Method for extracting and separating tricholoma matsutake alcohol, cordycepic acid and tricholoma matsutake polysaccharide

Abstract

The invention discloses a method for extracting and separating tricholoma matsutake alcohol, cordycepic acid and tricholoma matsutake polysaccharide, and relates to the technical field of tricholoma matsutake leftover material processing and utilization. Comprising the following steps: S1, taking 100g-200g of tricholoma matsutake leftover materials, crushing, sieving, filling tricholoma matsutake powder into a supercritical extraction kettle, and adding 5%-20% of an entrainer to obtain a crude extract; s2, carrying out normal-phase silica gel column chromatography separation on the crude extract obtained in S1 to obtain a target component, and carrying out gel column chromatography separation and purification on the component to obtain a high-purity monomeric compound; s3, 100 g of residues in the extraction kettle in the S1 are subjected to water heating extraction. According to the method disclosed by the invention, efficient extraction and separation of effective components in the tricholoma matsutake are realized through multiple separation and purification technologies such as supercritical extraction, silica gel column chromatography and gel column chromatography, and the purity and the yield of the extract are improved; the method is suitable for utilization of waste such as tricholoma matsutake leftover materials, the production cost is reduced, and the resource utilization rate is increased.

Description

Technical Field

[0001] The invention relates to the technical field of processing and utilization of pine mushroom scraps, and in particular to a method for extracting and separating pine mushroom alcohol, cordycepic acid and pine mushroom polysaccharide. Background Art

[0002] Matsutake is an ecto-mycorrhizal fungus of pine, oak and other trees. It is rich in polysaccharides, proteins, peptides, matsutake alcohol and other ingredients, and has a wide range of pharmacological effects, including anti-tumor, anti-virus, anti-diabetes and cardiovascular disease, anti-oxidation, liver protection, anti-aging, anti-radiation, anti-mutation, anti-bacterial, promoting gastrointestinal motility and activating the immune system. Matsutake products are mainly primary processed products such as fresh products, dried products and quick-frozen products, and there are fewer deep-processed products. For example, Yunnan's exported matsutake products mainly include fresh matsutake, refrigerated matsutake, frozen matsutake and salted matsutake.

[0003] Through investigations in Gongbujiangda, Tibet, Ganzi, Sichuan, and other places, it was found that the development of matsutake products is mostly primary processing methods such as fresh matsutake slices, drying or freeze-drying. The remaining matsutake scraps after slicing account for about 10-15% of the total weight of matsutake. This part of the raw materials is mostly discarded or fermented into organic fertilizer, which is an extreme waste of matsutake resources. At present, the existing technical data show that there is a lack of comparative research on the extraction efficiency and purification quality of matsutake active ingredients, and research on the recycling of scraps from primary processing of matsutake is even rarer. With the continued popularity of matsutake resources and the increasingly scarce wild resources, the comprehensive utilization and development of matsutake resources is of great significance. Expanding the development of matsutake scraps, using modern new extraction technology and composite extraction methods to extract active ingredients such as matsutake polysaccharides, matsutake polypeptides, matsutake alcohol, and cordycepic acid, improving the yield of active ingredients, and developing high-purity biological preparations can greatly increase the economic added value of matsutake.

[0004] Matsutake is widely used in food, medicine and cosmetics, with a wide variety of categories. Most products do not use ingredients extracted and purified from Matsutake, so there is great potential in developing products with single active ingredients such as Matsutake polysaccharides, Matsutake polypeptides, cordycepic acid and Matsutake alcohol.

