Method for extracting product rich in peroxy-ergosterol from lucid ganoderma

Abstract

The invention discloses a method for extracting a product rich in peroxy-ergosterol from lucid ganoderma, which comprises the following steps: crushing lucid ganoderma to obtain lucid ganoderma fine powder, spraying water into the lucid ganoderma fine powder, extruding and granulating, and drying to obtain lucid ganoderma fine powder particles; ethanol is used as an entrainer, the material-liquid ratio is (1: 1)-(1: 3) kg / L, the pressure of an extraction kettle is 25-35 MPa, and the temperature is 40-65 DEG C; the pressure of the separation kettle I is 10-12MPa, and the temperature is 50-60 DEG C; the pressure of the separation kettle II is 5-8 MPa, and the temperature is 50-60 DEG C; the flow rate is 300-800L / min, and the extraction time is 4-6 hours. According to the method disclosed by the invention, a product rich in peroxy-ergosterol, ergosterol and ganoderma triterpenes at the same time is obtained, and the safety, effectiveness and quality controllability of the product are considered while the effective active ingredients of ganoderma are enriched.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine processing, and particularly relates to a method for extracting a product rich in peroxyergosterol from ganoderma lucidum. Background Art

[0002] Natural products have a long history of medicinal use. Finding lead compounds from natural products and developing highly effective and low-toxic anti-tumor drugs is one of the important directions for new drug research and development. Ergosterol peroxide is a natural sterol 5α, 8α-epoxide that is found in a variety of edible fungi and has a variety of pharmacological activities, with anti-tumor and anti-inflammatory effects being the most prominent. Ergosterol is the main active ingredient in Ganoderma lucidum edible fungi and has good anti-tumor activity.

[0003] The invention is to extract a product rich in peroxyergosterol from ganoderma lucidum, especially containing ergosterol and ganoderma triterpenoids at the same time, which has good anti-cancer effect, and there is no technical report on this technology at present. The invention has low production cost, is green and environmentally friendly, has high extraction rate, and is particularly suitable for industrial production. Summary of the invention

[0004] The object of the present invention is to provide a method suitable for mass production for extracting a product rich in peroxyergosterol from Ganoderma lucidum.

[0005] To solve the above problems, the technical solution adopted by the present invention is:

[0006] A method for extracting a product rich in peroxyergosterol from Ganoderma lucidum, the method comprising the following steps:

[0007] (1) Grinding the Ganoderma lucidum through a 80-200 mesh sieve to obtain Ganoderma lucidum powder;

[0008] (2) spraying water into the Ganoderma lucidum powder, with the mass ratio of the material to the liquid being 0.5% to 1.5%, extruding into granules, and drying at 60° C. to 80° C. to obtain Ganoderma lucidum powder granules;

[0009] (3) The Ganoderma lucidum powder particles are subjected to CO2 supercritical extraction: ethanol with a mass concentration of 95% to 100% is used as an entrainer, the solid-liquid ratio is 1:1 to 1:3 kg / L, the pressure of the extraction kettle is 25 MPa to 35 MPa, and the temperature is 40°C to 65°C; the pressure of the separation kettle I is 10 MPa to 12 MPa, and the temperature is 50°C to 60°C; the pressure of the separation kettle II is 5 MPa to 8 MPa, and the temperature is 50°C to 60°C; the flow rate is 300 to 800 L / min, and the extraction time is 4 to 6 h, to obtain a product rich in peroxyergosterol.

[0010] Preferably, in the step (1), the Ganoderma lucidum is crushed and passed through a 120-mesh sieve to obtain Ganoderma lucidum powder.

[0011] Preferably, in the step (2), the solid-to-liquid ratio of spraying water into the Ganoderma lucidum powder is 1.0%.

[0012] Preferably, the drying temperature is 70°C.

[0013] Preferably, in the CO2 supercritical extraction in step (3), ethanol with a mass concentration of 95% is used as the entrainer.

[0014] Preferably, the material-liquid ratio is 1:2 kg / L.

[0015] Preferably, the extraction kettle pressure is 30 MPa, the temperature is 55° C., and the extraction time is 5 h.

[0016] Preferably, the pressure of the separation kettle I is 11 MPa and the temperature is 55°C.

