A preparation method of Inonotus obliquus polysaccharide extract

Abstract

The present invention relates to the technical field of polysaccharide extraction, and provides a preparation method of an inonotus obliquus polysaccharide extract, comprising the following steps: S1. Mix inonotus obliquus with a first extractant, extract and separate to obtain a filtrate; S2. Mix the filtrate with a second extractant, extract and separate to obtain an extract; S3. Add absolute ethanol to the extract, allow it to stand for precipitation, centrifuge to obtain a crude product of the inonotus obliquus polysaccharide extract, mix the crude product of the inonotus obliquus polysaccharide extract with a third extractant, extract, take the aqueous phase and concentrate to obtain the inonotus obliquus polysaccharide extract; the first extractant comprises petroleum ether and isopropyl acetate; the second extractant comprises water; the third extractant comprises water, DL-lactic acid and dimethylformamide. Through the above technical solution, the problems of low extraction yield and low purity of the inonotus obliquus polysaccharide extract in the related art are solved.

Description

Technical Field

[0001] The present invention relates to the technical field of polysaccharide extraction, and specifically, to a preparation method of an Inonotus obliquus polysaccharide extract. Background Art

[0002] Inonotus obliquus is a precious medicinal fungus, which contains rich chemical components, and these components play an important role in regulating human physiological functions and enhancing immunity. Inonotus obliquus contains rich triterpenoids, polysaccharides, polyphenols and flavonoids. Among them, polysaccharide compounds, as one of the main components of Inonotus obliquus, have significant immunomodulatory effects. It can activate immune cells, improve the body's resistance, help resist the invasion of diseases, and has great medicinal value. The purity of the Inonotus obliquus polysaccharide extract is crucial for 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 the Inonotus obliquus polysaccharide extract. Therefore, adopting a preparation method of an Inonotus obliquus polysaccharide extract to obtain an Inonotus obliquus polysaccharide extract with a higher yield and higher purity is a technical problem that needs to be solved urgently at present. Summary of the Invention

[0003] The present invention provides a preparation method of an Inonotus obliquus polysaccharide extract, which solves the problems of low extraction yield and low purity of the Inonotus obliquus polysaccharide extract in the related art.

[0004] The technical solution of the present invention is as follows:

[0005] The present invention provides a preparation method of an Inonotus obliquus polysaccharide extract, comprising the following steps:

[0006] S1. Mix Inonotus obliquus with a first extractant, extract and separate to obtain a filtrate;

[0007] S2. Mix the filtrate with a second extractant, extract and separate to obtain an extract;

[0008] S3. Add absolute ethanol to the extract, let it stand for precipitation, centrifuge to obtain a crude product of Inonotus obliquus polysaccharide extract, mix the crude product of Inonotus obliquus polysaccharide extract with a third extractant, extract, take the aqueous phase, and concentrate to obtain an Inonotus obliquus polysaccharide extract;

[0009] The first extractant includes petroleum ether and isopropyl acetate;

[0010] The second extractant includes water;

[0011] The third extractant includes water, DL-lactic acid and dimethylformamide.

[0012] As a further technical solution, the volume ratio of the petroleum ether to the isopropyl acetate is 4-9:1.

[0013] When the volume ratio of the petroleum ether to the isopropyl acetate is 4-9:1, by reasonably regulating the volume ratio of the petroleum ether to the isopropyl acetate, the extraction yield and purity of the chaga polysaccharide extract can be further improved.

[0014] As a further technical solution, the second extractant further includes lignosulfonate and sodium bisulfite.

[0015] As a further technical solution, the second extractant includes the following components in parts by weight:

[0016] 80 parts of water, 18-25 parts of lignosulfonate, and 4-16 parts of sodium bisulfite.

[0017] When lignosulfonate and sodium bisulfite are added to the second extractant, with the combined extraction effect of lignosulfonate, sodium bisulfite and water, it helps the dissolution and solution of the chaga polysaccharide extract, thereby further improving the extraction yield and purity of the chaga polysaccharide extract.

[0018] As a further technical solution, the weight ratio of the lignosulfonate to the sodium bisulfite is 2-4:1.

[0019] When the weight ratio of the lignosulfonate to the sodium bisulfite is 2-4:1, the extraction yield and purity of the chaga polysaccharide extract can be further improved.

[0020] As a further technical solution, the lignosulfonate is one or both of sodium lignosulfonate and calcium lignosulfonate.

[0021] 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.

[0022] As a further technical solution, in step S1, the chaga is the chaga that has been cut and crushed.

[0023] As a further technical solution, in step S1, based on g / mL, the mass-volume ratio of the chaga to the first extractant is 1 g:20-40 mL.

[0024] As a further technical solution, in step S1, during the extraction, the temperature is 40-50 °C and the extraction time is 1-2 h.