[0005] After searching, the application scheme of Chinese patent application No. 201510735216.7 discloses a method for extracting polysaccharides from pine mushrooms, which method comprises the following steps: (1) extracting pine mushrooms with a hydrochloric acid solution having a mass fraction of 0.25-0.35% at 30-40°C for 1-3 hours to obtain an extract; (2) adding alkali to the extract to adjust the pH value to 7.5-8.0, and then removing insoluble matter by solid-liquid separation to obtain a clear aqueous solution A; (3) adding acid to the aqueous solution A to adjust the pH value to 7.5-8.0; and H value to 4.0-4.5, then solid-liquid separation to remove insoluble matter, to obtain a clear aqueous solution B; (4) adding the aqueous solution B to a macroporous adsorption resin column, first washing the macroporous adsorption resin with water to remove unadsorbed impurities; then washing the macroporous adsorption resin with a volume fraction of 15-20% ethanol solution to remove impurities with higher polarity; then washing the macroporous adsorption resin with a volume fraction of 65-70% ethanol solution as a desorption liquid, collecting the washed desorption liquid, and concentrating and drying under reduced pressure to obtain a matsutake extract. The methods in the above literature have the following shortcomings: poor environmental protection, and large resource consumption, which need to be improved. Summary of the invention

[0006] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a method for extracting and separating matsutake alcohol, cordycepic acid and matsutake polysaccharide.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide, comprising the following steps:

[0009] S1: 100g-200g of leftover matsutake materials are crushed and sieved, and the matsutake powder is put into a supercritical extraction kettle, and an entrainer is added at 5%-20% to obtain a crude extract;

[0010] S2: subjecting the crude extract obtained in S1 to normal phase silica gel column chromatography to obtain a target component, and subjecting the component to gel column chromatography separation and purification to obtain a high-purity monomer compound;

[0011] S3: 100 g of the residue in the extraction kettle of S1 was extracted by heating with water at a solid-liquid ratio of 1:5-1:15 (g / g), filtered, and concentrated to obtain a pine mushroom extract;

[0012] S4: taking the Tricholoma matsutake extract prepared in S3 and subjecting it to alcohol precipitation treatment with a mixed solvent, wherein the ratio of the Tricholoma matsutake extract to the mixed solvent is 1:2-1:5 (g / g);

[0013] S5: The supernatant after the alcohol precipitation in S4 is concentrated to obtain a refined extract, and the refined extract is added with a solution → crystallization → crystals are washed out → crude crystals are redissolved → recrystallization, and a high-purity crystalline substance is obtained through repeated recrystallization, separation and purification;

[0014] S6: structural identification of the crystals of S5;

[0015] S7: Weigh the precipitate after alcohol precipitation in S4, add distilled water to the precipitate at a solid-liquid ratio of 1:10-1:25, stir and dissolve in a constant temperature water bath at 85-95°C, let stand and then centrifuge to obtain a crude extract of Tricholoma matsutake polysaccharide;

[0016] S8: Take a certain amount of S7 Tricholoma matsutake polysaccharide crude extract, add Sevage reagent to the solution, seal it, shake it in a shaker for 30-60 minutes, pour it into a separatory funnel and let it stand for 30 minutes, remove the lower organic phase, repeat 3 to 5 times, then use a dialysis bag to dialyze it with running water to remove small molecular impurities, concentrate and freeze-dry it to obtain Tricholoma matsutake polysaccharide.

[0017] As a preferred embodiment of the present invention: the sieve mesh number in S1 is 100 mesh, the extraction temperature is 30-60°C, the extraction pressure is 20-40MPa, the temperature of separation kettle I is 30-45°C, the pressure of separation kettle I is 8-14MPa, the temperature of separation kettle II is 30-45°C, the pressure of separation kettle II is 4-10MPa, and the extraction time is 30-60min.

[0018] As a preferred embodiment of the present invention: in S3, the extraction temperature is 50-100° C., and the extraction time is 2-6 hours;

[0019] In S4, the ethanol concentration is 50%-85%, the alcohol precipitation temperature is -20°C-25°C, and the alcohol precipitation time is 4-24h;

[0020] In S8, dialysis is performed with flowing water for 72-96 hours using a dialysis bag with a cutoff of 3500 Da.

[0021] As a preferred embodiment of the present invention: the entrainer in S1 is one or more of ethanol, acetone and ethyl acetate.

