Efficient extraction method and application of ganoderma lucidum spore oil

Abstract

The invention belongs to the technical field of plant extraction, and particularly relates to an efficient extraction method and application of ganoderma lucidum spore oil. Pretreated ganoderma lucidum spore powder is used as a raw material and is mixed with modified aluminum oxide nanoparticles and ethoxylated soybean lecithin, then a 1-butyl-3-methylimidazolium hexafluorophosphate solution is added, and the ganoderma lucidum spore oil is obtained through ultrasonic treatment and extraction treatment. By utilizing the physical wall breaking and group combination effect of the modified aluminum oxide nanoparticles, the chemical dissolution effect of the 1-butyl-3-methylimidazolium hexafluorophosphate and the emulsification and solubilization effects of the ethoxylated soya bean lecithin and combining with the mechanical effect of ultrasonic waves, multiple synergistic effects are formed, the extraction efficiency of the ganoderma lucidum spore oil is remarkably improved, and the extraction efficiency of the ganoderma lucidum spore oil is improved. The spore wall can be ruptured in a short time, and grease can be quickly released.

Description

Technical Field

[0001] The invention belongs to the technical field of plant extraction, and particularly relates to a high-efficiency extraction method of ganoderma lucidum spore oil and application thereof. Background Art

[0002] Ganoderma lucidum spore oil is a fat-soluble component extracted from Ganoderma lucidum spores. It is rich in a variety of bioactive substances that are beneficial to human health, including triterpenoids, unsaturated fatty acids, fat-soluble vitamins, sterols, etc. Ganoderma lucidum spores are the seeds of Ganoderma lucidum and the essence of Ganoderma lucidum. Ganoderma lucidum spore oil has received more and more attention due to its high bioavailability and unique pharmacological activity. Studies have shown that Ganoderma lucidum spore oil has anti-tumor, anti-oxidation, immune system regulation and other effects, and is widely used in health products and pharmaceutical preparations.

[0003] Ganoderma lucidum spores are tiny particles that are ejected from the cap of mature Ganoderma lucidum. Their outer layer is protected by a double-layer hard spore wall made of chitin. This structure effectively protects the active substances in the spores from the influence of the external environment, but it also brings challenges to extraction. The outer shell of Ganoderma lucidum spores is hard and the composition is complex, so it is very difficult to extract spore oil directly from Ganoderma lucidum spores. The content of Ganoderma lucidum spore oil is relatively low, and traditional extraction methods such as organic solvent extraction and supercritical CO2 extraction have certain limitations: for example, although cold pressing and water enzyme methods can extract Ganoderma lucidum spore oil, the extraction rate is low, especially in the mechanical cold pressing method, the Ganoderma lucidum spores are not completely broken, resulting in the inability to fully release the oil; in addition, the traditional organic solvent extraction method may cause residual harmful substances in the product, affecting its safety; the biologically active substances in Ganoderma lucidum spore oil are sensitive to temperature, and excessively high temperatures will cause the degradation of the effective ingredients, affecting its efficacy; the active ingredients of Ganoderma lucidum spore oil are very susceptible to high temperature, light, and oxygen, especially in organic solvent extraction methods and some high-temperature processes, active ingredients such as triterpenoid compounds and ganoderic acid are prone to degradation, resulting in a decrease in the quality of the extracted product. Therefore, based on the above technical problems, it is extremely necessary to develop a Ganoderma lucidum spore oil extraction method that can effectively reduce the use of organic solvents and is efficient and environmentally friendly. Summary of the invention

[0004] In view of the defects of the prior art, the object of the present invention is to provide a method for efficiently extracting Ganoderma lucidum spore oil and its application.

[0005] The technical effect of the present invention is achieved by the following technical solution: a method for efficiently extracting ganoderma lucidum spore oil, the specific preparation steps are as follows:

[0006] S1: Grinding the spore powder of Ganoderma lucidum through an 80-mesh sieve and drying it to obtain pretreated spore powder of Ganoderma lucidum; uniformly mixing the modified alumina nanoparticles and the pretreated spore powder of Ganoderma lucidum, and then adding a 5-10wt% ethoxylated soybean lecithin solution, stirring and mixing to obtain a mixed solution;

[0007] S2: slowly adding the 1-butyl-3-methylimidazolium hexafluorophosphate solution to the mixed solution prepared in step S1, stirring at a speed of 500 rpm to mix evenly, and then stirring at a speed of 200-300 rpm, maintaining the temperature at 35° C., and continuously stirring for 20-30 minutes to obtain an extraction system;

[0008] S3: placing the extraction system in step S2 in an ultrasonic reactor, and after the ultrasonic treatment is completed, cooling the extract to room temperature, centrifuging, filtering with a 0.22 μm filter membrane, washing with ethanol, and vacuum drying at 30° C. for 24 h to obtain Ganoderma lucidum spore oil.

