A ganoderma lucidum spore oil preparation method for reducing plasticizer residue

Abstract

The present application relates to a kind of ganoderma lucidum spore oil preparation methods for reducing plasticizer residual amount, the ganoderma lucidum spore oil preparation method includes the following steps: (1) raw material pre-adsorption: ganoderma lucidum spore powder raw material is mixed with modified chitosan microsphere and is treated with supercritical CO2 Extraction, after extraction treatment, remove modified chitosan microsphere, collect the plasticized ganoderma lucidum spore powder;(2) supercritical extraction: the plasticized ganoderma lucidum spore powder is treated with supercritical CO2 Extraction, collect the crude ganoderma lucidum spore oil;(3) adsorption refining: the crude ganoderma lucidum spore oil is mixed with modified activated carbon and modified diatomite, and is treated with adsorption, after filtration, obtain ganoderma lucidum spore oil.The preparation method can efficiently remove phthalate plasticizer in ganoderma lucidum spore oil with high selectivity, while retaining active ingredients such as ganoderma triterpenes, sterols, etc., process operation is simple and the obtained product is high in purity and stable in quality.

Description

Technical Field

[0001] This invention belongs to the field of Ganoderma lucidum spore oil preparation technology, and relates to a method for preparing Ganoderma lucidum spore oil with reduced plasticizer residue. Background Technology

[0002] Ganoderma lucidum spore oil is a fat-soluble active substance extracted from Ganoderma lucidum spore powder. It is rich in Ganoderma lucidum triterpenes, sterols, unsaturated fatty acids and other components. It has pharmacological activities such as enhancing immunity, inhibiting tumor cell growth and reducing the toxic side effects of radiotherapy and chemotherapy. However, during the harvesting, packaging, transportation and storage of Ganoderma lucidum spore powder, plastic products such as plastic films, plastic bags and plastic collectors are often used. Phthalate plasticizers in these products can easily migrate and contaminate the Ganoderma lucidum spore powder.

[0003] In existing technologies, the removal of plasticizers from Ganoderma lucidum spore oil mainly relies on direct mixed adsorption with a single adsorbent or simple supercritical CO2 extraction separation. For example, CN121852130A discloses a purification method for plasticizer-free Ganoderma lucidum spore oil, including the following steps: raw material pretreatment, supercritical CO2 fluid extraction, desorption separation, and post-treatment; the supercritical fluid extraction step uses supercritical carbon dioxide fluid with adjustable polarity, combined with the use of an entrainer. Through the selective dissolution characteristics of the fluid on plasticizer molecules, plasticizers are preferentially removed during the extraction process. The desorption separation step also includes adsorption treatment, in which an adsorbent is added to the separation vessel to further remove trace amounts of plasticizers; the adsorbent is at least one of activated carbon, silica gel, or molecular sieve.

[0004] However, plasticizers and active ingredients such as Ganoderma lucidum triterpenes and sterols have similar solubility behaviors and polar distributions in supercritical CO2. It is difficult to achieve selective separation by simply relying on pressure and temperature control, which can easily lead to the loss of active ingredients. At the same time, traditional adsorbents such as activated carbon are prone to local agglomeration and adsorption saturation in the oil phase. After saturation, the adsorbed plasticizers will be re-desorbed and released during subsequent processing or storage, causing secondary pollution. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention aims to provide a method for preparing Ganoderma lucidum spore oil with reduced plasticizer residue.

[0006] To achieve this objective, the present invention adopts the following technical solution:

[0007] This invention provides a method for preparing Ganoderma lucidum spore oil with reduced plasticizer residue, the method comprising the following steps:

[0008] (1) Pre-adsorption of raw materials: Ganoderma lucidum spore powder raw material is mixed with modified chitosan microspheres and subjected to supercritical CO2 extraction. After extraction, the modified chitosan microspheres are separated and removed, and the deplasticized Ganoderma lucidum spore powder is collected.

[0009] (2) Supercritical extraction: The deplasticized Ganoderma lucidum spore powder was subjected to supercritical CO2 extraction to collect crude Ganoderma lucidum spore oil;

[0010] (3) Adsorption purification: Ganoderma lucidum spore oil crude product is mixed with modified activated carbon and modified diatomaceous earth, and adsorption treatment is carried out. After filtration, Ganoderma lucidum spore oil is obtained.

[0011] To address the technical problem of phthalate plasticizers easily migrating and contaminating Ganoderma lucidum spore powder, resulting in high plasticizer residues in the prepared Ganoderma lucidum spore oil product, this invention creatively employs a method of sequentially pre-adsorbing modified chitosan microspheres onto Ganoderma lucidum spore powder raw materials, supercritical CO2 extraction, and then using a composite adsorbent of modified activated carbon and modified diatomaceous earth for synergistic adsorption treatment to prepare Ganoderma lucidum spore oil. This preparation method can efficiently and selectively remove phthalate plasticizers from Ganoderma lucidum spore oil while fully retaining active ingredients such as Ganoderma lucidum triterpenes and sterols. The process is simple to operate, and the obtained product has high purity and stable quality.

[0012] Preferably, the mass ratio of the modified chitosan microspheres to the Ganoderma lucidum spore powder raw material in step (1) is 100:(1-15), for example, 100:1, 100:3, 100:5, 100:7, 100:8, 100:10, 100:12, 100:13, 100:15, etc.

[0013] Preferably, the supercritical CO2 extraction process in step (1) is carried out under the following conditions: extraction pressure of 10-20 MPa (e.g., 10 MPa, 12 MPa, 14 MPa, 16 MPa, 18 MPa, 20 MPa, etc.), extraction temperature of 35-45℃ (e.g., 35℃, 37℃, 38℃, 40℃, 45℃, etc.), and CO2 flow rate of 15-25 L / h (e.g., 15 L / h, 17 L / h, 18 L / h, 20 L / h, 25 L / h, etc.) for 1-4 h (e.g., 1 h, 2 h, 3 h, 4 h, etc.).

[0014] Preferably, the separation in step (1) is performed using a 10-20 mesh vibrating screen.

[0015] Preferably, the modified chitosan microspheres in step (1) have a particle size of 0.5-1.5 mm, such as 0.5 mm, 0.8 mm, 1.0 mm, 1.2 mm, 1.5 mm, etc.

[0016] Preferably, the method for preparing the modified chitosan microspheres includes:

[0017] Chitosan was treated with nano-silica sol, then mixed and emulsified with β-cyclodextrin and liquid paraffin containing Span-80. A crosslinking agent was then added to carry out the reaction. After the reaction was completed, the microspheres were collected by centrifugation, washed, dried, and sieved to obtain the modified chitosan microspheres.