[0022] As a preferred embodiment of the present invention: the elution solvent of the gel column of S2 is chloroform:methanol=1:4.

[0023] As a preferred embodiment of the present invention, the mixed solvent of S4 is ethanol:propylene glycol, and the ratio is 10:0-6:4.

[0024] As a preferred embodiment of the present invention: the Sevage reagent of S7 is chloroform: n-butanol = 4:1 (V / V), and the added amount is polysaccharide: Sevage = 2:1-4:1 (V / V).

[0025] As a preferred embodiment of the present invention: in the method, when the material is stirred, a stirring device is used, and the stirring device comprises:

[0026] A device body, wherein a mounting plate is mounted on the device body;

[0027] A hollow stirring frame, which is rotatably mounted on a mounting plate, and a circumferentially distributed output pipe is arranged at the bottom of the hollow stirring frame;

[0028] A feeding chamber, which is integrally arranged at the top of the hollow stirring frame, and is connected to the output pipe through the hollow stirring frame;

[0029] A driving motor, wherein the driving motor is mounted on a mounting plate;

[0030] A driving gear, the shaft of which is drivingly connected to the output end of the driving motor; a driven gear distributed circumferentially is arranged outside the charging chamber, and the driven gear meshes with the driving gear;

[0031] The output pipe is arranged at the bottom of the device body and a control valve is arranged on the output pipe.

[0032] As a preferred embodiment of the present invention: an elastic bag is arranged on the top of the output tube, a spherical head is fixed on the top of the elastic bag, a guide column is fixed on the outside of the spherical head, and the guide column is slidably connected to the side wall of the top of the output tube; the inside of the elastic bag is connected to the inside of the output tube;

[0033] An annular seat is installed inside the device body, and the bottom surface of the annular seat is a corrugated structure; the spherical head slides on the bottom surface of the annular seat.

[0034] As a preferred embodiment of the present invention: a circumferentially distributed heat transfer seat is arranged on the inner side of the bottom of the device body, an impeller is rotatably installed on the outer side of the heat transfer seat, and both the heat transfer seat and the impeller are made of materials with good heat transfer properties; an installation cavity is arranged at the bottom of the heat transfer seat, a heating seat is installed at the bottom of the device body, and a heating end of the heating seat is installed in the installation cavity;

[0035] The hollow stirring frame is provided with ball-head rods evenly distributed in a circumference, and the impeller is located on the movement path of the ball-head rods.

[0036] The beneficial effects of the present invention are:

[0037] 1. The present invention realizes efficient extraction and separation of effective ingredients in Tricholoma matsutake through multiple separation and purification technologies such as supercritical extraction, silica gel column chromatography, gel column chromatography, etc., thereby improving the purity and yield of the extract; the method is suitable for the utilization of waste such as Tricholoma matsutake scraps, reducing production costs and improving resource utilization.

[0038] 2. The present invention is provided with a stirring device, which can work based on a driving motor to drive the feeding chamber to rotate, and then use a hollow stirring frame to perform stirring processing. Materials are added through the feeding chamber, and the materials are output from the output pipe after passing through the hollow stirring frame, which can achieve more complete mixing.

[0039] 3. The present invention provides structures such as a spherical head, an elastic bag and an annular seat, so that the spherical head can slide on the bottom surface of the annular seat during the rotation of the hollow stirring frame. Since the bottom surface of the annular seat is a corrugated structure, the spherical head can be periodically squeezed, thereby facilitating the output of materials, and the movement of the structure can better promote stirring, thereby improving work efficiency.