[0009] Preferably, the specific preparation steps of the modified aluminum oxide nanoparticles described in step S1 are as follows:

[0010] A1: dissolving 3-aminopropyltriethoxysilane in anhydrous ethanol, stirring and dissolving uniformly to obtain a 3-aminopropyltriethoxysilane solution with a concentration of 2wt%; suspending aluminum oxide nanoparticles in anhydrous ethanol, ultrasonically treating, centrifuging, washing with anhydrous ethanol, and calcining in an oven at 500-700°C for 2-4h to obtain activated aluminum oxide nanoparticles;

[0011] A2: slowly adding the activated alumina nanoparticles prepared in step A1 to the 3-aminopropyltriethoxysilane solution prepared in step A1, heating to 60-80° C. and stirring for 12-24 hours, centrifuging, and washing with ethanol to obtain pretreated alumina nanoparticles;

[0012] A3: The pretreated alumina nanoparticles prepared in step A2 and succinic anhydride are added to ethyl acetate and mixed, reacted for 4 to 6 hours, washed with ethanol, centrifuged, and vacuum dried at 60° C. to constant weight to obtain modified alumina nanoparticles.

[0013] Preferably, the ratio of the amount of activated alumina nanoparticles to the amount of 3-aminopropyltriethoxysilane solution used in step A2 is 1 g: 2-3 mL.

[0014] Preferably, the ratio of the amount of the pretreated aluminum oxide nanoparticles, succinic anhydride and ethyl acetate used in step A3 is 1 g: 0.2-0.5 g: 20-30 mL.

[0015] Preferably, the specific preparation steps of the ethoxylated soybean lecithin in step S1 are as follows:

[0016] B1: Add soybean lecithin to the ethanol solution, stir and mix evenly to obtain a soybean lecithin solution with a concentration of 10-20%; add the soybean lecithin solution to a reactor, add 0.5% sodium hydroxide, seal and introduce nitrogen to maintain an inert atmosphere, then slowly introduce ethylene oxide gas at a rate of 15-20 mL / h, control the temperature to 100° C., the pressure to 0.2-0.5 MPa, and react for 2-4 hours;

[0017] B2: After the reaction in step B1 is completed, cool to room temperature, add dilute hydrochloric acid to neutralize the sodium hydroxide, take out the reaction product, wash it repeatedly with deionized water and acetone for 3 times, and vacuum dry it at 40-60°C for 6-12h to obtain ethoxylated soybean lecithin.

[0018] Preferably, the amount of the modified aluminum oxide nanoparticles in step S1 is 1.5-3% of the mass of the pretreated Ganoderma lucidum spore powder; the ratio of the amount of the ethoxylated soybean lecithin solution to the pretreated Ganoderma lucidum spore powder is 3mL:1g.

[0019] Preferably, the 1-butyl-3-methylimidazolium hexafluorophosphate solution in step S2 is prepared by mixing 1-butyl-3-methylimidazolium hexafluorophosphate and deionized water in a volume ratio of 1:4; the amount of 1-butyl-3-methylimidazolium hexafluorophosphate solution used is 20% of the volume of the mixed solution.

[0020] Preferably, the ultrasonic treatment parameters in step S3 are set to a frequency of 30-40 kHz, a power of 350-450 W, a system temperature of 32-37° C. maintained during the reaction, a treatment time of 30-50 min, and low-speed stirring at 200-300 rpm during the treatment.

[0021] Preferably, another aspect of the present invention is to provide an application of Ganoderma lucidum spore oil, wherein the Ganoderma lucidum spore oil can be effectively used in cosmetics and food additives.

[0022] The beneficial effects of the present invention are as follows:

[0023] The present invention utilizes the strong dissolving ability of 1-butyl-3-methylimidazolium hexafluorophosphate to effectively dissolve complex organic matter and biological molecules, and is used in combination with ethoxylated soybean lecithin to reduce the surface tension of ganoderma lucidum spore oil, while enhancing the diffusion and dissolution efficiency of oil in the solution, and improving the extraction rate. In addition, the ionic liquid has a good dissolving effect on organic molecules in the spore wall, which helps to further promote the rupture of the spore wall and help release the oil and active ingredients therein; the aluminum oxide nanoparticles exert physical shear force and friction effects under the synergistic effect of ultrasound through their high hardness and surface activity, further causing the spore wall to denature and trigger its structural destruction, promoting the release of oil and helping the release of oil. In addition, the aluminum oxide nanoparticles can also absorb part of the ultrasonic energy to produce a local micro-heating effect, which will not cause the overall temperature to rise, avoid damage to the active ingredients of the oil, and effectively promote the diffusion of oil from the spores, so that it can be quickly dissolved and released. 1-Butyl-3-methylimidazolium hexafluorophosphate not only has a strong affinity with Ganoderma lucidum spore oil, but also can reduce the interfacial tension between oil and water, further promoting the diffusion of oil from the spore wall. In addition, the ionic liquid can effectively disperse aluminum oxide nanoparticles and prevent the particles from settling in the liquid phase, thereby ensuring that the nanoparticles remain active during the extraction process, further accelerating the rupture of the spore wall and the release of oil. Ethoxylated modified soybean lecithin has excellent solubilizing and emulsifying abilities. Under the action of ultrasound, it can effectively reduce the surface tension of spore oil and form a stable emulsified system. This emulsification effect not only helps the stable dissolution and dispersion of oil in the aqueous phase, but also synergizes with the high solubility of 1-butyl-3-methylimidazolium hexafluorophosphate to further reduce the aggregation and coagulation of oil caused by interfacial tension during the extraction process, ensuring the uniform distribution of oil in the extraction system.