[0017] Preferably, the separation kettle II has a pressure of 7 MPa, a temperature of 55° C., and a flow rate of 700 L / min.

[0018] Preferably, the content of peroxyergosterol in the product obtained in step (3) is 0.5% to 1.5%, the content of ergosterol is 2.7% to 5%, the content of ganoderma triterpenoid is 5% to 9%, and the product yield is 1% to 2%.

[0019] The present invention particularly preferably uses an ethanol-to-liquid ratio of 1:2 kg / L, and under low-pressure and low-temperature CO2 supercritical extraction conditions, the heat-sensitive active ingredients are not destroyed, and peroxyergosterol, ergosterol and ganoderma triterpenes can be effectively extracted. The pressure of the separation kettle II is lower than 8 MPa, and there will be no solvent residue.

[0020] Compared with the prior art, the invention has the following beneficial effects: the invention uses ethanol as an entrainer to extract the ganoderma lucidum powder particles by supercritical CO2 to obtain a drug rich in peroxyergosterol, ergosterol and ganoderma lucidum triterpenoid components. The invention has a stable process, low energy consumption, and is suitable for mass production and processing. It enriches the effective active ingredients of ganoderma lucidum while taking into account safety, effectiveness and quality controllability. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the chromatogram of the peroxyergosterol reference substance in Example 1 of the present invention;

[0022] Figure 2 is a chromatogram of ergosterol peroxide in the product of Example 1 of the present invention;

[0023] Figure 3 This is the chromatogram of the ergosterol reference substance in Example 1 of the present invention;

[0024] Figure 4 is a chromatogram of ergosterol in the product of Example 1 of the present invention;

[0025] Figure 5 This is the chromatogram of the Ganoderma lucidum triterpenoid reference substance in Example 1 of the present invention;

[0026] Figure 6 It is the chromatogram of Ganoderma triterpenes in the product of Example 1 of the present invention. DETAILED DESCRIPTION

[0027] The technical solution of the present invention is further described in detail below in conjunction with several preferred embodiments. The following embodiments are implemented on the premise of the technical solution of the invention, and detailed implementation methods and specific operating processes are given, but the protection scope of the present invention is not limited to the following embodiments.

[0028] The raw material of Ganoderma lucidum used in the following examples is Ganoderma lucidum fruiting body purchased from Anhui Huangshan Yunle Ganoderma Lucidum Co., Ltd. The content of peroxyergosterol was 0.03%, the content of ergosterol was 0.15%, and the content of Ganoderma lucidum triterpenoid was 0.24%.

[0029] Example 1

[0030] (1) crushing the ganoderma lucidum through a 120-mesh sieve to obtain ganoderma lucidum powder;

[0031] (2) spraying water into the Ganoderma lucidum powder (the material-liquid mass ratio is 1.0%), extruding granules, and drying at 70° C. to obtain Ganoderma lucidum powder granules;

[0032] (3) The Ganoderma lucidum powder particles were subjected to CO2 supercritical extraction: 95% ethanol was used as an entrainer, the solid-liquid ratio was 1:2 kg / L, the extraction kettle pressure was 30 MPa, the temperature was 55°C; the separation kettle I pressure was 11 MPa, the temperature was 55°C; the separation kettle II pressure was 7 MPa, the temperature was 55°C; the flow rate was 700 L / min, and the extraction time was 5 h. The extraction obtained a yellow-brown paste, the content of peroxyergosterol was 1.0%, the content of ergosterol was 3%, the content of Ganoderma lucidum triterpenoid was 6%, and the product yield was 1%.

[0033] Example 2

[0034] (1) crushing the ganoderma lucidum through a 200-mesh sieve to obtain ganoderma lucidum powder;

[0035] (2) spraying water into the Ganoderma lucidum powder (the material-liquid mass ratio is 1.5%), extruding granules, and drying at 70° C. to obtain Ganoderma lucidum powder granules;

[0036] (3) The Ganoderma lucidum powder particles were subjected to CO2 supercritical extraction: 95% ethanol was used as an entrainer, the solid-liquid ratio was 1:3 kg / L, the extraction kettle pressure was 30 MPa, the temperature was 55°C; the separation kettle I pressure was 10 MPa, the temperature was 55°C; the separation kettle II pressure was 6 MPa, the temperature was 55°C; the flow rate was 700 L / min, and the extraction time was 5 h. The extraction obtained a yellow-brown paste, the content of peroxyergosterol was 0.8%, the content of ergosterol was 2.7%, the content of Ganoderma lucidum triterpenoid was 5.5%, and the product yield was 1.1%.

