low purity of the inonotuso bliquus polysaccharide extract in related technologies are solved.

Description

Technical Field

[0001] The invention relates to the technical field of polysaccharide extraction, and in particular to a method for preparing a chaga polysaccharide extract. Background Art

[0002] Chaga is a precious medicinal fungus, which contains rich chemical components, which play an important role in regulating human physiological functions, enhancing immunity, etc. Chaga contains rich triterpenoid compounds, polysaccharides, polyphenols and flavonoid compounds. Among them, polysaccharides, as one of the main components of Chaga, have significant immunomodulatory effects. They can activate immune cells, improve the body's resistance, help resist the invasion of diseases, and have great medicinal value. The purity of Chaga polysaccharide extract is crucial to maintaining its medicinal value. The traditional extraction method is hot water extraction. Although the operation is simple, the extraction efficiency is low, the polysaccharide loss is large, and the purity of the polysaccharide extract is low, which affects the quality and medicinal effect of Chaga polysaccharide extract. Therefore, adopting a preparation method of Chaga polysaccharide extract, it is a technical problem that needs to be solved urgently to obtain Chaga polysaccharide extract with high yield and high purity. Summary of the invention

[0003] The invention provides a method for preparing a chaga polysaccharide extract, which solves the problems of low extraction rate and low purity of the chaga polysaccharide extract in the related art.

[0004] The technical solution of the present invention is as follows: The present invention provides a method for preparing a chaga polysaccharide extract, comprising the following steps: S1, mixing Chaga with a first extractant, extracting, and separating to obtain a filtrate; S2, mixing the filtrate with a second extractant, extracting, and separating to obtain an extract; S3, adding anhydrous ethanol to the extract, standing and settling, centrifuging to obtain a crude Chaga polysaccharide extract, mixing the crude Chaga polysaccharide extract with a third extractant, extracting, taking an aqueous phase, and concentrating to obtain a Chaga polysaccharide extract; The first extractant includes petroleum ether and isopropyl acetate; The second extractant includes water; The third extractant includes water, DL-lactic acid and dimethylformamide.

[0005] As a further technical solution, the volume ratio of petroleum ether to isopropyl acetate is 4-9:1.

[0006] When the volume ratio of petroleum ether and isopropyl acetate is 4-9:1, the extraction rate and purity of Chaga polysaccharide extract can be further improved by reasonably adjusting the volume ratio of petroleum ether and isopropyl acetate.

[0007] As a further technical solution, the second extractant also includes lignin sulfonate and sodium bisulfite.

[0008] As a further technical solution, the second extractant comprises the following components in parts by weight: 80 parts of water, 18-25 parts of lignin sulfonate, 4-16 parts of sodium bisulfite.

[0009] When lignin sulfonate and sodium bisulfite are added to the second extractant, the joint extraction action of lignin sulfonate, sodium bisulfite and water helps the chaga polysaccharide extract to dissolve and dissolve, thereby further improving the extraction rate and purity of the chaga polysaccharide extract.

[0010] As a further technical solution, the weight ratio of the lignin sulfonate to sodium bisulfite is 2-4:1.

[0011] When the weight ratio of lignin sulfonate to sodium bisulfite is 2-4:1, the extraction rate and purity of Chaga polysaccharide extract can be further improved.

[0012] As a further technical solution, the lignin sulfonate is one or both of sodium lignin sulfonate and calcium lignin sulfonate.

[0013] As a further technical solution, the volume ratio of water, DL-lactic acid and dimethylformamide in the third extractant is 3~8:0.5:1.5.

[0014] As a further technical solution, in step S1, the chaga is cut and crushed chaga.

[0015] As a further technical solution, in g / mL, in step S1, the mass volume ratio of the chaga mushroom to the first extractant is 1 g: 20-40 mL.

[0016] As a further technical solution, in step S1, during the extraction, the temperature is 40-50° C. and the extraction time is 1-2 hours.

[0017] As a further technical solution, in step S2, the mass volume ratio of the filtrate to the second extractant is 1 g:50-70 mL, measured in g / mL.

[0018] As a further technical solution, in step S2, during the extraction, the temperature is 75-85°C and the extraction time is 1-2h.

[0019] As a further technical solution, in step S3, the mass volume ratio of the crude Chaga polysaccharide extract and the third extractant is 1 g:20-30 mL, measured in g / mL.