[0024] The cavitation effect of ultrasound can generate local high-intensity shear force in the liquid, and combined with the physical effect of alumina nanoparticles, it can further enhance the mechanical wall-breaking effect during the extraction process. The micro-stirring effect generated by ultrasonic oscillation can help alumina nanoparticles, ionic liquids and lecithin to be evenly distributed in the system, ensuring that they are in full contact with the spore wall of Ganoderma lucidum, accelerating the release of oil, and at the same time, through the emulsification of lecithin, making the oil more soluble. The carboxyl-modified alumina nanoparticles have higher chemical activity and hydrophilicity due to the introduction of carboxyl (-COOH) functional groups on the surface. The carboxyl group can interact with the amino or hydroxyl groups in the spore wall of Ganoderma lucidum through hydrogen bonds, further increasing the contact area between the alumina nanoparticles and the spore wall of Ganoderma lucidum, accelerating the rupture of the spore wall, and coordinating the mechanical action of ultrasound to accelerate the release of oil. In addition, the carboxyl modification improves the dispersibility of the nanoparticles in the aqueous system, forming a more stable dispersion system with ethoxylated soybean lecithin and 1-butyl-3-methylimidazole hexafluorophosphate, and enhancing the emulsification and solubilization effects.

[0025] Specifically, the physical wall breaking and group binding of modified alumina nanoparticles, the chemical dissolution of 1-butyl-3-methylimidazole hexafluorophosphate, the emulsification and solubilization of ethoxylated soybean lecithin, combined with the mechanical action of ultrasound, form multiple synergistic effects, significantly improving the extraction efficiency of Ganoderma lucidum spore oil, and can break the spore wall in a short time and quickly release oil. At the same time, this low-temperature extraction method can effectively avoid the degradation of unsaturated fatty acids and other active ingredients in Ganoderma lucidum spore oil, maintain the high quality of the extract, and the emulsification of lecithin also helps stabilize the oil, further reducing the risk of oxidation or degradation of active ingredients. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings in the following description are only for the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0027] Figure 1 It is a graph showing the test results of the extraction rate of Ganoderma lucidum spore oil of Example 2 of the present invention and Comparative Examples 1 to 4;

[0028] Figure 2 is a graph showing the test results of grease stability of Example 2 of the present invention and Comparative Examples 2 to 4;

[0029] Figure 3 It is a graph showing the wall breaking efficiency test of Example 2 of the present invention and Comparative Example 1;

[0030] Figure 4 It is a graph showing the acid value determination results of Example 2 of the present invention and Comparative Examples 1 to 4;

[0031] Figure 5 It is a free radical scavenging efficiency test chart of Examples 1 to 4 of the present invention and Comparative Examples 1 to 4. DETAILED DESCRIPTION

[0032] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them; 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. It should be noted that, unless otherwise specified, the raw materials involved in the present invention are purchased through conventional commercial channels.

[0033] Example 1: A highly efficient extraction method for Ganoderma lucidum spore oil, the specific preparation steps are as follows:

[0034] S1: Grind the Ganoderma lucidum spore powder, sieve it through an 80-mesh sieve, and dry it to obtain pretreated Ganoderma lucidum spore powder; mix 0.45 g of modified alumina nanoparticles and 30 g of the pretreated Ganoderma lucidum spore powder, then add 90 mL of 5 wt% ethoxylated soybean lecithin solution, stir and mix to obtain a mixed solution;

[0035] S2: Slowly add 26 mL of a solution prepared by mixing 1-butyl-3-methylimidazolium hexafluorophosphate and deionized water in a volume ratio of 1:4 to the mixed solution prepared in step S1, stir at 500 rpm to mix evenly, and then stir at 200 rpm, maintaining the temperature at 35° C., and continue stirring for 30 minutes to obtain an extraction system;

[0036] S3: placing the extraction system in step S2 in an ultrasonic reactor, setting the parameters to a frequency of 30 kHz, a power of 350 W, maintaining the system temperature at 32° C. during the reaction, and a treatment time of 50 min. During the treatment, stirring was performed at a low speed of 200 rpm. After the ultrasonic treatment was completed, the extract was cooled to room temperature, centrifuged, filtered through a 0.22 μm filter membrane, washed with ethanol, and vacuum dried at 30° C. for 24 h to obtain Ganoderma lucidum spore oil;

[0037] The specific preparation steps of the modified aluminum oxide nanoparticles described in step S1 are as follows:

[0038] A1: dissolving 3-aminopropyltriethoxysilane in anhydrous ethanol, stirring and dissolving uniformly to obtain a 3-aminopropyltriethoxysilane solution with a concentration of 2wt%; suspending aluminum oxide nanoparticles in anhydrous ethanol, ultrasonically treating, centrifuging, washing with anhydrous ethanol, and calcining in an oven at 500°C for 4h to obtain activated aluminum oxide nanoparticles;

[0039] A2: Slowly add 10 g of activated alumina nanoparticles prepared in step A1 into 20 mL of 3-aminopropyltriethoxysilane solution prepared in step A1, heat to 60° C., stir and react for 24 h, centrifuge, and wash with ethanol to obtain pretreated alumina nanoparticles;