[0025] As a further technical solution, in step S2, based on g / mL, the mass-volume ratio of the filter residue to the second extractant is 1 g:50-70 mL.

[0026] As a further technical solution, in step S2, during the extraction, the temperature is 75 - 85°C and the extraction time is 1 - 2 h.

[0027] As a further technical solution, in step S3, calculated in g / mL, the mass - to - volume ratio of the crude Inonotus obliquus polysaccharide extract and the third extractant is 1 g:20 - 30 mL.

[0028] As a further technical solution, in step S3, during the static precipitation, the temperature is 5 - 10°C and the static time is 12 - 18 h; during the centrifugation, the centrifugation speed is 3000 - 3500 rpm and the centrifugation time is 10 - 15 min.

[0029] As a further technical solution, in step S3, during the extraction, the temperature is 50 - 60°C and the time is 1 - 2 h.

[0030] As a further technical solution, the volume ratio of the extract and absolute ethanol is 1:3 - 5.

[0031] The working principle and beneficial effects of the present invention are as follows:

[0032] In the present invention, the Inonotus obliquus is extracted with the first extractant, the second extractant and the third extractant. Under the combined action of the three - step extraction, an Inonotus obliquus polysaccharide extract with a relatively high yield and high purity can be obtained. Among them, the first extractant includes petroleum ether and isopropyl acetate. Through the combined action of petroleum ether and isopropyl acetate on Inonotus obliquus, the interference of impurities in the subsequent extraction process of the Inonotus obliquus polysaccharide extract can be reduced, thereby significantly improving the yield and purity of the Inonotus obliquus polysaccharide extract. In addition, when DL - lactic acid is present in the third extractant, the impurity content in the crude Inonotus obliquus polysaccharide extract can be effectively removed, and the purity of the Inonotus obliquus polysaccharide extract can also be significantly improved. Detailed implementation manners

[0033] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts fall within the scope of the present invention.

[0034] Example 1

[0035] A preparation method of an Inonotus obliquus polysaccharide extract includes the following steps:

[0036] S1. Mix 10 g of cut and pulverized Inonotus obliquus with the first extractant at a mass - to - volume ratio of 1 g:20 mL, extract at 40°C for 2 h, and separate to obtain a filtrate;

[0037] S2. Mix the filter residue and the second extractant at a mass-to-volume ratio of 1 g:50 mL, extract at 75 °C for 2 h, and separate to obtain an extract.

[0038] S3. Add absolute ethanol to the extract. The volume ratio of the extract to absolute ethanol is 1:3. Let it stand for precipitation at 5 °C for 18 h, centrifuge at a centrifugal speed of 3000 rpm for 15 min to obtain a crude extract of Inonotus obliquus polysaccharide. Mix the crude extract of Inonotus obliquus polysaccharide and the third extractant at a mass-to-volume ratio of 1 g:20 mL, extract at 50 °C for 2 h, take the aqueous phase, and concentrate to obtain 1.133 g of Inonotus obliquus polysaccharide extract.

[0039] The first extractant includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 7:3.

[0040] The second extractant includes water.

[0041] 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.

[0042] Example 2

[0043] A method for preparing an Inonotus obliquus polysaccharide extract, comprising the following steps:

[0044] S1. Mix 10 g of cut and crushed Inonotus obliquus and the first extractant at a mass-to-volume ratio of 1 g:30 mL, extract at 45 °C for 1.5 h, and separate to obtain a filter residue.

[0045] S2. Mix the filter residue and the second extractant at a mass-to-volume ratio of 1 g:60 mL, extract at 80 °C for 1.5 h, and separate to obtain an extract.

[0046] S3. Add absolute ethanol to the extract. The volume ratio of the extract to absolute ethanol is 1:4. Let it stand for precipitation at 8 °C for 15 h, centrifuge at a centrifugal speed of 3500 rpm for 10 min to obtain a crude extract of Inonotus obliquus polysaccharide. Mix the crude extract of Inonotus obliquus polysaccharide and the third extractant at a mass-to-volume ratio of 1 g:25 mL, extract at 55 °C for 1.5 h, take the aqueous phase, and concentrate to obtain 1.246 g of Inonotus obliquus polysaccharide extract.

[0047] The first extractant includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 7:3.

[0048] The second extractant includes water.

[0049] The third extractant comprises water, DL-lactic acid and dimethylformamide, and the volume ratio of water, DL-lactic acid and dimethylformamide is 6:0.5:1.5.