[0020] As a further technical solution, in step S3, during the static precipitation, the temperature is 5-10°C and the static time is 12-18h; during the centrifugation, the centrifugal speed is 3000-3500rpm and the centrifugal time is 10-15min.

[0021] As a further technical solution, in step S3, during the extraction, the temperature is 50-60° C. and the time is 1-2 h.

[0022] As a further technical solution, the volume ratio of the extract to anhydrous ethanol is 1:3-5.

[0023] The working principle and beneficial effects of the present invention are: In the present invention, the first extractant, the second extractant and the third extractant are used to extract the Chaga, and under the joint action of the three-step extraction, a Chaga polysaccharide extract with high yield and high purity can be obtained, wherein the first extractant includes petroleum ether and isopropyl acetate, and the joint action of petroleum ether and isopropyl acetate on the Chaga can reduce the interference of impurities in the subsequent extraction process of the Chaga polysaccharide extract, thereby significantly improving the yield and purity of the Chaga polysaccharide extract. In addition, when DL-lactic acid is present in the third extractant, the impurity content in the crude Chaga polysaccharide extract can be effectively removed, and the purity of the Chaga polysaccharide extract can also be significantly improved. DETAILED DESCRIPTION

[0024] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] Example 1 A method for preparing a chaga polysaccharide extract comprises the following steps: S1. Mix 10 g of the cut and crushed Chaga and the first extractant at a mass volume ratio of 1 g:20 mL, extract at 40° C. for 2 h, separate, and obtain a filtrate; S2, mixing the filtrate and the second extractant at a mass volume ratio of 1 g:50 mL, extracting at 75° C. for 2 h, separating, and obtaining an extract; S3, adding anhydrous ethanol to the extract, the volume of the extract and anhydrous ethanol is 1:3, standing at 5°C for 18 hours, centrifuging at a centrifugal speed of 3000 rpm for 15 minutes to obtain a crude Chaga polysaccharide extract, mixing the crude Chaga polysaccharide extract with the third extractant at a mass volume ratio of 1g:20mL, extracting at 50°C for 2 hours, taking the aqueous phase, and concentrating to obtain 1.133g of Chaga polysaccharide extract; The first extractant includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 7:3; The second extractant includes water; The third extractant includes water, DL-lactic acid and dimethylformamide, and the volume ratio of water, DL-lactic acid and dimethylformamide is 3:0.5:1.5.

[0026] Example 2 A method for preparing a chaga polysaccharide extract comprises the following steps: S1. Mix 10 g of the cut and crushed Chaga and the first extractant at a mass volume ratio of 1 g:30 mL, extract at 45° C. for 1.5 h, separate, and obtain a filtrate; S2, mixing the filtrate and the second extractant at a mass volume ratio of 1 g:60 mL, extracting at 80° C. for 1.5 h, separating, and obtaining an extract; S3, adding anhydrous ethanol to the extract, the volume of the extract and anhydrous ethanol is 1:4, standing at 8 ° C for 15 hours, centrifuging at a centrifugal speed of 3500 rpm for 10 minutes to obtain a crude Chaga polysaccharide extract, mixing the crude Chaga polysaccharide extract with the third extractant at a mass volume ratio of 1g:25mL, extracting at 55 ° C for 1.5 hours, taking the aqueous phase, and concentrating to obtain 1.246g of Chaga polysaccharide extract; The first extractant includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 7:3; The second extractant includes water; The third extractant includes water, DL-lactic acid and dimethylformamide, and the volume ratio of water, DL-lactic acid and dimethylformamide is 6:0.5:1.5.

[0027] Example 3 A method for preparing a chaga polysaccharide extract comprises the following steps: S1. Mix 10 g of the cut and crushed Chaga and the first extractant at a mass volume ratio of 1 g:40 mL, extract at 50° C. for 1 h, separate, and obtain a filtrate; S2, mixing the filtrate and the second extractant at a mass volume ratio of 1 g:70 mL, extracting at 85° C. for 1 h, separating, and obtaining an extract; S3, adding anhydrous ethanol to the extract, the volume of the extract and anhydrous ethanol is 1:5, standing for precipitation at 10°C for 12 hours, centrifuging at a centrifugal speed of 3500 rpm for 10 minutes to obtain a crude Chaga polysaccharide extract, mixing the crude Chaga polysaccharide extract with the third extractant at a mass volume ratio of 1g:30mL, extracting at 60°C for 1 hour, taking the aqueous phase, and concentrating to obtain 1.338g of Chaga polysaccharide extract; The first extractant includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 7:3; The second extractant includes water; The third extractant includes water, DL-lactic acid and dimethylformamide, and the volume ratio of water, DL-lactic acid and dimethylformamide is 8:0.5:1.5.