[0040] 4. The present invention provides a heating seat, an impeller, a ball head rod and other structures, and can heat the material in the device body based on the operation of the heating seat and the heat transfer between the heat transfer seat and the impeller. When the hollow stirring frame rotates, the ball head rod can be used to drive the impeller to rotate, thereby improving the stirring effect and heating uniformity. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 A schematic diagram of the crystallization of cordycepic acid in a method for extracting and separating matsutake alcohol, cordycepic acid and pine mushroom polysaccharide proposed by the present invention;

[0042] Figure 2 A schematic diagram of the chemical formula of cordycepic acid in a method for extracting and separating matsutake alcohol, cordycepic acid and pine mushroom polysaccharide proposed by the present invention;

[0043] Figure 3 A schematic diagram of the chemical formula of Tricholoma matsutake alcohol in a method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide proposed by the present invention;

[0044] Figure 4 A method for extracting and separating pine mushroom alcohol, cordycepic acid and pine mushroom polysaccharide proposed by the present invention 1 HNMR spectrum;

[0045] Figure 5 A method for extracting and separating pine mushroom alcohol, cordycepic acid and pine mushroom polysaccharide proposed by the present invention 13 CNMR spectrum;

[0046] Figure 6 The HMBC spectrum of cordycepic acid in the method for extracting and separating matsutake alcohol, cordycepic acid and pine mushroom polysaccharide proposed by the present invention;

[0047] Figure 7 This is the ESIMS (+) spectrum of cordycepic acid in the method for extracting and separating matsutake alcohol, cordycepic acid and pine mushroom polysaccharide proposed by the present invention;

[0048] Figure 8 A schematic diagram of the structure of a stirring device for a method of extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide proposed by the present invention;

[0049] Fig. 9This is a schematic structural diagram of a cross-section of a stirring device for a method for extracting and separating matsutake alcohol, cordycepic acid and matsutake polysaccharide proposed by the present invention;

[0050] Fig.10 A schematic diagram of the structure of an annular seat and an impeller in a method for extracting and separating matsutake alcohol, cordycepic acid and matsutake polysaccharide proposed by the present invention;

[0051] Fig.11 A schematic structural diagram of a hollow stirring frame and an output tube cross-section of a method for extracting and separating matsutake alcohol, cordycepic acid and matsutake polysaccharide proposed by the present invention;

[0052] Fig.12 The present invention provides a flow chart of a method for extracting and separating matsutake alcohol, cordycepic acid and matsutake polysaccharide.

[0053] In the figure: 1 device body, 2 heating seat, 3 output pipe, 4 driving motor, 5 driving gear, 6 feeding chamber, 7 driven gear, 8 mounting plate, 9 elastic bag, 10 annular seat, 11 hollow stirring frame, 12 impeller, 13 heat transfer seat, 14 ball head rod, 15 output pipe, 16 spherical head, 17 guide column. DETAILED DESCRIPTION

[0054] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0055] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0056] Embodiment 1:

[0057] A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide, comprising the following steps:

[0058] S1: 100 g of leftover matsutake materials were crushed and sieved through 100 mesh, and the matsutake powder was put into a supercritical extraction kettle, the extraction temperature was 30°C, the extraction pressure was 20 MPa, the temperature of separation kettle I was 30°C, the pressure of separation kettle I was 8 MPa, the temperature of separation kettle II was 30°C, the pressure of separation kettle II was 4 MPa, the extraction time was 30 min, and an entrainer of 95% ethanol and 10% were added to obtain a crude extract;

[0059] S2: The crude extract obtained from S1 was subjected to normal phase silica gel column chromatography to obtain the target component, and the component was separated and purified by gel column chromatography to obtain a high-purity monomer compound, which was identified as Tricholoma matsutake alcohol by GC-MS analysis and comparison, with a purity of 95%;

[0060] S3: 50 g of the residue in the extraction kettle of S1 was extracted by heating with water, with a solid-liquid ratio of 1:5 (g / g), an extraction temperature of 50°C, an extraction time of 2 h, filtered, and concentrated to obtain a pine mushroom extract with a yield of 90%;

[0061] S4: taking the Tricholoma matsutake extract prepared in S3 and subjecting it to mixed solvent alcohol precipitation treatment, the ratio of Tricholoma matsutake extract to mixed solvent is 1:2 (g / g), the ethanol concentration is 50%, the alcohol precipitation temperature is -20°C, and the alcohol precipitation time is 4h;