[0018] Traditional chitosan adsorbents are easily broken and powdered during supercritical high-pressure dynamic extraction, making them difficult to separate from spore powder and prone to residues causing subsequent pollution. Furthermore, they lack specific recognition of plasticizers and exhibit poor adsorption selectivity. Therefore, this invention utilizes nano-silica intercalation, the introduction of β-cyclodextrin inclusion sites, and glutaraldehyde cross-linking to enable microspheres to possess high strength, millimeter-scale sieveable particle size, and selective recognition capability of cyclodextrin cavities. In the supercritical system, these microspheres preferentially adsorb plasticizers rather than Ganoderma triterpenes, significantly reducing the initial plasticizer concentration in the oil phase while avoiding excessive loss of active ingredients. They can also be easily recovered using a vibrating screen without the risk of residual pollution.

[0019] In this invention, modified chitosan microspheres enhance mechanical strength through nano-silica intercalation, provide selective inclusion sites through β-cyclodextrin, and form a three-dimensional covalent network through glutaraldehyde cross-linking. This maintains a complete spherical structure during supercritical CO2 high-pressure dynamic extraction, avoiding the defects of traditional chitosan adsorbents such as easy breakage and difficulty in separation. Their millimeter-sized particles can be easily recovered using a vibrating screen, preventing residual powder from contaminating subsequent extractions. The cyclodextrin cavities on the microsphere surface have a specific recognition ability for the benzene ring of phthalate plasticizers, achieving preferential adsorption in a supercritical system where plasticizers and Ganoderma lucidum triterpenes are co-soluble. This solves the problem of poor selectivity of conventional adsorbents. In synergy with subsequently modified activated carbon and modified diatomaceous earth, the initial concentration of plasticizers in the oil phase is significantly reduced, alleviating the adsorption load in the later stages. This achieves deep purification with low adsorbent dosage while avoiding excessive loss of active ingredients.

[0020] Preferably, the chitosan is treated with nano-silica sol as follows: chitosan is dissolved in an aqueous acetic acid solution with a volume fraction of 1.0-2.0% (e.g., 1.0%, 1.5%, 2.0%) at a mass ratio of 1:(50-100) (e.g., 1:50, 1:60, 1:70, 1:80, 1:90, 1:100, etc.). The chitosan acetic acid solution is then added to the pre-ultrasonically dispersed nano-silica sol, and the mixture is stirred at 200-400 rpm (e.g., 200 rpm, 250 rpm, 300 rpm, 350 rpm, 400 rpm, etc.) at 200-30℃ (e.g., 20℃, 22℃, 25℃, 28℃, 30℃, etc.) for 30-60 min (e.g., 30 min, 40 min, 50 min, 60 min, etc.).

[0021] Preferably, the mass ratio of the nano-silica to chitosan is 1:(5-10), for example, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0022] Preferably, the mass ratio of nano-silica to deionized water in the nano-silica sol is 1:(5-10), for example, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0023] Preferably, the mixed emulsification method is as follows: chitosan treated with nano-silica sol is mixed with β-cyclodextrin and stirred at 30-40℃ (e.g., 30℃, 32℃, 35℃, 38℃, 40℃, etc.) for 1-2 h (e.g., 1 h, 1.2 h, 1.5 h, 1.8 h, 2 h, etc.) to form a composite aqueous phase, which is then mixed with liquid paraffin containing Span-80 as the oil phase, and emulsified at 8000-12000 rpm (e.g., 8000 rpm, 9000 rpm, 10000 rpm, 11000 rpm, 12000 rpm, etc.) for 10-20 min (e.g., 10 min, 15 min, 20 min, etc.) to form a W / O type emulsion.

[0024] Preferably, the mass ratio of β-cyclodextrin to chitosan is 1:(2-5), for example, 1:2, 1:3, 1:5, etc.

[0025] Preferably, the volume ratio of Span-80 to liquid paraffin is 1:(20-50), for example, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50, etc.

[0026] Preferably, the volume ratio of the aqueous phase to the oil phase is 1:(3-5), for example, 1:3, 1:4, 1:5, etc.

[0027] Preferably, the crosslinking agent is an aqueous solution of glutaraldehyde with a mass fraction of 23-27%.

[0028] Preferably, the mass ratio of glutaraldehyde to chitosan is (0.3-0.9):1, for example, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, etc.

[0029] Preferably, the reaction is carried out at 40-50°C (e.g., 40°C, 45°C, 50°C, etc.) for 3-5 h (e.g., 3 h, 3.5 h, 4 h, 4.5 h, 5 h, etc.).

[0030] Preferably, the supercritical CO2 extraction treatment in step (2) is carried out under the following conditions: static extraction for 20-70 min (e.g., 20 min, 30 min, 33 MPa, 35 MPa, etc.), extraction temperature for 40-50℃ (e.g., 40℃, 42℃, 45℃, 48℃, 50℃, etc.), and CO2 flow rate for 20-30 L / h (e.g., 20 L / h, 23 L / h, 25 L / h, 28 L / h, 30 L / h, etc.), followed by dynamic extraction for 1-4 h (e.g., 1 h, 2 h, 3 h, 4 h, etc.).

[0031] Preferably, the mass ratio of modified activated carbon to modified diatomaceous earth in step (3) is (2-5):1, for example, 2:1, 3:1, 4:1, 5:1, etc.

[0032] Preferably, the total amount of modified activated carbon and modified diatomaceous earth added is 1-5% of the crude Ganoderma lucidum spore oil, such as 1%, 2%, 3%, 4%, 5%, etc.

[0033] Preferably, the adsorption treatment in step (3) is carried out at 30-40℃ (e.g., 30℃, 35℃, 40℃, etc.) for 1-3 h (e.g., 1 h, 1.5 h, 2 h, 2.5 h, 3 h, etc.).

[0034] Preferably, the filtration in step (3) is performed using a plate and frame filter press.

[0035] Preferably, the method for preparing the modified activated carbon in step (3) includes:

[0036] Activated carbon was mixed with nitric acid solution for oxidation, washed, and dried to obtain oxidized activated carbon; the oxidized activated carbon was dispersed in an aqueous ethanol solution and mixed with a silane coupling agent for silanization, filtered, washed, and dried to obtain aminated activated carbon; the aminated activated carbon was dispersed in deionized water and mixed with citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride for amidation, filtered, washed, and dried to obtain the modified activated carbon.