[0040] A3: 10 g of the pretreated alumina nanoparticles prepared in step A2 and 2 g of succinic anhydride were added to 200 mL of ethyl acetate and mixed, reacted for 4 h, washed with ethanol, centrifuged, and dried under vacuum at 60° C. to constant weight to obtain modified alumina nanoparticles;

[0041] The specific preparation steps of the ethoxylated soybean lecithin described in step S1 are as follows:

[0042] B1: Add soybean lecithin to the ethanol solution, stir and mix evenly to obtain a 10% soybean lecithin solution; add 200 mL of the soybean lecithin solution to the reactor, add 1 g of sodium hydroxide, seal and introduce nitrogen to maintain an inert atmosphere, then slowly introduce ethylene oxide gas at a rate of 15 mL / h, control the temperature to 100°C, the pressure to 0.5 MPa, and react for 4 hours;

[0043] B2: After the reaction in step B1 is completed, cool to room temperature, add dilute hydrochloric acid to neutralize the sodium hydroxide, take out the reaction product, wash it repeatedly with deionized water and acetone for 3 times, and vacuum dry it at 40°C for 12 hours to obtain ethoxylated soybean lecithin.

[0044] Example 2: A highly efficient extraction method for Ganoderma lucidum spore oil, the specific preparation steps are as follows:

[0045] S1: Grind the Ganoderma lucidum spore powder, sieve it through an 80-mesh sieve, and dry it to obtain pretreated Ganoderma lucidum spore powder; mix 0.8 g of modified alumina nanoparticles and 30 g of the pretreated Ganoderma lucidum spore powder, then add 90 mL of 10 wt % ethoxylated soybean lecithin solution, stir and mix to obtain a mixed solution;

[0046] S2: Slowly add 26 mL of a solution prepared by mixing 1-butyl-3-methylimidazolium hexafluorophosphate and deionized water in a volume ratio of 1:4 to the mixed solution prepared in step S1, stir at 500 rpm to mix evenly, and then stir at 300 rpm, maintain the temperature at 35° C., and continue stirring for 25 minutes to obtain an extraction system;

[0047] S3: placing the extraction system in step S2 in an ultrasonic reactor, setting the parameters to a frequency of 35 kHz, a power of 400 W, maintaining the system temperature at 37°C during the reaction, a treatment time of 40 min, stirring at a low speed of 300 rpm during the treatment, and cooling the extract to room temperature after the ultrasonic treatment, centrifuging, filtering with a 0.22 μm filter membrane, washing with ethanol, and vacuum drying at 30°C for 24 h to obtain Ganoderma lucidum spore oil;

[0048] The specific preparation steps of the modified aluminum oxide nanoparticles described in step S1 are as follows:

[0049] A1: dissolving 3-aminopropyltriethoxysilane in anhydrous ethanol, stirring and dissolving uniformly to obtain a 3-aminopropyltriethoxysilane solution with a concentration of 2wt%; suspending aluminum oxide nanoparticles in anhydrous ethanol, ultrasonically treating, centrifuging, washing with anhydrous ethanol, and calcining in an oven at 700°C for 3h to obtain activated aluminum oxide nanoparticles;

[0050] A2: Slowly add 10 g of activated alumina nanoparticles prepared in step A1 into 30 mL of 3-aminopropyltriethoxysilane solution prepared in step A1, heat to 80° C., stir and react for 16 h, centrifuge, and wash with ethanol to obtain pretreated alumina nanoparticles;

[0051] A3: 10 g of the pretreated alumina nanoparticles prepared in step A2 and 3 g of succinic anhydride were added to 300 mL of ethyl acetate and mixed. After reacting for 6 h, the mixture was washed with ethanol, centrifuged, and dried under vacuum at 60° C. to constant weight to obtain modified alumina nanoparticles.

[0052] The specific preparation steps of the ethoxylated soybean lecithin described in step S1 are as follows:

[0053] B1: Add soybean lecithin to the ethanol solution, stir and mix well to obtain a 20% soybean lecithin solution; add 200 mL of the soybean lecithin solution to the reactor, add 1 g of sodium hydroxide, seal and introduce nitrogen to maintain an inert atmosphere, then slowly introduce ethylene oxide gas at a rate of 20 mL / h, control the temperature to 100°C, the pressure to 0.2 MPa, and react for 3 hours;

[0054] B2: After the reaction in step B1 is completed, cool to room temperature, add dilute hydrochloric acid to neutralize the sodium hydroxide, take out the reaction product, wash it repeatedly with deionized water and acetone for 3 times, and vacuum dry it at 60°C for 8 hours to obtain ethoxylated soybean lecithin.