[0062] S5: The supernatant after the alcohol precipitation in S4 is concentrated to obtain a refined extract, and the refined extract is added with a solution → crystallization → crystals are washed out → crude crystals are redissolved → recrystallization, and a high-purity crystalline substance is obtained through repeated recrystallization, separation and purification;

[0063] S6: The crystals of S5 were structurally identified as cordycepic acid with a purity of 95%;

[0064] S7: Weigh the precipitate after alcohol precipitation in S4, add distilled water to the precipitate at a solid-liquid ratio of 1:10, stir and dissolve in a constant temperature water bath at 85-95°C, let stand and then centrifuge to obtain a crude extract of Tricholoma matsutake polysaccharide.

[0065] S8: Take a certain amount of S7 Tricholoma matsutake polysaccharide crude extract, add Sevage reagent to the solution, seal it, shake it in a shaker for 30 minutes, pour it into a separatory funnel and let it stand for 30 minutes, remove the lower organic phase, repeat 3 times, then use a dialysis bag with a cutoff of 3500Da to dialyze it with running water for 72 hours to remove small molecular impurities, concentrate and freeze-dry it to obtain Tricholoma matsutake polysaccharide with a purity of 75.59%.

[0066] Embodiment 2:

[0067] A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide, comprising the following steps:

[0068] S1: 150 g of leftover matsutake materials were crushed and sieved through 100 mesh, and the matsutake powder was put into a supercritical extraction kettle, the extraction temperature was 45°C, the extraction pressure was 30 MPa, the temperature of separation kettle I was 40°C, the pressure of separation kettle I was 12 MPa, the temperature of separation kettle II was 40°C, the pressure of separation kettle II was 8 MPa, the extraction time was 45 min, and 15% of entrainer was added to obtain a crude extract;

[0069] S2: The crude extract obtained from S1 was subjected to normal phase silica gel column chromatography to obtain the target component, and the component was separated and purified by gel column chromatography to obtain a high-purity monomer compound, which was identified as Tricholoma matsutake alcohol by GC-MS analysis and comparison, with a purity of 96%;

[0070] S3: 100 g of the residue in the extraction kettle of S1 was extracted by solvent extraction, with a solid-liquid ratio of 1:10 (g / g), an extraction temperature of 85°C, an extraction time of 4 h, filtered, and concentrated to obtain a matsutake extract with a yield of 100%;

[0071] S4: taking the Tricholoma matsutake extract prepared in S3 and subjecting it to mixed solvent alcohol precipitation treatment, the ratio of Tricholoma matsutake extract to mixed solvent is 1:3 (g / g), the ethanol concentration is 75%, the alcohol precipitation temperature is -10°C, and the alcohol precipitation time is 12h;

[0072] S5: The supernatant after the alcohol precipitation in S4 is concentrated to obtain a refined extract, and the refined extract is added with a solution → crystallization → crystals are washed out → crude crystals are redissolved → recrystallization, and a high-purity crystalline substance is obtained through repeated recrystallization, separation and purification;

[0073] S6: The crystals of S5 were structurally identified as cordycepic acid with a purity of 96%;

[0074] S7: Weigh the precipitate after alcohol precipitation in S4, add distilled water to the precipitate at a solid-liquid ratio of 1:10, stir and dissolve in a constant temperature water bath at 85-95°C, let stand and then centrifuge to obtain a crude extract of Tricholoma matsutake polysaccharide.

[0075] S8: Take a certain amount of S7 Tricholoma matsutake polysaccharide crude extract, add Sevage reagent to the solution, seal it, shake it in a shaker for 45 minutes, pour it into a separatory funnel and let it stand for 30 minutes, remove the lower organic phase, repeat 4 times, then use a dialysis bag with a cutoff of 3500Da to dialyze it with running water for 84 hours to remove small molecular impurities, concentrate and freeze-dry to obtain Tricholoma matsutake polysaccharide with a purity of 78.65%.