[0037] Traditional activated carbon relies solely on physical pores for adsorption, which is easily saturated. Adsorbed plasticizers can desorb during storage or subsequent processing, causing secondary pollution. Furthermore, the pores are easily blocked by large molecules. Therefore, this invention employs a three-step chemical modification process—oxidation, silanization, and amidation—to graft small-molecule citric acid onto the surface, constructing a high-density carboxyl / amino polar functional group. This group forms a strong chemical affinity with the plasticizer ester groups through hydrogen bonds and ion-dipole interactions, achieving firm fixation and preventing desorption. The grafted molecules are small in size, do not clog micropores, and maintain a high specific surface area, thereby deeply capturing trace amounts of plasticizers and preventing secondary release.

[0038] In this invention, modified activated carbon adsorbents are oxidized with nitric acid to introduce carboxyl and phenolic hydroxyl groups, silanized and grafted with amino groups, and citric acid is amidated and grafted with small molecule carboxylic acids under EDC catalysis, constructing a high-density polar functional group surface. Unlike traditional activated carbon, which relies solely on physical pores for adsorption and is prone to saturation and desorption of plasticizers, this modified activated carbon forms a strong chemical affinity with the ester groups of plasticizers through hydrogen bonds, ions, and dipole interactions. Moreover, the grafted molecules are small in size and do not clog micropores, maintaining a high specific surface area. During the oil phase refining stage, this modified activated carbon can capture trace amounts of plasticizers that were not completely removed in the early stages and firmly fix them, avoiding secondary release during storage or subsequent processing. When used in combination with modified diatomaceous earth, the surface polarities of the two are complementary. The activated carbon deeply adsorbs, while the diatomaceous earth provides a filtration channel and assists in adsorption, synergistically improving filtration efficiency and oil clarity.

[0039] Preferably, the activated carbon has a particle size of 0.5-1.0 mm.

[0040] Preferably, the mass fraction of the nitric acid solution is 10-15% (e.g., 10%, 11%, 12%, 13%, 14%, 15%, etc.), and the mass ratio of the activated carbon to the nitric acid solution is 1:(5-10), e.g., 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0041] Preferably, the oxidation reaction is carried out at 70-80℃ (e.g., 70℃, 72℃, 75℃, 78℃, 80℃, etc.) for 2-3 hours (e.g., 2 hours, 2.2 hours, 2.5 hours, 2.8 hours, 3 hours, etc.).

[0042] Preferably, the mass ratio of the oxidizing activated carbon to the ethanol aqueous solution is 1:(8-15) (e.g., 1:8, 1:9, 1:10, 1:12, 1:14, 1:15, etc.), and the volume fraction of the ethanol aqueous solution is 85-95% (e.g., 85%, 90%, 95%, etc.).

[0043] Preferably, the silane coupling agent is KH-550, and the mass ratio of KH-550 to oxidizing activated carbon is (0.1-0.3):1, for example, 0.1:1, 0.15:1, 0.2:1, 0.25:1, 0.3:1, etc.

[0044] Preferably, the silanization reaction is carried out at 60-70°C (e.g., 60°C, 62°C, 65°C, 68°C, 70°C, etc.) for 4-6 h (e.g., 4 h, 4.5 h, 5 h, 5.5 h, 6 h, etc.).

[0045] Preferably, the amidation reaction is carried out at pH 5.0-6.0 (e.g., pH 5.0, pH 5.2, pH 5.4, pH 5.8, pH 5.0, etc.) and 25-35°C (e.g., 25°C, 28°C, 30°C, 32°C, 35°C, etc.) for 4-8 h (e.g., 4 h, 5 h, 6 h, 7 h, 8 h, etc.).

[0046] Preferably, the method for preparing the modified diatomite in step (3) includes:

[0047] Diatomaceous earth is mixed with an acid solution for acid treatment, and then the acid-treated diatomaceous earth is mixed with an alkaline solution for alkali dissolution reaction. After filtration, washing, and drying, activated diatomaceous earth is obtained. The activated diatomaceous earth is dispersed in an ethanol aqueous solution and mixed with a silane coupling agent for silanization reaction. After filtration, washing, and drying, aminated diatomaceous earth is obtained. The aminated diatomaceous earth is dispersed in deionized water and mixed with tannic acid for reaction. After filtration, washing, and drying, the modified diatomaceous earth is obtained.

[0048] Traditional diatomaceous earth is only used as a physical filter aid and has no ability to remove plasticizers. Furthermore, it easily causes activated carbon particles to agglomerate during filtration, reducing filtration efficiency. This invention addresses this by using acid-base activation to expose silanol groups, grafting amino groups with silane coupling agents, and oxidative covalent grafting of tannic acid to form a polyphenol functional layer. Utilizing the multiple hydrogen bonds and π-π stacking effects of the pyrogallol structure on plasticizers, diatomaceous earth gains both adsorption and filtration aid functions. Its rigid porous framework prevents activated carbon agglomeration, forming a uniform composite filter cake layer, accelerating plate and frame filtration, shortening the production cycle, and reducing energy consumption. Moreover, the covalently fixed tannic acid layer is heat-resistant, oil-resistant, and does not detach or contaminate oil products.

[0049] In this invention, the modified diatomaceous earth filter aid is activated by acid and alkali to expose silanol groups, introduces amino groups with silane coupling agents, and forms a polyphenol functional layer through tannic acid oxidative grafting in alkaline air. The pyrogallol structure utilizes multiple hydrogen bonds and π-π stacking effects on the ester groups of plasticizers to achieve auxiliary adsorption, while maintaining the original micron-scale pore structure of diatomaceous earth, thus improving filtration throughput. Unlike traditional filter aids that only perform physical filtration and have no ability to remove plasticizers, this modified diatomaceous earth has both adsorption and filtration functions, reducing the amount of additional adsorbent required. Its surface covalently fixed tannic acid layer is heat-resistant and oil-soluble, and will not detach and contaminate oil products. When used in conjunction with modified activated carbon, the porous rigid framework of diatomaceous earth can prevent activated carbon particles from agglomerating, forming a uniform composite filter cake layer, accelerating the plate and frame filtration speed, shortening the production cycle, and reducing energy consumption.

[0050] Preferably, the particle size of the diatomaceous earth is 10-50 μm.

[0051] Preferably, the acid solution is a hydrochloric acid solution of 0.5-1.5 mol / L (e.g., 0.5 mol / L, 0.8 mol / L, 1.0 mol / L, 1.2 mol / L, 1.5 mol / L, etc.), and the mass ratio of the diatomaceous earth to the acid solution is 1:(5-10), for example, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0052] Preferably, the acid treatment is carried out at 80-90℃ (e.g., 80℃, 82℃, 85℃, 88℃, 90℃, etc.) with stirring for 1-2 h (e.g., 1 h, 1.2 h, 1.5 h, 1.8 h, 2 h, etc.).