[0055] Example 3: A highly efficient extraction method for Ganoderma lucidum spore oil, the specific preparation steps are as follows:

[0056] S1: Grind the Ganoderma lucidum spore powder, sieve it through an 80-mesh sieve, and dry it to obtain pretreated Ganoderma lucidum spore powder; mix 0.7 g of modified alumina nanoparticles and 30 g of the pretreated Ganoderma lucidum spore powder, then add 90 mL of 8 wt % ethoxylated soybean lecithin solution, stir and mix to obtain a mixed solution;

[0057] S2: Slowly add 26 mL of a solution prepared by mixing 1-butyl-3-methylimidazolium hexafluorophosphate and deionized water in a volume ratio of 1:4 to the mixed solution prepared in step S1, stir at 500 rpm to mix evenly, and then stir at 260 rpm, maintain the temperature at 35° C., and continue stirring for 25 min to obtain an extraction system;

[0058] S3: placing the extraction system in step S2 in an ultrasonic reactor, setting the parameters to a frequency of 35 kHz, a power of 400 W, maintaining the system temperature at 35° C. during the reaction, and a treatment time of 40 min. During the treatment, stirring was performed at a low speed of 260 rpm. After the ultrasonic treatment was completed, the extract was cooled to room temperature, centrifuged, filtered through a 0.22 μm filter membrane, washed with ethanol, and vacuum dried at 30° C. for 24 h to obtain Ganoderma lucidum spore oil;

[0059] The specific preparation steps of the modified aluminum oxide nanoparticles described in step S1 are as follows:

[0060] A1: dissolving 3-aminopropyltriethoxysilane in anhydrous ethanol, stirring and dissolving uniformly to obtain a 3-aminopropyltriethoxysilane solution with a concentration of 2wt%; suspending aluminum oxide nanoparticles in anhydrous ethanol, ultrasonically treating, centrifuging, washing with anhydrous ethanol, and calcining in an oven at 600°C for 3h to obtain activated aluminum oxide nanoparticles;

[0061] A2: Slowly add 10 g of activated alumina nanoparticles prepared in step A1 into 28 mL of 3-aminopropyltriethoxysilane solution prepared in step A1, heat to 70° C., stir and react for 20 h, centrifuge, and wash with ethanol to obtain pretreated alumina nanoparticles;

[0062] A3: 10 g of the pretreated alumina nanoparticles prepared in step A2 and 4 g of succinic anhydride were added to 260 mL of ethyl acetate and mixed. After reacting for 5 h, the mixture was washed with ethanol, centrifuged, and dried under vacuum at 60° C. to constant weight to obtain modified alumina nanoparticles.

[0063] The specific preparation steps of the ethoxylated soybean lecithin described in step S1 are as follows:

[0064] B1: Add soybean lecithin to the ethanol solution, stir and mix well to obtain a 15% soybean lecithin solution; add 200 mL of the soybean lecithin solution to the reactor, add 1 g of sodium hydroxide, seal and introduce nitrogen to maintain an inert atmosphere, then slowly introduce ethylene oxide gas at a rate of 18 mL / h, control the temperature to 100°C, the pressure to 0.3 MPa, and react for 4 hours;

[0065] B2: After the reaction in step B1 is completed, cool to room temperature, add dilute hydrochloric acid to neutralize the sodium hydroxide, take out the reaction product, wash it repeatedly with deionized water and acetone for 3 times, and vacuum dry it at 50°C for 10 hours to obtain ethoxylated soybean lecithin.

[0066] Example 4: A highly efficient extraction method for Ganoderma lucidum spore oil, the specific preparation steps are as follows:

[0067] S1: Grind the Ganoderma lucidum spore powder, sieve it through an 80-mesh sieve, and dry it to obtain pretreated Ganoderma lucidum spore powder; mix 0.9 g of modified alumina nanoparticles and 30 g of the pretreated Ganoderma lucidum spore powder, then add 90 mL of 10 wt % ethoxylated soybean lecithin solution, stir and mix to obtain a mixed solution;

[0068] S2: Slowly add 26 mL of a solution prepared by mixing 1-butyl-3-methylimidazolium hexafluorophosphate and deionized water in a volume ratio of 1:4 to the mixed solution prepared in step S1, stir at 500 rpm to mix evenly, and then stir at 300 rpm, maintain the temperature at 35° C., and continue stirring for 20 minutes to obtain an extraction system;

[0069] S3: placing the extraction system in step S2 in an ultrasonic reactor, setting the parameters to a frequency of 40 kHz, a power of 450 W, maintaining the system temperature at 37° C. during the reaction, and treating for 30 min. stirring at a low speed of 300 rpm during the treatment. After the ultrasonic treatment is completed, cooling the extract to room temperature, centrifuging, filtering with a 0.22 μm filter membrane, washing with ethanol, and vacuum drying at 30° C. for 24 h to obtain Ganoderma lucidum spore oil;

[0070] The specific preparation steps of the modified aluminum oxide nanoparticles described in step S1 are as follows:

[0071] A1: dissolving 3-aminopropyltriethoxysilane in anhydrous ethanol, stirring and dissolving uniformly to obtain a 3-aminopropyltriethoxysilane solution with a concentration of 2wt%; suspending aluminum oxide nanoparticles in anhydrous ethanol, ultrasonically treating, centrifuging, washing with anhydrous ethanol, and calcining in an oven at 700°C for 2h to obtain activated aluminum oxide nanoparticles;

[0072] A2: Slowly add 10 g of activated alumina nanoparticles prepared in step A1 into 30 mL of 3-aminopropyltriethoxysilane solution prepared in step A1, heat to 80° C., stir and react for 12 h, centrifuge, and wash with ethanol to obtain pretreated alumina nanoparticles;

[0073] A3: 10 g of the pretreated alumina nanoparticles prepared in step A2 and 5 g of succinic anhydride were added to 300 mL of ethyl acetate and mixed, reacted for 6 h, washed with ethanol, centrifuged, and dried under vacuum at 60° C. to constant weight to obtain modified alumina nanoparticles;