[0076] Embodiment 3:

[0077] A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide, comprising the following steps:

[0078] S1: 200 g of leftover matsutake materials were crushed and sieved through 100 mesh, and the matsutake powder was put into a supercritical extraction kettle, the extraction temperature was 60°C, the extraction pressure was 40 MPa, the temperature of separation kettle I was 45°C, the pressure of separation kettle I was 14 MPa, the temperature of separation kettle II was 45°C, the pressure of separation kettle II was 10 MPa, the extraction time was 60 min, and an entrainer of 95% ethanol and 20% was added to obtain a crude extract;

[0079] S2: The crude extract obtained from S1 is subjected to normal phase silica gel column chromatography to obtain the target component, and the component is separated and purified by gel column chromatography to obtain a high-purity monomer compound, which is identified as Tricholoma matsutake alcohol by GC-MS analysis and comparison, with a purity of 95-98%;

[0080] S3: 150 g of the residue in the extraction kettle of S1 was extracted by solvent extraction, with a solid-liquid ratio of 1:15 (g / g), an extraction temperature of 90°C, an extraction time of 6 h, filtered, and concentrated to obtain a pine mushroom extract with a yield of 110%;

[0081] S4: taking the Tricholoma matsutake extract prepared in S3 and subjecting it to mixed solvent alcohol precipitation treatment, the ratio of Tricholoma matsutake extract to mixed solvent is 1:5 (g / g), the ethanol concentration is 85%, the alcohol precipitation temperature is -5°C, and the alcohol precipitation time is 24h;

[0082] S5: The supernatant after the alcohol precipitation in S4 is concentrated to obtain a refined extract, and the refined extract is added with a solution → crystallization → crystals are washed out → crude crystals are redissolved → recrystallization, and a high-purity crystalline substance is obtained through repeated recrystallization, separation and purification;

[0083] S6: The crystals of S5 were structurally identified as cordycepic acid with a purity of 98%;

[0084] S7: Weigh the precipitate after alcohol precipitation in S4, add distilled water to the precipitate at a solid-liquid ratio of 1:10, stir and dissolve in a constant temperature water bath at 85-95°C, let stand and then centrifuge to obtain a crude extract of Tricholoma matsutake polysaccharide;

[0085] S8: Take a certain amount of S7 Tricholoma matsutake polysaccharide crude extract, add Sevage reagent to the solution, seal it, shake it in a shaker for 60 minutes, pour it into a separatory funnel and let it stand for 30 minutes, remove the lower organic phase, repeat 5 times, then use a dialysis bag with a cutoff of 3500Da to dialyze it with running water for 96 hours to remove small molecular impurities, concentrate and freeze-dry to obtain Tricholoma matsutake polysaccharide with a purity of 80.25%.

[0086] Embodiment 4:

[0087] A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide, wherein a stirring device is used when stirring materials, and the stirring device comprises:

[0088] A device body 1, on which a mounting plate 8 is mounted;

[0089] A hollow stirring frame 11, the hollow stirring frame 11 is rotatably mounted on the mounting plate 8, and a circumferentially distributed output pipe 15 is provided at the bottom of the hollow stirring frame 11;

[0090] A feeding chamber 6, which is integrally arranged at the top of the hollow stirring frame 11, and the feeding chamber 6 is connected to the output pipe 15 through the hollow stirring frame 11;

[0091] A driving motor 4, wherein the driving motor 4 is mounted on a mounting plate 8;

[0092] A driving gear 5, the shaft of which is drivingly connected to the output end of the driving motor 4; a driven gear 7 distributed circumferentially is arranged on the outside of the feeding chamber 6, and the driven gear 7 is meshed with the driving gear 5;

[0093] An output pipe 3, which is arranged at the bottom of the device body 1 and is provided with a control valve;

[0094] By setting up a stirring device, it can work based on the driving motor 4 to drive the feeding chamber 6 to rotate, and then use the hollow stirring frame 11 to perform stirring processing. The material is added through the feeding chamber 6, and the material passes through the hollow stirring frame 11 and is output from the output pipe 15, which can achieve more complete mixing.