[0037] Example 3

[0038] (1) crushing the ganoderma lucidum through an 80-mesh sieve to obtain ganoderma lucidum powder;

[0039] (2) spraying water into the Ganoderma lucidum powder (the material-liquid mass ratio is 1.0%), extruding granules, and drying at 70° C. to obtain Ganoderma lucidum powder granules;

[0040] (3) The Ganoderma lucidum powder particles were subjected to CO2 supercritical extraction: 95% ethanol was used as an entrainer, the solid-liquid ratio was 1:2 kg / L, the extraction kettle pressure was 30 MPa, the temperature was 55°C; the separation kettle I pressure was 11 MPa, the temperature was 55°C; the separation kettle II pressure was 5 MPa, the temperature was 55°C; the flow rate was 700 L / min, and the extraction time was 5 h. The extraction obtained a yellow-brown paste, the content of peroxyergosterol was 0.85%, the content of ergosterol was 2.85%, the content of Ganoderma lucidum triterpenoid was 5.35%, and the product yield was 1.05%.

[0041] Comparative Example 1

[0042] (1) crushing the ganoderma lucidum through a 120-mesh sieve to obtain ganoderma lucidum powder;

[0043] (2) spraying water into the Ganoderma lucidum powder (the material-liquid mass ratio is 1.0%), extruding granules, and drying at 70° C. to obtain Ganoderma lucidum powder granules;

[0044] (3) The Ganoderma lucidum powder particles were subjected to CO2 supercritical extraction: 95% ethanol was used as an entrainer, the solid-liquid ratio was 1:5 kg / L, the extraction kettle pressure was 30 MPa, the temperature was 55°C; the separation kettle I pressure was 11 MPa, the temperature was 55°C; the separation kettle II pressure was 7 MPa, the temperature was 55°C; the flow rate was 700 L / min, and the extraction time was 5 h. The extraction obtained a yellow-brown paste, the content of peroxyergosterol was 0.45%, the content of ergosterol was 1.4%, the content of Ganoderma lucidum triterpenoid was 3.1%, and the product yield was 2%.

[0045] Comparative Example 2

[0046] (1) crushing the ganoderma lucidum through a 120-mesh sieve to obtain ganoderma lucidum powder;

[0047] (2) spraying water into the Ganoderma lucidum powder (the material-liquid mass ratio is 1.0%), extruding granules, and drying at 70° C. to obtain Ganoderma lucidum powder granules;

[0048] (3) The Ganoderma lucidum powder particles were subjected to CO2 supercritical extraction: 95% ethanol was used as an entrainer, the material-liquid ratio was 1:2 kg / L, the pressure of the extraction kettle was 30 MPa, and the temperature was 55°C; the pressure of the separation kettle I was 11 MPa, and the temperature was 55°C; the pressure of the separation kettle II was 10 MPa, and the temperature was 55°C; the flow rate was 700 L / min, and the extraction time was 5 h. The extraction obtained a yellow-brown liquid with residual ethanol, and then the ethanol was recovered by rotary evaporation for 2 hours to obtain a yellow-brown paste. The content of peroxyergosterol was 0.87%, the content of ergosterol was 2.6%, the content of Ganoderma lucidum triterpenoid was 5.2%, and the product yield was 1.15%.

[0049] The products prepared in the examples and comparative examples of the present invention were subjected to the content determination of the effective ingredients peroxyergosterol, ergosterol, and ganoderma triterpenoids, and the determination method was as follows:

[0050] 1. Ergosterol Peroxide

[0051] 1. Preparation of reference solution

[0052] Weigh an appropriate amount of ergosterol peroxide reference substance, accurately weigh it, and add methanol to make a solution containing 25 μg per 1 mL.