[0028] Example 4 The only difference between this embodiment and embodiment 2 is that in this embodiment, the first extracting agent includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 14:1; wherein, the obtained Chaga polysaccharide extract is 1.283g.

[0029] Example 5 The only difference between this embodiment and embodiment 2 is that in this embodiment, the first extracting agent includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 4:1; wherein, the obtained Chaga polysaccharide extract is 1.364g.

[0030] Example 6 The only difference between this embodiment and embodiment 2 is that in this embodiment, the first extracting agent includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 9:1; wherein, the obtained Chaga polysaccharide extract is 1.388g.

[0031] Example 7 The difference between this embodiment and embodiment 6 is that in this embodiment, in step S2, the filtrate and the second extractant are mixed in a mass volume ratio of 1 g:60 mL, and the second extractant includes the following components in parts by weight: 80 parts of water, 18 parts of sodium lignin sulfonate, and 4 parts of sodium bisulfite; wherein the obtained Chaga polysaccharide extract is 1.508 g.

[0032] Example 8 The difference between this embodiment and embodiment 6 is that in this embodiment, in step S2, the filtrate and the second extractant are mixed in a mass volume ratio of 1 g:60 mL, and the second extractant includes the following components in parts by weight: 80 parts of water, 19 parts of sodium lignin sulfonate, and 11 parts of sodium bisulfite; wherein the obtained Chaga polysaccharide extract is 1.570 g.

[0033] Example 9 The difference between this embodiment and embodiment 6 is that in this embodiment, in step S2, the filtrate and the second extractant are mixed in a mass volume ratio of 1 g:60 mL, and the second extractant includes the following components in parts by weight: 80 parts of water, 25 parts of sodium lignin sulfonate, and 16 parts of sodium bisulfite; wherein the obtained Chaga polysaccharide extract is 1.621 g.

[0034] Example 10 The only difference between this example and Example 8 is that in this example, 25 parts of sodium lignin sulfonate and 5 parts of sodium bisulfite are added to the second extractant; wherein, 1.597 g of Chaga polysaccharide extract is obtained.

[0035] Embodiment 11 The only difference between this example and Example 8 is that in this example, 20 parts of sodium lignin sulfonate and 10 parts of sodium bisulfite are added to the second extractant; wherein, 1.640 g of Chaga polysaccharide extract is obtained.

[0036] Example 12 The only difference between this example and Example 8 is that in this example, 24 parts of sodium lignin sulfonate and 6 parts of sodium bisulfite are added to the second extractant; wherein, 1.652 g of Chaga polysaccharide extract is obtained.

[0037] Comparative Example 1 The only difference between this comparative example and Example 1 is that in this comparative example, the first extracting agent only includes petroleum ether; wherein the obtained Chaga polysaccharide extract is 0.962 g.

[0038] Comparative Example 2 The only difference between this comparative example and Example 1 is that in this comparative example, the first extracting agent only includes isopropyl acetate; wherein the obtained Chaga polysaccharide extract is 0.875 g.

[0039] Comparative Example 3 The only difference between this comparative example and Example 1 is that in this comparative example, DL-lactic acid is not added to the first extractant; wherein the obtained Chaga polysaccharide extract is 0.994 g.

[0040] The Chaga polysaccharide extracts prepared by the preparation methods of Examples 1 to 12 and Comparative Examples 1 to 3 were tested for polysaccharide purity in the Chaga polysaccharide extracts by the phenol-sulfuric acid method, and the yield was calculated according to Chaga polysaccharide extract yield (%) = Chaga polysaccharide extract mass (g) × purity / Chaga raw material (g) × 100%. The test results are shown in Table 1 below.