[0050] Example 3

[0051] A method for preparing an Inonotus obliquus polysaccharide extract, comprising the following steps:

[0052] S1. Mix 10 g of cut and crushed Inonotus obliquus with a first extractant at a mass-to-volume ratio of 1 g:40 mL, extract at 50 °C for 1 h, and separate to obtain a filtrate;

[0053] S2. Mix the filtrate with a second extractant at a mass-to-volume ratio of 1 g:70 mL, extract at 85 °C for 1 h, and separate to obtain an extract;

[0054] S3. Add absolute ethanol to the extract, with the volume ratio of the extract to absolute ethanol being 1:5, let it stand for precipitation at 10 °C for 12 h, centrifuge at a centrifugal speed of 3500 rpm for 10 min to obtain a crude Inonotus obliquus polysaccharide extract. Mix the crude Inonotus obliquus polysaccharide extract with a third extractant at a mass-to-volume ratio of 1 g:30 mL, extract at 60 °C for 1 h, take the aqueous phase, concentrate, and obtain 1.338 g of Inonotus obliquus polysaccharide extract;

[0055] The first extractant comprises petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 7:3;

[0056] The second extractant comprises water;

[0057] The third extractant comprises water, DL-lactic acid and dimethylformamide, and the volume ratio of water, DL-lactic acid and dimethylformamide is 8:0.5:1.5.

[0058] Example 4

[0059] The difference between this example and Example 2 is only that in this example, the first extractant comprises petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 14:1; wherein, 1.283 g of Inonotus obliquus polysaccharide extract is obtained.

[0060] Example 5

[0061] The difference between this example and Example 2 is only that in this example, the first extractant comprises petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 4:1; wherein, 1.364 g of Inonotus obliquus polysaccharide extract is obtained.

[0062] Example 6

[0063] The difference between this example and Example 2 is only that, in this example, the first extractant includes petroleum ether and isopropyl acetate, and the volume ratio of petroleum ether to isopropyl acetate is 9:1; among them, the obtained Inonotus obliquus polysaccharide extract is 1.388 g.

[0064] Example 7

[0065] The difference between this example and Example 6 is only that, in this example, in step S2, the filter residue and the second extractant are mixed at 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 lignosulfonate, and 4 parts of sodium bisulfite; among them, the obtained Inonotus obliquus polysaccharide extract is 1.508 g.

[0066] Example 8

[0067] The difference between this example and Example 6 is only that, in this example, in step S2, the filter residue and the second extractant are mixed at 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 lignosulfonate, and 11 parts of sodium bisulfite; among them, the obtained Inonotus obliquus polysaccharide extract is 1.570 g.

[0068] Example 9

[0069] The difference between this example and Example 6 is only that, in this example, in step S2, the filter residue and the second extractant are mixed at 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 lignosulfonate, and 16 parts of sodium bisulfite; among them, the obtained Inonotus obliquus polysaccharide extract is 1.621 g.

[0070] Example 10

[0071] The difference between this example and Example 8 is only that, in this example, the amount of sodium lignosulfonate added to the second extractant is 25 parts, and the amount of sodium bisulfite added is 5 parts; among them, the obtained Inonotus obliquus polysaccharide extract is 1.597 g.

[0072] Example 11

[0073] The difference between this example and Example 8 is only that, in this example, the amount of sodium lignosulfonate added to the second extractant is 20 parts, and the amount of sodium bisulfite added is 10 parts; among them, the obtained Inonotus obliquus polysaccharide extract is 1.640 g.

[0074] Example 12

[0075] The difference between this example and Example 8 is only that, in this example, the amount of sodium lignosulfonate added to the second extractant is 24 parts, and the amount of sodium bisulfite added is 6 parts; among them, the obtained Inonotus obliquus polysaccharide extract is 1.652 g.

[0076] Comparative Example 1

[0077] The difference between this comparative example and Example 1 is only that in this comparative example, the first extractant only includes petroleum ether; among them, the obtained Inonotus obliquus polysaccharide extract is 0.962 g.

[0078] Comparative Example 2

[0079] The difference between this comparative example and Example 1 is only that in this comparative example, the first extractant only includes isopropyl acetate; among them, the obtained Inonotus obliquus polysaccharide extract is 0.875 g.

[0080] Comparative Example 3

[0081] The difference between this comparative example and Example 1 is only that in this comparative example, DL-lactic acid is not added to the first extractant; among them, the obtained Inonotus obliquus polysaccharide extract is 0.994 g.

[0082] The Inonotus obliquus polysaccharide extracts prepared by the preparation methods of Examples 1 to 12 and Comparative Examples 1 to 3 were tested for the purity of polysaccharides in the Inonotus obliquus polysaccharide extract by the phenol-sulfuric acid method, and the yield was calculated according to the formula: yield of Inonotus obliquus polysaccharide extract (%) = mass of Inonotus obliquus polysaccharide extract (g) × purity / mass of Inonotus obliquus raw material (g) × 100%. The test results are shown in Table 1 below.