[0053] Preferably, the alkaline solution is a sodium hydroxide solution of 0.5-1.5 mol / L (e.g., 0.5 mol / L, 0.8 mol / L, 1.0 mol / L, 1.2 mol / L, 1.5 mol / L, etc.), and the mass ratio of the acid-treated diatomaceous earth to the alkaline solution is 1:(5-10), for example, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0054] Preferably, the alkaline dissolution reaction is carried out at 80-90℃ (e.g., 80℃, 82℃, 85℃, 88℃, 90℃, etc.) with stirring for 1-2 hours (e.g., 1 hour, 1.2 hours, 1.5 hours, 1.8 hours, 2 hours, etc.).

[0055] Preferably, the volume fraction of the ethanol-water solution is 85-95% (e.g., 85%, 90%, 95%, etc.), and the mass ratio of the activated diatomaceous earth to the ethanol-water solution is 1:(8-15) (e.g., 1:8, 1:9, 1:10, 1:12, 1:14, 1:15, etc.).

[0056] Preferably, the silane coupling agent is KH-550, and the mass ratio of KH-550 to activated diatomaceous earth is (0.1-0.3):1, for example, 0.1:1, 0.15:1, 0.2:1, 0.25:1, 0.3:1, etc.

[0057] Preferably, the silanization reaction is carried out at 60-70°C (e.g., 60°C, 62°C, 65°C, 68°C, 70°C, etc.) for 4-6 h (e.g., 4 h, 4.5 h, 5 h, 5.5 h, 6 h, etc.).

[0058] Preferably, the mass ratio of the aminated diatomaceous earth to deionized water is 1:(10-20), such as 1:10, 1:12, 1:14, 1:16, 1:18, 1:20, etc.

[0059] Preferably, the reaction with tannic acid is carried out at pH=8.0-9.0 (e.g., pH=8.0, pH=8.2, pH=8.4, pH=8.8, pH=9.0, etc.) and 50-60℃ (e.g., 50℃, 52℃, 55℃, 58℃, 60℃, etc.) for 8-12 h (e.g., 8 h, 9 h, 10 h, 11 h, 12 h, etc.), and the mass ratio of tannic acid to aminated diatomaceous earth is (0.3-0.5):1 (e.g., 0.3:1, 0.35:1, 0.4:1, 0.45:1, 0.5:1, etc.).

[0060] All other unlisted point values ​​within the above numerical ranges can be selected, and all are within the protection scope of this invention, and will not be elaborated here.

[0061] Compared with the prior art, the present invention has the following beneficial effects:

[0062] To address the technical problem of phthalate plasticizers easily migrating and contaminating Ganoderma lucidum spore powder, resulting in high plasticizer residues in the prepared Ganoderma lucidum spore oil product, this invention creatively employs a method of sequentially pre-adsorbing modified chitosan microspheres onto Ganoderma lucidum spore powder raw materials, supercritical CO2 extraction, and then using a composite adsorbent of modified activated carbon and modified diatomaceous earth for synergistic adsorption treatment to prepare Ganoderma lucidum spore oil. This preparation method can efficiently and selectively remove phthalate plasticizers from Ganoderma lucidum spore oil while fully retaining active ingredients such as Ganoderma lucidum triterpenes and sterols. The process is simple to operate, and the obtained product has high purity and stable quality. Detailed Implementation

[0063] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0064] Preparation Example 1-1

[0065] This preparation example provides a modified chitosan microsphere, which is prepared by the following method:

[0066] (1) Chitosan was dissolved in an aqueous solution of 1.0% acetic acid at a ratio of 1:50 g / g and stirred at 300 rpm for 1 h at 20 °C to obtain a chitosan acetic acid solution.

[0067] (2) Add the chitosan acetic acid solution to the pre-ultrasonic dispersed nano silica sol, stir at 200 rpm for 30 min at 20 °C, and embed nano SiO2 into the chitosan molecular chains through hydrogen bonding and physical embedding; the mass ratio of nano silica to chitosan is 1:5; the mass ratio of nano silica to deionized water in the nano silica sol is 1:5.

[0068] (3) Add β-cyclodextrin and continue stirring at 30°C for 1 hour to form a pre-composite system as the aqueous phase through intermolecular hydrogen bonds; liquid paraffin containing Span-80 is used as the oil phase. Mix the aqueous phase and oil phase and emulsify at 8000 rpm for 10 minutes to form a W / O type emulsion; the mass ratio of β-cyclodextrin to chitosan is 1:2; the volume ratio of Span-80 to liquid paraffin is 1:20; and the volume ratio of the aqueous phase to the oil phase is 1:3.

[0069] (4) Then add 23% glutaraldehyde aqueous solution, with a mass ratio of glutaraldehyde to chitosan of 0.3:1. Stir at 200 rpm for 3 h at 40 °C to form a Schiff base reaction (glutaraldehyde aldehyde group reacts with chitosan amino group to form an imine bond) and a hemiacetal reaction (glutaraldehyde aldehyde group reacts with β-cyclodextrin hydroxyl group) to form a three-dimensional covalent cross-linked network.

[0070] (5) After the reaction was completed, the microspheres were collected by centrifugation at 3000 rpm for 5 min. They were washed twice with petroleum ether and anhydrous ethanol, and three times with deionized water. Finally, they were dried at 40℃ and vacuum degree -0.08MPa for 12 h, passed through a 10-mesh sieve, and the microspheres with a particle size of 0.5 mm were collected to obtain modified chitosan microspheres.

[0071] Preparation Examples 1-2

[0072] This preparation example provides a modified chitosan microsphere, which is prepared by the following method:

[0073] (1) Chitosan was dissolved in a 1.5% acetic acid aqueous solution at a material-to-liquid ratio of 1:75 g / g and stirred at 400 rpm for 1.5 h at 25 °C to obtain a chitosan acetic acid solution;

[0074] (2) Add the chitosan acetic acid solution to the pre-ultrasonic dispersed nano silica sol, stir at 300 rpm for 45 min at 25 °C, and embed nano SiO2 into the chitosan molecular chains through hydrogen bonding and physical embedding; the mass ratio of nano silica to chitosan is 1:7.5; the mass ratio of nano silica to deionized water in the nano silica sol is 1:7.5;

[0075] (3) Add β-cyclodextrin and continue stirring at 35°C for 1.5 h to form a pre-composite system as the aqueous phase through intermolecular hydrogen bonds; liquid paraffin containing Span-80 is used as the oil phase. Mix the aqueous phase and oil phase and emulsify at 10,000 rpm for 15 min to form a W / O type emulsion; the mass ratio of β-cyclodextrin to chitosan is 1:3.5; the volume ratio of Span-80 to liquid paraffin is 1:35; and the volume ratio of the aqueous phase to the oil phase is 1:4.