[0074] The specific preparation steps of the ethoxylated soybean lecithin described in step S1 are as follows:

[0075] B1: Add soybean lecithin to the ethanol solution, stir and mix well to obtain a 20% soybean lecithin solution; add 200 mL of the soybean lecithin solution to the reactor, add 1 g of sodium hydroxide, seal and introduce nitrogen to maintain an inert atmosphere, then slowly introduce ethylene oxide gas at a rate of 20 mL / h, control the temperature to 100°C, the pressure to 0.2 MPa, and react for 4 hours;

[0076] B2: After the reaction in step B1 is completed, cool to room temperature, add dilute hydrochloric acid to neutralize the sodium hydroxide, take out the reaction product, wash it repeatedly with deionized water and acetone for 3 times, and vacuum dry it at 60°C for 6 hours to obtain ethoxylated soybean lecithin.

[0077] Comparative Example 1: The operation of Comparative Example 1 is substantially the same as that of Example 2, except that no modified alumina nanoparticles are added in Comparative Example 1.

[0078] Comparative Example 2: The operation of Comparative Example 2 is basically the same as that of Example 2, except that no ethoxylated soybean lecithin solution is added in Comparative Example 2.

[0079] Comparative Example 3: The operation of Comparative Example 3 is substantially the same as that of Example 2, except that in Comparative Example 3, a soybean lecithin solution is used instead of an ethoxylated soybean lecithin solution.

[0080] Comparative Example 4: The operation of Comparative Example 4 is substantially the same as that of Example 2, except that ethyl acetate is used as the solvent in Comparative Example 4 instead of 1-butyl-3-methylimidazole hexafluorophosphate solution.

[0081] Performance Test:

[0082] Oil extraction rate test: The Ganoderma lucidum spore oil extracted from Example 2 and Comparative Examples 1 to 4 was placed in a dry container, the oil weight was weighed using a precision balance, and the Ganoderma lucidum spore oil extraction rate was calculated (extraction rate / (%) = extracted spore oil weight / initial weight of Ganoderma lucidum spore powder). Three test groups were set for each test, and the results were taken as the average of three tests. The results are shown in FIG. Figure 1 shown.

[0083] Depend on Figure 1As a result, the spore oil extraction method of the present invention can effectively realize the efficient extraction of Ganoderma lucidum spore oil; from the results of Comparative Example 1 and Example 2, it can be seen that the main function of the modified aluminum oxide nanoparticles is to exert mechanical shear force and friction effect on the spore wall under the action of ultrasound through its high hardness and surface activity, thereby accelerating the rupture of the spore wall. The lack of modified aluminum oxide nanoparticles means that there is a lack of effective physical wall-breaking means in the system, and the chemical action of the ionic liquid and ethoxylated soybean lecithin is also difficult to act deeply into the spores. In addition, the lack of nanoparticles has limited synergistic effect of the cavitation effect of ultrasound and the dissolution effect of the ionic liquid on destroying the spore wall, so that the extraction process mainly relies on chemical action rather than physical action, resulting in a decrease in extraction efficiency; from the results of Comparative Example 2 and Example 2, it can be seen that the lack of ethoxylated soybean lecithin has no synergistic effect of the ionic liquid and the nanoparticles. The method cannot be used, and the oil cannot be effectively emulsified and solubilized, which makes it difficult to disperse and stabilize the oil, and the synergy is seriously insufficient, which leads to a significant decrease in the oil extraction rate; from the results of Comparative Example 3 and Example 2, it can be seen that the solubilization ability of unmodified soybean lecithin is far less than that of ethoxylated lecithin, and it is difficult to effectively reduce the surface tension of the oil. After soybean lecithin replaces ethoxylated soybean lecithin, the emulsification and solubilization abilities are significantly reduced, and the dispersibility of modified alumina nanoparticles depends on the synergistic effect of lecithin and ionic liquids. After using unmodified soybean lecithin, the dispersion efficiency of the particles is reduced, the physical wall breaking effect and the micro-heating effect are weakened, resulting in a significant decrease in the extraction efficiency; from the results of Comparative Example 4 and Example 2, it can be seen that after ethyl acetate replaces the ionic liquid, the solubility, interfacial tension and dispersibility of the nanoparticles are greatly weakened, the synergy of the entire system is seriously invalid, and the extraction efficiency is significantly reduced.

[0084] Oil emulsification stability test: The extraction system prepared in step S2 of Example 2 and Comparative Examples 2 to 4 was mixed with deionized water in a ratio of 1:10, mixed with a homogenizer for 5 minutes, and then 10 mL was taken into a test tube and sealed at room temperature. On the 7th, 14th and 30th days, whether stratification, sedimentation or oil phase precipitation occurred during the observation period was observed, and then centrifuged at 3000 rpm for 15 minutes. After centrifugation, the test tube was taken out, the separation height of the oil phase and the water phase was observed, and the emulsification index / (%) was calculated = the height of the emulsion layer in the emulsion after offline / the total height of the emulsion before centrifugation × 100%. The test was repeated three times, and the results were averaged. The emulsification index results are as follows: Figure 2 The emulsification results are shown in Table 1.