[0083] Table 1 Test results of the yield and purity of Inonotus obliquus polysaccharide extracts in Examples 1 to 12 and Comparative Examples 1 to 3

[0084]

[0085] Compared with Comparative Examples 1 to 2, the yield of the Inonotus obliquus polysaccharide extract and the purity of polysaccharides in the Inonotus obliquus polysaccharide extract in Example 1 were significantly improved, indicating that when the first extractant includes petroleum ether and isopropyl acetate, through the combined action of petroleum ether and isopropyl acetate on Inonotus obliquus, the yield and purity of the Inonotus obliquus polysaccharide extract can be significantly improved. Compared with Comparative Example 3, the purity of polysaccharides in the Inonotus obliquus polysaccharide extract in Example ① was significantly improved, indicating that the addition of DL-lactic acid to the third extractant can also significantly improve the purity of the Inonotus obliquus polysaccharide extract.

[0086] Compared with Examples 2 and 4, the yield of the Inonotus obliquus polysaccharide extract and the purity of polysaccharides in the Inonotus obliquus polysaccharide extract in Examples 5 to 6 were improved, indicating that when the volume ratio of petroleum ether to isopropyl acetate is 4 to 9:1, the extraction yield and purity of the Inonotus obliquus polysaccharide extract can be further improved.

[0087] Compared with Example 6, the yields of the Inonotus obliquus polysaccharide extract and the purity of the polysaccharide in the Inonotus obliquus polysaccharide extract in Examples 7 to 9 are increased, indicating that when the second extractant further includes lignosulfonate and sodium bisulfite, the extraction yield and purity of the Inonotus obliquus polysaccharide extract can be further improved under the combined extraction of lignosulfonate, sodium bisulfite and water. Compared with Examples 8 and 10, the yields of the Inonotus obliquus polysaccharide extract and the purity of the polysaccharide in the Inonotus obliquus polysaccharide extract in Examples 11 to 12 are increased, indicating that when the weight ratio of lignosulfonate to sodium bisulfite is 2 to 4:1, the extraction yield and purity of the Inonotus obliquus polysaccharide extract can be further improved.

[0088] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A preparation method of a polysaccharide extract from Inonotus obliquus, characterized in that, It includes the following steps: S1. Mix chaga with a first extracting agent, extract and separate to obtain a filtrate; S2. Mix the filtrate with a second extracting agent, extract and separate to obtain an extract; S3. Add absolute ethanol to the extract, let it stand for precipitation, centrifuge to obtain a crude product of chaga polysaccharide extract, mix the crude product of chaga polysaccharide extract with a third extracting agent, extract, take the aqueous phase and concentrate to obtain a chaga polysaccharide extract; The first extracting agent includes petroleum ether and isopropyl acetate; The second extracting agent includes water; The third extracting agent includes water, DL-lactic acid and dimethylformamide.

2. The preparation method of an Inonotus obliquus polysaccharide extract according to claim 1, characterized in that, The volume ratio of the petroleum ether to the isopropyl acetate is 4 - 9:

1.

3. The preparation method of a chaga polysaccharide extract according to claim 1, characterized in that, The second extracting agent further includes lignosulfonate and sodium bisulfite.

4. The preparation method of an Inonotus obliquus polysaccharide extract according to claim 3, characterized in that, The second extracting agent includes the following components in parts by weight: 80 parts of water, 18 - 25 parts of lignosulfonate, 4 - 16 parts of sodium bisulfite.

5. The preparation method of a chaga polysaccharide extract according to claim 4, characterized in that, The weight ratio of the lignosulfonate to the sodium bisulfite is 2 - 4:

1.

6. The preparation method of a chaga polysaccharide extract according to claim 3, characterized in that, The lignosulfonate is one or both of sodium lignosulfonate and calcium lignosulfonate.

7. A method for preparing an Inonotus obliquus polysaccharide extract according to claim 1, characterized in that, The volume ratio of water, DL-lactic acid and dimethylformamide in the third extracting agent is 3 - 8:0.5:1.

5.

8. The preparation method of a chaga polysaccharide extract according to claim 1, characterized in that, In step S1, the mass-to-volume ratio of the chaga to the first extracting agent is 1 g:20 - 40 mL in terms of g / mL.

9. The preparation method of an Inonotus obliquus polysaccharide extract according to claim 1, characterized in that, In step S2, the mass-to-volume ratio of the filtrate to the second extracting agent is 1 g:50 - 70 mL in terms of g / mL.

10. The preparation method of the chaga polysaccharide extract according to claim 1, wherein, In step S3, the mass-to-volume ratio of the crude product of chaga polysaccharide extract to the third extracting agent is 1 g:20 - 30 mL in terms of g / mL; the volume ratio of the extract to the absolute ethanol is 1:3 - 5.

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