[0076] (4) Then add 25% glutaraldehyde aqueous solution, with a mass ratio of glutaraldehyde to chitosan of 0.6:1. Stir at 250 rpm for 4 h at 45 °C to form a Schiff base reaction (glutaraldehyde aldehyde group reacts with chitosan amino group to form an imine bond) and a hemiacetal reaction (glutaraldehyde aldehyde group reacts with β-cyclodextrin hydroxyl group) to form a three-dimensional covalent cross-linked network.

[0077] (5) After the reaction was completed, the microspheres were collected by centrifugation at 3500 rpm for 7 min. They were washed twice with petroleum ether and anhydrous ethanol, and three times with deionized water. Finally, they were dried at 45℃ and vacuum degree -0.09MPa for 18 h, passed through a 15-mesh sieve, and microspheres with a particle size of 1 mm were collected to obtain modified chitosan microspheres.

[0078] Preparation Examples 1-3

[0079] This preparation example provides a modified chitosan microsphere, which is prepared by the following method:

[0080] (1) Chitosan was dissolved in an aqueous solution of 2.0% acetic acid at a ratio of 1:100 g / g and stirred at 500 rpm for 2 h at 30 °C to obtain a chitosan acetic acid solution.

[0081] (2) Add the chitosan acetic acid solution to the pre-ultrasonic dispersed nano silica sol, stir at 400 rpm for 60 min at 30 °C, and embed nano SiO2 into the chitosan molecular chains through hydrogen bonding and physical embedding; the mass ratio of nano silica to chitosan is 1:10; the mass ratio of nano silica to deionized water in the nano silica sol is 1:10.

[0082] (3) Add β-cyclodextrin and continue stirring at 40°C for 2 hours to form a pre-composite system as the aqueous phase through intermolecular hydrogen bonds; liquid paraffin containing Span-80 is used as the oil phase. Mix the aqueous phase and oil phase and emulsify at 12000 rpm for 20 minutes to form a W / O type emulsion; the mass ratio of β-cyclodextrin to chitosan is 1:5; the volume ratio of Span-80 to liquid paraffin is 1:50; and the volume ratio of the aqueous phase to the oil phase is 1:5.

[0083] (4) Then add 27% glutaraldehyde aqueous solution, with a mass ratio of glutaraldehyde to chitosan of 0.9:1. Stir at 300 rpm for 5 h at 50 °C to form a Schiff base reaction (glutaraldehyde aldehyde group reacts with chitosan amino group to form an imine bond) and a hemiacetal reaction (glutaraldehyde aldehyde group reacts with β-cyclodextrin hydroxyl group) to form a three-dimensional covalent cross-linked network.

[0084] (5) After the reaction was completed, the microspheres were collected by centrifugation at 4000 rpm for 10 min. They were washed twice with petroleum ether and anhydrous ethanol, and three times with deionized water. Finally, they were dried at 50℃ and vacuum degree -0.1MPa for 24 h, passed through a 20-mesh sieve, and the microspheres with a particle size of 1.5 mm were collected to obtain modified chitosan microspheres.

[0085] Preparation Example 2-1

[0086] This preparation example provides a modified activated carbon adsorbent, the preparation method of which is as follows:

[0087] (1) Add activated carbon (particle size 0.5 mm) to a 10% nitric acid solution at a material-to-liquid ratio of 1:5 g / g. In a constant temperature water bath at 70℃, mechanically stir at 200 rpm for 2 h for oxidation reaction (nitric acid oxidizes carbon atoms on the surface of activated carbon to carboxyl-COOH and phenolic hydroxyl-OH). After the reaction is completed, wash with deionized water until the pH of the filtrate is 6.5. Dry at 100℃ for 4 h to obtain oxidized activated carbon.

[0088] (2) Disperse the oxidized activated carbon in a 90% volume fraction ethanol aqueous solution (the ratio of oxidized activated carbon to ethanol aqueous solution is 1:8 g / g), add KH-550 (the mass ratio of KH-550 to oxidized activated carbon is 0.1:1), and carry out a silanization reaction at 60℃ for 4h (the ethoxy group of KH-550 dehydrates and condenses with the hydroxyl group on the surface of activated carbon to form a Si-OC covalent bond, and at the same time introduces amino-NH2). After the reaction is completed, filter, wash 3 times with anhydrous ethanol, and dry in a vacuum drying oven at 50℃ for 4h to obtain aminated activated carbon.

[0089] (3) Disperse the aminated activated carbon in deionized water (the ratio of aminated activated carbon to deionized water is 1:5 g / g), and add an aqueous solution containing citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride (EDC∙HCl) (where the concentration of citric acid is 0.3 mol / L, and the molar ratio of EDC∙HCl to citric acid is 0.5:1) (the ratio of aminated activated carbon to the aqueous solution containing citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride is 1:10). The modified activated carbon adsorbent was prepared by adjusting the pH to 5.0 with 0.1 mol / L hydrochloric acid and stirring at 150 rpm for 4 h at 25 °C. The reaction was carried out by amidation (EDC∙HCl activated the carboxyl group of citric acid, which formed an amide bond -CO-NH- with the amino group on the surface of activated carbon, thus grafting the small citric acid molecules onto the surface of activated carbon). After the reaction was completed, the mixture was filtered, washed three times with deionized water and twice with anhydrous ethanol, and dried at 50 °C for 8 h.

[0090] Preparation Example 2-2

[0091] This preparation example provides a modified activated carbon adsorbent, the preparation method of which is as follows:

[0092] (1) Activated carbon (particle size 0.7 mm) was added to a 12.5% ​​nitric acid solution at a material-to-liquid ratio of 1:7.5 g / g. The solution was then subjected to an oxidation reaction at 250 rpm in a constant temperature water bath at 75 °C for 2.5 h (nitric acid oxidizes carbon atoms on the surface of activated carbon to carboxyl-COOH and phenolic hydroxyl-OH). After the reaction was completed, the solution was washed with deionized water until the pH of the filtrate was 6.7. The solution was then dried at 105 °C for 5 h to obtain oxidized activated carbon.

[0093] (2) Disperse the oxidized activated carbon in a 90% volume fraction ethanol aqueous solution (the ratio of oxidized activated carbon to ethanol aqueous solution is 1:11.5 g / g), add KH-550 (the mass ratio of KH-550 to oxidized activated carbon is 0.2:1), and carry out a silanization reaction at 65℃ for 5h (the ethoxy group of KH-550 dehydrates and condenses with the hydroxyl group on the surface of activated carbon to form Si-OC covalent bond, and at the same time introduces amino-NH2). After the reaction is completed, filter, wash 3 times with anhydrous ethanol, and dry in a vacuum drying oven at 55℃ for 6h to obtain aminated activated carbon.