[0085] Table 1. Emulsification stability results of the extraction system

[0086]

[0087] Depend on Figure 2As can be seen from the results of Table 1, the extraction system prepared by the present invention has excellent emulsification stability and also exhibits excellent stability during the entire storage period; from the results of Comparative Example 2 and Example 2, it can be seen that the absence of ethoxylated soybean lecithin reduces the solubilization and emulsification effects of the system, and the dispersibility of oil in the aqueous phase deteriorates, resulting in instability of the emulsified system during long-term storage, which in turn causes the stratification phenomenon to become obvious; from the results of Comparative Example 2 and Example 3, it can be seen that the hydrophilicity and solubilization ability of unmodified soybean lecithin are not as good as those of ethoxylated soybean lecithin, and the lack of synergy causes the emulsion system to gradually lose stability during long-term storage, which in turn causes the oil to aggregate and form stratification; from the results of Comparative Example 4 and Example 2, it can be seen that ethyl acetate lacks sufficient interfacial activity compared to 1-butyl-3-methylimidazole hexafluorophosphate, cannot effectively reduce the oil-water interfacial tension, and does not have a synergistic solubilization effect with lecithin, so it is difficult for the emulsified system to maintain long-term stability, which in turn causes the oil phase to stratify and precipitate.

[0088] The wall breaking efficiency test: The surface structure of the Ganoderma lucidum spores before and after the treatment with the extraction system of Example 2 and Comparative Example 1 was observed using a scanning electron microscope, the integrity and rupture of the spore wall were recorded, and the wall breaking rate was calculated as follows: the number of ruptured spores / the total number of spores × 100%. Figure 3 shown.

[0089] Depend on Figure 3 The results show that the spore wall breaking rate of the present invention is as high as 98.26%, which is extremely efficient. From the results of Comparative Example 1 and Example 2, it can be seen that although ultrasound and ionic liquid can still provide a certain wall breaking effect, the physical shearing and local micro-heating effects of nanoparticles are lacking, and the rupture of the spore wall only depends on ultrasound and chemical dissolution, lacking physical synergy, and the wall breaking efficiency is significantly affected.

[0090] Acid value determination: 1 mL of the Ganoderma lucidum spore oil sample prepared in Example 2 and Comparative Examples 1 to 4 was weighed and dissolved in 50 mL of ethanol-ether mixed solution, 3 drops of phenolphthalein indicator were added, and the mixture was titrated with 0.1 mol / L KOH solution. The consumed volume of KOH solution was recorded, and the acid value (mg KOH / mL) was calculated as follows: (KOH volume × KOH concentration × 56.1) / sample volume. Figure 4 shown.

[0091] Depend on Figure 4As a result, it can be seen that the ganoderma lucidum spore oil prepared by the extraction of the present invention through multiple synergistic effects has a low acid value, good oil quality, and the degradation of unsaturated fatty acids is effectively inhibited; from the results of comparative example 1 and example 2, it can be seen that due to the poor heat dispersion and oil release efficiency during the extraction process, the oil is exposed to a larger area and longer time in the system, which may increase the formation of free fatty acids, thereby leading to a higher acid value; from the results of comparative example 2 and example 2, it can be seen that the lack of ethoxylated soybean lecithin leads to a weakening of the emulsification and solubilization effects in the extraction system, the stability of the oil in the aqueous solution is reduced, and the oil is easy to agglomerate or precipitate, which may increase the oxidation and degradation of the oil, thereby leading to an increase in the acid value; from the results of comparative example 3 and example 2, it can be seen that the acid value is slightly affected; from the results of comparative example 4 and example 2, it can be seen that the oil extraction efficiency of ethyl acetate is much lower than that of ionic liquids, and its synergy with ethoxylated soybean lecithin is poor, the oil release rate is relatively slow, and the oil needs more time in the extraction system to enter the emulsified state, resulting in it being exposed to oxygen for a longer time during the whole process, thereby leading to an increase in the acid value caused by oxidation.

[0092] DPPH scavenging rate test: Take 2mL DPPH solution and 2mL sample solution to be tested (0.1mL of Ganoderma lucidum spore oil prepared in Examples 1 to 4 and Comparative Examples 1 to 4 is dissolved in 5mL of anhydrous ethanol), mix well, place in dark for 30min, and measure the absorbance at 517nm with anhydrous ethanol solution as reference. DPPH solution is a solution with a concentration of 0.1mM prepared with anhydrous ethanol as solvent. Calculate DPPH free radical scavenging rate (scavenging rate = [1-(absorbance of the mixture of sample solution to be tested and DPPH-absorbance of the sample solution without DPPH) / absorbance of the solution containing only DPPH] × 100%), repeat the test 3 times, take the average value, and the results are as follows: Figure 5 shown.

[0093] Depend on Figure 5As a result, the ganoderma lucidum spore oil prepared by the present invention has an excellent free radical scavenging effect; as can be seen from the results of comparative example 1 and example 2, the absence of modified alumina nanoparticles reduces the wall breaking efficiency, reduces the release of less oil and active ingredients, and further affects the free radical scavenging effect; as can be seen from the results of comparative example 2 and example 2, the absence of ethoxylated soybean lecithin affects the emulsification and dissolution of oil, resulting in reduced release and dissolution efficiency of active ingredients, and reduced content of antioxidant components in the extracted ganoderma lucidum spore oil; as can be seen from the results of comparative example 3 and example 2, the solubilization and emulsification effects of ordinary soybean lecithin are not as good as those of ethoxylated soybean lecithin, which affects the stability of oil and the dissolution efficiency of active ingredients during the extraction process, resulting in a slight decrease in antioxidant activity; as can be seen from the results of comparative example 4 and example 2, ethyl acetate, as a conventional organic solvent, has poor solubility and synergistic performance, low dissolution efficiency of active ingredients, and less active ingredients in the extracted oil, which further leads to poor free radical scavenging effect.