[0041] Table 1 Test results of yield and purity of Chaga polysaccharide extracts of Examples 1 to 12 and Comparative Examples 1 to 3

[0042] Compared with Comparative Examples 1 and 2, the yield of the Chaga polysaccharide extract and the purity of the polysaccharide in the Chaga polysaccharide extract of Example 1 are significantly improved, indicating that when the first extracting agent includes petroleum ether and isopropyl acetate, the yield and purity of the Chaga polysaccharide extract can be significantly improved through the combined action of petroleum ether and isopropyl acetate on Chaga. Compared with Comparative Example 3, the purity of the polysaccharide in the Chaga polysaccharide extract of Example 1 is significantly improved, indicating that the addition of DL-lactic acid in the third extracting agent can also significantly improve the purity of the Chaga polysaccharide extract.

[0043] Compared with Examples 2 and 4, the yield of the Chaga polysaccharide extract and the purity of the polysaccharide in the Chaga polysaccharide extract of Examples 5-6 were improved, indicating that when the volume ratio of petroleum ether to isopropyl acetate was 4-9:1, the extraction yield and purity of the Chaga polysaccharide extract could be further improved.

[0044] Compared with Example 6, the yield of Chaga polysaccharide extract and the purity of polysaccharide in Chaga polysaccharide extract of Examples 7 to 9 were improved, indicating that when the second extractant also includes lignin sulfonate and sodium bisulfite, the extraction yield and purity of Chaga polysaccharide extract can be further improved under the joint extraction of lignin sulfonate, sodium bisulfite and water. Compared with Examples 8 and 10, the yield of Chaga polysaccharide extract and the purity of polysaccharide in Chaga polysaccharide extract of Examples 11 to 12 were improved, indicating that when the weight ratio of lignin sulfonate to sodium bisulfite is 2 to 4:1, the extraction yield and purity of Chaga polysaccharide extract can be further improved.

[0045] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a chaga polysaccharide extract, characterized in that: The following steps are involved: S1, mixing Chaga with a first extractant, extracting, and separating to obtain a filtrate; S2, mixing the filtrate with a second extractant, extracting, and separating to obtain an extract; S3, adding anhydrous ethanol to the extract, standing and settling, centrifuging to obtain a crude Chaga polysaccharide extract, mixing the crude Chaga polysaccharide extract with a third extractant, extracting, taking an aqueous phase, and concentrating to obtain a Chaga polysaccharide extract; The first extractant includes petroleum ether and isopropyl acetate; The second extractant includes water; The third extractant includes water, DL-lactic acid and dimethylformamide.

2. The method for preparing a Chaga polysaccharide extract according to claim 1, characterized in that: The volume ratio of petroleum ether to isopropyl acetate is 4-9:

1.

3. The method for preparing a Chaga polysaccharide extract according to claim 1, characterized in that: The second extractant also includes lignin sulfonate and sodium bisulfite.

4. The method for preparing a Chaga polysaccharide extract according to claim 3, characterized in that: The second extractant comprises the following components in parts by weight: 80 parts of water, 18-25 parts of lignin sulfonate, 4-16 parts of sodium bisulfite.

5. The method for preparing a Chaga polysaccharide extract according to claim 4, characterized in that: The weight ratio of the lignin sulfonate to sodium bisulfite is 2-4:

1.

6. The method for preparing a Chaga polysaccharide extract according to claim 3, characterized in that: The lignin sulfonate is one or both of sodium lignin sulfonate and calcium lignin sulfonate.

7. The method for preparing a Chaga polysaccharide extract according to claim 1, characterized in that: The volume ratio of water, DL-lactic acid and dimethylformamide in the third extractant is 3-8:0.5:1.

5.

8. The method for preparing a Chaga polysaccharide extract according to claim 1, characterized in that: In terms of g / mL, in step S1, the mass volume ratio of the Chaga mushroom to the first extractant is 1 g: 20-40 mL.

9. The method for preparing a Chaga polysaccharide extract according to claim 1, characterized in that: In terms of g / mL, in step S2, the mass volume ratio of the filtrate to the second extractant is 1 g:50-70 mL.

10. The method for preparing a Chaga polysaccharide extract according to claim 1, characterized in that: In terms of g / mL, in step S3, the mass volume ratio of the crude Chaga polysaccharide extract to the third extractant is 1 g:20-30 mL; and the volume ratio of the extract to anhydrous ethanol is 1:3-5.

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