[0094] (3) Disperse the aminated activated carbon in deionized water (the ratio of aminated activated carbon to deionized water is 1:7.5 g / g), and add an aqueous solution containing citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride (EDC∙HCl) (where the concentration of citric acid is 0.4 mol / L, and the molar ratio of EDC∙HCl to citric acid is 0.7:1) (the ratio of aminated activated carbon to the aqueous solution containing citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride is 1:15). The modified activated carbon adsorbent was prepared by adjusting the pH to 5.5 with 0.1 mol / L hydrochloric acid and stirring at 200 rpm for 6 h at 30 °C. The reaction was carried out by amidation (EDC∙HCl activated the carboxyl group of citric acid, which formed an amide bond -CO-NH- with the amino group on the surface of activated carbon, thus grafting the small citric acid molecules onto the surface of activated carbon). After the reaction was completed, the mixture was filtered, washed three times with deionized water and twice with anhydrous ethanol, and dried at 55 °C for 10 h.

[0095] Preparation Examples 2-3

[0096] This preparation example provides a modified activated carbon adsorbent, the preparation method of which is as follows:

[0097] (1) Add activated carbon (particle size 1.0 mm) to a 15% nitric acid solution at a material-to-liquid ratio of 1:10 g / g. In a constant temperature water bath at 80℃, mechanically stir at 300 rpm for 3 h for oxidation reaction (nitric acid oxidizes carbon atoms on the surface of activated carbon to carboxyl-COOH and phenolic hydroxyl-OH). After the reaction is completed, wash with deionized water until the pH of the filtrate is 7.0. Dry at 110℃ for 6 h to obtain oxidized activated carbon.

[0098] (2) Disperse the oxidized activated carbon in a 90% volume fraction ethanol aqueous solution (the ratio of oxidized activated carbon to ethanol aqueous solution is 1:15 g / g), add KH-550 (the mass ratio of KH-550 to oxidized activated carbon is 0.3:1), and carry out a silanization reaction at 70℃ for 6h (the ethoxy group of KH-550 dehydrates and condenses with the hydroxyl group on the surface of activated carbon to form a Si-OC covalent bond, and at the same time introduces amino-NH2). After the reaction is completed, filter, wash 3 times with anhydrous ethanol, and dry in a vacuum drying oven at 60℃ for 8h to obtain aminated activated carbon.

[0099] (3) Disperse the aminated activated carbon in deionized water (the ratio of aminated activated carbon to deionized water is 1:10 g / g), and add an aqueous solution containing citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride (EDC∙HCl) (where the concentration of citric acid is 0.6 mol / L, and the molar ratio of EDC∙HCl to citric acid is 1:1) (the ratio of aminated activated carbon to the aqueous solution containing citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride is 1:20). The modified activated carbon adsorbent was prepared by adjusting the pH to 6.0 with 0.1 mol / L hydrochloric acid and stirring at 250 rpm for 8 h at 35 °C. The reaction was carried out by amidation (EDC∙HCl activated the carboxyl group of citric acid, which formed an amide bond -CO-NH- with the amino group on the surface of activated carbon, thus grafting the small citric acid molecules onto the surface of activated carbon). After the reaction was completed, the mixture was filtered, washed three times with deionized water and twice with anhydrous ethanol, and dried at 60 °C for 12 h.

[0100] Preparation Example 3-1

[0101] This preparation example provides a modified diatomaceous earth filter aid, the preparation method of which is as follows:

[0102] (1) Add diatomaceous earth (particle size 10 μm) to 1 mol / L hydrochloric acid solution at a material-to-liquid ratio of 1:5. Stir at 200 rpm for 1 h in a constant temperature water bath at 80℃ to undergo acid dissolution reaction (dissolving impurities such as carbonates and metal oxides). After the reaction is completed, filter and wash with deionized water until the pH of the filtrate is 6.5. Then add the acid-treated diatomaceous earth to 1 mol / L sodium hydroxide solution at a material-to-liquid ratio of 1:5 (g / g). Stir at 80℃ for 1 h to undergo alkali dissolution reaction (dissolving amorphous silica and some impurities, exposing more silanol groups -Si-OH). After the reaction is completed, filter and wash with deionized water until the pH of the filtrate is 6.5. Dry at 100℃ for 4 h to obtain activated diatomaceous earth.

[0103] (2) Activated diatomaceous earth was dispersed in a 90% ethanol aqueous solution at a material-to-liquid ratio of 1:8 g / g, KH-550 was added, and a silanization reaction was carried out at 60℃ for 4h (the mass ratio of KH-550 to activated diatomaceous earth was 0.1:1; the ethoxy group of KH-550 dehydrated and condensed with the silanol group on the surface of diatomaceous earth to form Si-O-Si covalent bonds, and amino-NH2 was introduced at the same time). After the reaction was completed, the mixture was filtered, washed three times with anhydrous ethanol, and dried at 50℃ for 4h to obtain aminated diatomaceous earth.

[0104] (3) Disperse aminated diatomaceous earth in deionized water at a material-to-liquid ratio of 1:10 g / g, add tannic acid aqueous solution (tannic acid to aminated diatomaceous earth mass ratio of 0.3:1), adjust pH to 8.0 with sodium bicarbonate, stir at 150 rpm at 50℃, and simultaneously introduce air (flow rate 0.2 L / min) for 8 h (tannic acid self-oxidizes in weakly alkaline air to generate quinone intermediate, which undergoes Michael addition reaction and Schiff base reaction with amino groups on the surface of diatomaceous earth, and covalently grafts tannic acid onto the surface of diatomaceous earth). After the reaction is completed, filter, wash 3 times with deionized water and 2 times with anhydrous ethanol, freeze dry at -20℃ and vacuum degree 0.1 mbar for 24 h (avoid thermal oxidation and deactivation of phenolic hydroxyl groups) to obtain modified diatomaceous earth filter aid.

[0105] Preparation Example 3-2

[0106] This preparation example provides a modified diatomaceous earth filter aid, the preparation method of which is as follows:

[0107] (1) Diatomaceous earth (particle size 30 μm) was added to 1 mol / L hydrochloric acid solution at a material-to-liquid ratio of 1:7.5. The solution was stirred at 250 rpm for 1.5 h in a constant temperature water bath at 85 °C to undergo an acid dissolution reaction (dissolving impurities such as carbonates and metal oxides). After the reaction was completed, the solution was filtered and washed with deionized water until the pH of the filtrate was 6.7. Then, the acid-treated diatomaceous earth was added to 1 mol / L sodium hydroxide solution at a material-to-liquid ratio of 1:7.5 (g / g). The solution was stirred at 85 °C for 1.5 h to undergo an alkaline dissolution reaction (dissolving amorphous silica and some impurities, exposing more silanol groups -Si-OH). After the reaction was completed, the solution was filtered and washed with deionized water until the pH of the filtrate was 6.7. The solution was dried at 105 °C for 5 h to obtain activated diatomaceous earth.