[0053] 2 Preparation of test solution

[0054] Weigh about 0.1 g of the sample, accurately weigh it, place it in a 10 mL volumetric flask, dilute it to the mark with methanol, shake it well, and you have it.

[0055] 3 Chromatographic reference conditions

[0056] Octadecylsilane bonded silica gel was used as filler; 100% methanol was used as mobile phase; the detector was ELSD; the drift tube temperature was 60°C, and the atomization temperature was 55°C.

[0057] 4 Determination

[0058] Accurately pipette 20 μL of reference solution and test solution respectively, inject into liquid chromatograph, and measure to obtain the result.

[0059] 2. Ergosterol

[0060] 1. Preparation of reference solution

[0061] Weigh 10 mg of ergosterol reference substance accurately, place in a 10 mL volumetric flask, add anhydrous ethanol to dissolve and dilute to the scale, shake well, and obtain.

[0062] 2 Preparation of test solution

[0063] Take about 1g of sample, weigh accurately, place in a 10mL volumetric flask, add 50% chloroform methanol solution to dissolve, make up to the scale, shake well, and you have it.

[0064] 3 Chromatographic reference conditions

[0065] Octadecylsilane bonded silica gel was used as filler; 100% methanol was used as mobile phase; flow rate was 1.0 mL / min; column temperature was 30°C; the detector was an ultraviolet detector; and the detection wavelength was 282 nm.

[0066] 4 Determination

[0067] Accurately pipette 10 μL of reference solution and test solution respectively, inject into liquid chromatograph, and measure to obtain the result.

[0068] 3. Ganoderma lucidum triterpenes

[0069] The test was carried out according to the method for determining the triterpenoid content of Ganoderma lucidum under the item "Ganoderma lucidum" in dietary supplements in the United States Pharmacopeia-National Formulary (USP40-NF35).

[0070] The content of triterpenoids in Ganoderma lucidum is calculated as the sum of the contents of ganoderic acid A, B, C2, D, F, G, H and ganoderic acid B, C, D.

Claims

1. A method for extracting a product rich in ergosterol peroxide from Ganoderma lucidum, characterized in that The method comprises the following steps: (1) Grinding the Ganoderma lucidum through a 80-200 mesh sieve to obtain Ganoderma lucidum powder; (2) spraying water into the Ganoderma lucidum powder with a solid-liquid ratio of 0.5% to 1.5%, extruding and granulating, and drying at 60° C. to 80° C. to obtain Ganoderma lucidum powder particles; (3) The Ganoderma lucidum powder particles are subjected to CO2 supercritical extraction: ethanol with a mass concentration of 95% to 100% is used as an entrainer, the solid-liquid ratio is 1:1 to 1:3 kg / L, the pressure of the extraction kettle is 25 MPa to 35 MPa, and the temperature is 40°C to 65°C; the pressure of the separation kettle I is 10 MPa to 12 MPa, and the temperature is 50°C to 60°C; the pressure of the separation kettle II is 5 MPa to 8 MPa, and the temperature is 50°C to 60°C; the flow rate is 300 to 800 L / min, and the extraction time is 4 to 6 h, to obtain a product rich in peroxyergosterol.

2. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that In the step (1), the ganoderma lucidum is crushed and passed through a 120-mesh sieve to obtain ganoderma lucidum powder.

3. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that In the step (2), the solid-liquid ratio used for spraying water into the ganoderma lucidum powder is 1.0%.

4. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that The drying temperature is 70°C.

5. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that In the CO2 supercritical extraction in step (3), ethanol with a mass concentration of 95% is used as the entrainer.

6. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that The material-liquid ratio is 1:2 kg / L.

7. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that The extraction kettle pressure is 30MPa, the temperature is 55°C, and the extraction time is 5h.

8. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that The pressure of the separation kettle I is 11MPa and the temperature is 55°C.

9. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that The separation kettle II has a pressure of 7 MPa, a temperature of 55°C, and a flow rate of 700 L / min.

10. The method for extracting a product rich in peroxyergosterol according to claim 1, characterized in that The product obtained in step (3) has a peroxyergosterol content of 0.5% to 1.5%, an ergosterol content of 2.7% to 5%, a ganoderma triterpenoid content of 5% to 9%, and a product yield of 1% to 2%.

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