[0095] In order to better mix and load; Fig.10 , Fig.11 As shown, an elastic bag 9 is arranged on the top of the output tube 15, a spherical head 16 is fixed on the top of the elastic bag 9, a guide column 17 is fixed on the outside of the spherical head 16, and the guide column 17 is slidably connected to the top side wall of the output tube 15; the inside of the elastic bag 9 is connected to the inside of the output tube 15;

[0096] An annular seat 10 is installed inside the device body 1, and the bottom surface of the annular seat 10 is a corrugated structure; a spherical head 16 slides on the bottom surface of the annular seat 10;

[0097] By providing structures such as the spherical head 16, the elastic bag 9 and the annular seat 10, the spherical head 16 can slide on the bottom surface of the annular seat 10 during the rotation of the hollow stirring frame 11. Since the bottom surface of the annular seat 10 is a corrugated structure, the spherical head 16 can be periodically squeezed, thereby facilitating the output of materials, and the movement of the structure can better promote stirring, thereby improving work efficiency.

[0098] In order to facilitate uniform heating; Fig.10 As shown, a circumferentially distributed heat transfer seat 13 is provided on the inner side of the bottom of the device body 1, and an impeller 12 is rotatably installed on the outer side of the heat transfer seat 13. Both the heat transfer seat 13 and the impeller 12 are made of materials with good heat conductivity. An installation cavity is provided at the bottom of the heat transfer seat 13, and a heating seat 2 is installed at the bottom of the device body 1, and the heating end of the heating seat 2 is installed in the installation cavity.

[0099] To improve the stirring and heating effect; Fig.10 As shown, the hollow stirring frame 11 is equipped with ball-end rods 14 evenly distributed around the circumference, and the impeller 12 is located on the movement path of the ball-end rods 14;

[0100] By providing structures such as a heating seat 2, an impeller 12 and a ball head rod 14, the material in the device body 1 can be heated based on the operation of the heating seat 2 and the heat transfer between the heat transfer seat 13 and the impeller 12. When the hollow stirring frame 11 rotates, the ball head rod 14 can be used to drive the impeller 12 to rotate, thereby improving the stirring effect and heating uniformity.

[0101] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide, characterized in that: The following steps are involved: S1: 100g-200g of leftover matsutake materials are crushed and sieved, and the matsutake powder is put into a supercritical extraction kettle, and an entrainer is added at 5%-20% to obtain a crude extract; S2: subjecting the crude extract obtained in S1 to normal phase silica gel column chromatography to obtain a target component, and subjecting the component to gel column chromatography separation and purification to obtain a high-purity monomer compound; S3: 100 g of the residue in the extraction kettle of S1 was extracted by heating with water at a solid-liquid ratio of 1:5-1:15 (g / g), filtered, and concentrated to obtain a pine mushroom extract; S4: taking the Tricholoma matsutake extract prepared in S3 and subjecting it to alcohol precipitation treatment with a mixed solvent, wherein the ratio of the Tricholoma matsutake extract to the mixed solvent is 1:2-1:5 (g / g); S5: The supernatant after the alcohol precipitation in S4 is concentrated to obtain a refined extract, and the refined extract is added with a solution → crystallization → crystals are washed out → crude crystals are redissolved → recrystallization, and a high-purity crystalline substance is obtained through repeated recrystallization, separation and purification; S6: structural identification of the crystals of S5; S7: Weigh the precipitate after alcohol precipitation in S4, add distilled water to the precipitate at a solid-liquid ratio of 1:10-1:25, stir and dissolve in a constant temperature water bath at 85-95°C, let stand and then centrifuge to obtain a crude extract of Tricholoma matsutake polysaccharide; S8: Take a certain amount of S7 Tricholoma matsutake polysaccharide crude extract, add Sevage reagent to the solution, seal it, shake it in a shaker for 30-60 minutes, pour it into a separatory funnel and let it stand for 30 minutes, remove the lower organic phase, repeat 3 to 5 times, then use a dialysis bag to dialyze it with running water to remove small molecular impurities, concentrate and freeze-dry it to obtain Tricholoma matsutake polysaccharide.

2. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 1, characterized in that: The sieve mesh number in S1 is 100 mesh, the extraction temperature is 30-60°C, the extraction pressure is 20-40MPa, the temperature of separation kettle I is 30-45°C, the pressure of separation kettle I is 8-14MPa, the temperature of separation kettle II is 30-45°C, the pressure of separation kettle II is 4-10MPa, and the extraction time is 30-60min.

3. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 2, characterized in that: In S3, the extraction temperature is 50-100° C. and the extraction time is 2-6 hours; In S4, the ethanol concentration is 50%-85%, the alcohol precipitation temperature is -20°C-25°C, and the alcohol precipitation time is 4-24h; In S8, dialysis is performed with flowing water for 72-96 hours using a dialysis bag with a cutoff of 3500 Da.

4. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 3, characterized in that: The entrainer in S1 is one or more of ethanol, acetone and ethyl acetate.

5. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 4, characterized in that: The elution solvent of the S2 gel column is chloroform:methanol=1:

4.

6. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 5, characterized in that: The mixed solvent of S4 is ethanol:propylene glycol, with a ratio of 10:0-6:

4.

7. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 6, characterized in that: The Sevage reagent of S7 is chloroform: n-butanol = 4:1 (V / V), and the added amount is polysaccharide: Sevage = 2:1-4:1 (V / V).

8. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to any one of claims 1 to 7, characterized in that: In the method, when the material is stirred, a stirring device is used, and the stirring device comprises: A device body (1), wherein a mounting plate (8) is mounted on the device body (1); A hollow stirring frame (11), the hollow stirring frame (11) is rotatably mounted on the mounting plate (8), and a circumferentially distributed output pipe (15) is provided at the bottom of the hollow stirring frame (11); A feeding chamber (6), the feeding chamber (6) is integrally arranged at the top of the hollow stirring frame (11), and the feeding chamber (6) is connected to the output pipe (15) through the hollow stirring frame (11); A drive motor (4), wherein the drive motor (4) is mounted on a mounting plate (8); A driving gear (5), the shaft of which is drivingly connected to the output end of the driving motor (4); driven teeth (7) distributed circumferentially are arranged on the outside of the feeding chamber (6), and the driven teeth (7) are meshed with the driving gear (5); An output pipe (3) is arranged at the bottom of the device body (1), and a control valve is arranged on the output pipe (3).

9. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 8, characterized in that: The top of the output tube (15) is provided with an elastic bag (9), a spherical head (16) is fixed on the top of the elastic bag (9), a guide column (17) is fixed outside the spherical head (16), and the guide column (17) is connected to the top side wall of the output tube (15) by sliding up and down; the inside of the elastic bag (9) is connected to the inside of the output tube (15); An annular seat (10) is installed inside the device body (1), and the bottom surface of the annular seat (10) is a corrugated structure; the spherical head (16) slides on the bottom surface of the annular seat (10).

10. A method for extracting and separating Tricholoma matsutake alcohol, cordycepic acid and Tricholoma matsutake polysaccharide according to claim 9, characterized in that: A heat transfer seat (13) distributed in a circumferential manner is arranged on the inner side of the bottom of the device body (1), and an impeller (12) is rotatably mounted on the outer side of the heat transfer seat (13), and both the heat transfer seat (13) and the impeller (12) are made of a material with good heat transfer properties; an installation cavity is arranged at the bottom of the heat transfer seat (13), and a heating seat (2) is mounted at the bottom of the device body (1), and a heating end of the heating seat (2) is mounted in the installation cavity; The hollow stirring frame (11) is provided with ball-head rods (14) evenly distributed around the circumference, and the impeller (12) is located on the movement path of the ball-head rods (14).

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