[0108] (2) Activated diatomaceous earth was dispersed in a 90% ethanol aqueous solution at a material-to-liquid ratio of 1:11.5 g / g, KH-550 was added, and a silanization reaction was carried out at 65℃ for 5 h (the mass ratio of KH-550 to activated diatomaceous earth was 0.2:1; the ethoxy group of KH-550 dehydrated and condensed with the silanol group on the surface of diatomaceous earth to form Si-O-Si covalent bonds, and amino-NH2 was introduced at the same time). After the reaction was completed, the mixture was filtered, washed three times with anhydrous ethanol, and dried at 55℃ for 6 h to obtain aminated diatomaceous earth.

[0109] (3) Aminated diatomaceous earth was dispersed in deionized water at a material-to-liquid ratio of 1:15 g / g, and tannic acid aqueous solution was added (the mass ratio of tannic acid to aminated diatomaceous earth was 0.4:1). The pH was adjusted to 8.5 with sodium bicarbonate. The mixture was stirred at 200 rpm at 55°C while air was introduced (flow rate 0.3 L / min) and reacted for 10 h (tannic acid was self-oxidized in weakly alkaline air to generate quinone intermediates, which underwent Michael addition reaction and Schiff base reaction with the amino groups on the surface of diatomaceous earth, thus covalently grafting tannic acid onto the surface of diatomaceous earth). After the reaction was completed, the mixture was filtered, washed 3 times with deionized water and 2 times with anhydrous ethanol, and then freeze-dried at -20°C and 0.3 mbar for 36 h (to avoid thermal oxidation and deactivation of phenolic hydroxyl groups) to obtain modified diatomaceous earth filter aid.

[0110] Preparation Example 3-3

[0111] This preparation example provides a modified diatomaceous earth filter aid, the preparation method of which is as follows:

[0112] (1) Add diatomaceous earth (particle size 50 μm) to 1 mol / L hydrochloric acid solution at a material-to-liquid ratio of 1:10. Stir at 300 rpm for 2 h in a constant temperature water bath at 90℃ to undergo acid dissolution reaction (dissolving impurities such as carbonates and metal oxides). After the reaction is completed, filter and wash with deionized water until the pH of the filtrate is 7.0. Then add the acid-treated diatomaceous earth to 1 mol / L sodium hydroxide solution at a material-to-liquid ratio of 1:10 (g / g). Stir at 90℃ for 2 h to undergo alkali dissolution reaction (dissolving amorphous silica and some impurities, exposing more silanol groups -Si-OH). After the reaction is completed, filter and wash with deionized water until the pH of the filtrate is 7.0. Dry at 110℃ for 6 h to obtain activated diatomaceous earth.

[0113] (2) Activated diatomaceous earth was dispersed in a 90% ethanol aqueous solution at a material-to-liquid ratio of 1:15 g / g, KH-550 was added, and a silanization reaction was carried out at 70℃ for 6 h (the mass ratio of KH-550 to activated diatomaceous earth was 0.3:1; the ethoxy group of KH-550 dehydrated and condensed with the silanol group on the surface of diatomaceous earth to form Si-O-Si covalent bonds, and amino-NH2 was introduced at the same time). After the reaction was completed, the mixture was filtered, washed three times with anhydrous ethanol, and dried at 60℃ for 8 h to obtain aminated diatomaceous earth.

[0114] (3) Aminated diatomaceous earth was dispersed in deionized water at a material-to-liquid ratio of 1:20 g / g, and tannic acid aqueous solution was added (the mass ratio of tannic acid to aminated diatomaceous earth was 0.5:1). The pH was adjusted to 9.0 with sodium bicarbonate. The mixture was stirred at 250 rpm at 60°C while air was introduced (flow rate 0.5 L / min) and reacted for 12 h (tannic acid was self-oxidized in weakly alkaline air to generate quinone intermediates, which underwent Michael addition reaction and Schiff base reaction with the amino groups on the surface of diatomaceous earth, thus covalently grafting tannic acid onto the surface of diatomaceous earth). After the reaction was completed, the mixture was filtered, washed 3 times with deionized water and 2 times with anhydrous ethanol, and then freeze-dried at -20°C and 0.5 mbar for 48 h (to avoid thermal oxidation and deactivation of phenolic hydroxyl groups) to obtain modified diatomaceous earth filter aid.

[0115] Example 1

[0116] This embodiment provides a method for preparing Ganoderma lucidum spore oil, including the following steps:

[0117] (1) Pre-adsorption of raw materials: Ganoderma lucidum spore powder raw material was mixed with modified chitosan microspheres prepared in Preparation Example 1-1 and loaded into the extraction vessel of a supercritical CO2 extraction device. Dynamic extraction was performed for 1 hour under the conditions of extraction pressure of 12 MPa, extraction temperature of 35 °C and CO2 flow rate of 15 L / h. After extraction, the pressure was reduced to atmospheric pressure and the modified chitosan microspheres were separated and removed by a 10-mesh vibrating sieve. The deplasticized Ganoderma lucidum spore powder was collected. The mass ratio of modified chitosan microspheres to Ganoderma lucidum spore powder raw material was 100:6.

[0118] (2) Supercritical extraction: The deplasticized Ganoderma lucidum spore powder was loaded into a supercritical extraction vessel. After static extraction for 30 min under the conditions of extraction pressure of 25 MPa, extraction temperature of 40℃ and CO2 flow rate of 20 L / h, dynamic extraction was performed for 2 h. The crude Ganoderma lucidum spore oil in the separation vessel was collected.

[0119] (3) Adsorption purification: Add a composite adsorbent (total addition amount is 2% of the crude Ganoderma lucidum spore oil) to the crude Ganoderma lucidum spore oil. The composite adsorbent is a mixture of the modified activated carbon adsorbent prepared in Preparation Example 2-1 and the modified diatomaceous earth filter aid prepared in Preparation Example 3-1 at a mass ratio of 3:1. Stir and adsorb at 30°C for 1 hour, and then remove the plasticizer from the Ganoderma lucidum spore oil by plate and frame filtration.