[0094] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An efficient extraction method for Ganoderma lucidum spore oil, characterized in that: The specific preparation steps are as follows: S1: Grinding the spore powder of Ganoderma lucidum through an 80-mesh sieve and drying it to obtain pretreated spore powder of Ganoderma lucidum; uniformly mixing the modified alumina nanoparticles and the pretreated spore powder of Ganoderma lucidum, and then adding a 5-10wt% ethoxylated soybean lecithin solution, stirring and mixing to obtain a mixed solution; S2: slowly adding the 1-butyl-3-methylimidazolium hexafluorophosphate solution to the mixed solution prepared in step S1, stirring at a speed of 500 rpm to mix evenly, and then stirring at a speed of 200-300 rpm, maintaining the temperature at 35° C., and continuously stirring for 20-30 minutes to obtain an extraction system; S3: placing the extraction system in step S2 in an ultrasonic reactor, and after the ultrasonic treatment is completed, cooling the extract to room temperature, centrifuging, filtering with a 0.22 μm filter membrane, washing with ethanol, and vacuum drying at 30° C. for 24 h to obtain Ganoderma lucidum spore oil.

2. The method for efficiently extracting ganoderma lucidum spore oil according to claim 1, characterized in that: The specific preparation steps of the modified aluminum oxide nanoparticles described in step S1 are as follows: A1: dissolving 3-aminopropyltriethoxysilane in anhydrous ethanol, stirring and dissolving uniformly to obtain a 3-aminopropyltriethoxysilane solution; suspending alumina nanoparticles in anhydrous ethanol, ultrasonically treating, centrifuging, washing with anhydrous ethanol, and calcining in an oven to obtain activated alumina nanoparticles; A2: slowly adding the activated alumina nanoparticles prepared in step A1 to the 3-aminopropyltriethoxysilane solution prepared in step A1, heating and stirring to react, centrifuging, and washing with ethanol to obtain pretreated alumina nanoparticles; A3: The pretreated alumina nanoparticles prepared in step A2 and succinic anhydride are added to ethyl acetate for mixing, reacting, washing with ethanol, centrifuging, and vacuum drying to constant weight to obtain modified alumina nanoparticles.

3. A highly efficient method for extracting Ganoderma lucidum spore oil according to claim 2, characterized in that: The ratio of the activated aluminum oxide nanoparticles to the 3-aminopropyltriethoxysilane solution used in step A2 is 1 g: 2-3 mL.

4. The efficient extraction method of Ganoderma lucidum spore oil according to claim 3, characterized in that: The ratio of the amount of the pretreated aluminum oxide nanoparticles, succinic anhydride and ethyl acetate used in step A3 is 1 g: 0.2-0.5 g: 20-30 mL.

5. The efficient extraction method of Ganoderma lucidum spore oil according to claim 4, characterized in that: The specific preparation steps of the ethoxylated soybean lecithin described in step S1 are as follows: B1: Add soybean lecithin to the ethanol solution, stir and mix evenly to obtain a soybean lecithin solution; add the soybean lecithin solution to a reaction kettle, add sodium hydroxide, seal and introduce nitrogen to maintain an inert atmosphere, then slowly introduce ethylene oxide gas, control the temperature to 100°C, the pressure to 0.2-0.5MPa, and react for 2-4h; B2: After the reaction in step B1 is completed, the mixture is cooled to room temperature, diluted hydrochloric acid is added to neutralize the sodium hydroxide, the reaction product is taken out, washed with deionized water and acetone, and vacuum dried to obtain ethoxylated soybean lecithin.

6. The method for efficiently extracting Ganoderma lucidum spore oil according to claim 5, characterized in that: The amount of the modified aluminum oxide nanoparticles in step S1 is 1.5-3% of the mass of the pretreated Ganoderma lucidum spore powder; the ratio of the amount of the ethoxylated soybean lecithin solution to the pretreated Ganoderma lucidum spore powder is 3mL:1g.

7. The method for efficiently extracting Ganoderma lucidum spore oil according to claim 6, characterized in that: The 1-butyl-3-methylimidazolium hexafluorophosphate solution in step S2 is prepared by mixing 1-butyl-3-methylimidazolium hexafluorophosphate and deionized water in a volume ratio of 1:4; the amount of 1-butyl-3-methylimidazolium hexafluorophosphate solution used is 15% of the volume of the mixed solution.

8. The method for efficiently extracting Ganoderma lucidum spore oil according to claim 7, characterized in that: The ultrasonic treatment parameters in step S3 are set to a frequency of 30-40 kHz, a power of 350-450 W, a system temperature of 32-37° C. maintained during the reaction, a treatment time of 30-50 min, and low-speed stirring at 200-300 rpm during the treatment.

Images