[0120] Example 2

[0121] This embodiment provides a method for preparing Ganoderma lucidum spore oil, including the following steps:

[0122] (1) Pre-adsorption of raw materials: Ganoderma lucidum spore powder raw material was mixed with modified chitosan microspheres prepared in Preparation Examples 1-2 and loaded into the extraction vessel of a supercritical CO2 extraction device. Dynamic extraction was carried out for 1.5 h under the conditions of extraction pressure of 15 MPa, extraction temperature of 40 °C and CO2 flow rate of 20 L / h. After extraction, the pressure was reduced to atmospheric pressure and the modified chitosan microspheres were separated and removed by a 10-mesh vibrating sieve. The deplasticized Ganoderma lucidum spore powder was collected. The mass ratio of modified chitosan microspheres to Ganoderma lucidum spore powder raw material was 100:7.

[0123] (2) Supercritical extraction: The deplasticized Ganoderma lucidum spore powder was loaded into a supercritical extraction vessel. After static extraction for 45 min under the conditions of extraction pressure of 30 MPa, extraction temperature of 45℃ and CO2 flow rate of 25 L / h, dynamic extraction was performed for 2.5 h. The crude Ganoderma lucidum spore oil in the separation vessel was collected.

[0124] (3) Adsorption purification: Add a composite adsorbent (total amount is 3% of the crude Ganoderma lucidum spore oil) to the crude Ganoderma lucidum spore oil. The composite adsorbent is a mixture of the modified activated carbon adsorbent prepared in Preparation Example 2-2 and the modified diatomaceous earth filter aid prepared in Preparation Example 3-2 at a mass ratio of 3.5:1. Stir and adsorb at 35°C for 2 hours, and then remove the plasticizer from the Ganoderma lucidum spore oil by plate and frame filtration.

[0125] Example 3

[0126] This embodiment provides a method for preparing Ganoderma lucidum spore oil, including the following steps:

[0127] (1) Pre-adsorption of raw materials: Ganoderma lucidum spore powder raw material was mixed with modified chitosan microspheres prepared in Preparation Examples 1-3 and loaded into the extraction vessel of a supercritical CO2 extraction device. Dynamic extraction was carried out for 2 hours under the conditions of extraction pressure of 18 MPa, extraction temperature of 45℃ and CO2 flow rate of 25 L / h. After extraction, the pressure was reduced to atmospheric pressure and the modified chitosan microspheres were separated and removed by a 10-mesh vibrating sieve. The deplasticized Ganoderma lucidum spore powder was collected. The mass ratio of modified chitosan microspheres to Ganoderma lucidum spore powder raw material was 100:8.

[0128] (2) Supercritical extraction: The deplasticized Ganoderma lucidum spore powder was loaded into a supercritical extraction vessel. After static extraction for 60 min under the conditions of extraction pressure of 35 MPa, extraction temperature of 50℃ and CO2 flow rate of 30 L / h, dynamic extraction was performed for 3 h. The crude Ganoderma lucidum spore oil in the separation vessel was collected.

[0129] (3) Adsorption purification: Add a composite adsorbent (total amount is 4% of the crude Ganoderma lucidum spore oil) to the crude Ganoderma lucidum spore oil. The composite adsorbent is a mixture of the modified activated carbon adsorbent prepared in Preparation Example 2-3 and the modified diatomaceous earth filter aid prepared in Preparation Example 3-3 at a mass ratio of 4:1. Stir and adsorb at 40°C for 3 hours, and then remove the plasticizer from the Ganoderma lucidum spore oil by plate and frame filtration.

[0130] Comparative Example 1

[0131] This comparative example provides a method for preparing Ganoderma lucidum spore oil, which differs from Example 1 only in that the modified chitosan microspheres of Preparation Example 1-1 are replaced with unmodified chitosan by an equal mass, while other conditions remain unchanged.

[0132] Comparative Example 2

[0133] This comparative example provides a method for preparing Ganoderma lucidum spore oil, which differs from Example 1 only in that the modified activated carbon adsorbent in Preparation Example 2-1 is replaced by an equal mass of unmodified activated carbon (particle size 0.5 mm), while other conditions remain unchanged.

[0134] Comparative Example 3

[0135] This comparative example provides a method for preparing Ganoderma lucidum spore oil, which differs from Example 1 only in that the modified diatomaceous earth filter aid in Preparation Example 3-1 is replaced by an equal mass of unmodified diatomaceous earth (particle size 10 μm), while other conditions remain unchanged.

[0136] Comparative Example 4

[0137] This comparative example provides a method for preparing Ganoderma lucidum spore oil, which differs from Example 1 only in that the composite adsorbent is replaced with the single modified activated carbon adsorbent of Preparation Example 2-1, and the amount of composite adsorbent used remains unchanged.

[0138] Comparative Example 5

[0139] This comparative example provides a method for preparing Ganoderma lucidum spore oil, which differs from Example 1 only in that the composite adsorbent is replaced with the single modified diatomaceous earth filter aid of Preparation Example 3-1, and the amount of composite adsorbent used remains unchanged.

[0140] Test case

[0141] The Ganoderma lucidum spore oil products obtained in Examples 1-3 and Comparative Examples 1-5 were tested for the following indicators:

[0142] (1) Total residual amount of plasticizers: The total residual amount (mg / kg) of 16 phthalate plasticizers in Ganoderma lucidum spore oil was detected by the test method in GB5009.271-2016.

[0143] (2) Ganoderma triterpenoid content: The content (%) of Ganoderma triterpenoids in Ganoderma spore oil was determined by the test method in the Pharmacopoeia of the People's Republic of China.

[0144] (3) Acid value: The acid value (mg KOH / g) of Ganoderma lucidum spore oil was determined by the test method in GB5009.229-2025.

[0145] The results are shown in Table 1.

[0146] Table 1

[0147]

[0148] As shown in Table 1, compared with the comparative method, the total residual amount of 16 plasticizers in the Ganoderma lucidum spore oil prepared by the method of this invention is significantly reduced, indicating that the residual amount of phthalate plasticizers in the Ganoderma lucidum spore oil is low, the removal process is highly efficient, and the product has good safety. The content of Ganoderma lucidum triterpenes is significantly higher than that in the comparative method, indicating that this process has a high retention rate of the main active ingredients (Ganoderma lucidum triterpenes) in the Ganoderma lucidum spore oil, and the deplasticization process does not cause significant loss of active ingredients, resulting in high health value. The acid value is significantly lower than that in the comparative method, indicating a low content of free fatty acids in the oil, mild rancidity, and high freshness and stability. Therefore, the preparation method of Ganoderma lucidum spore oil provided by this invention has a broader market prospect and is more suitable for promotion.

[0149] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0150] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0151] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A method for preparing ganoderma spore oil with reduced plasticizer residue, characterized in that, The method for preparing Ganoderma lucidum spore oil includes the following steps: (1) Pre-adsorption of raw materials: Ganoderma lucidum spore powder raw material is mixed with modified chitosan microspheres and subjected to supercritical CO2 extraction. After extraction, the modified chitosan microspheres are separated and removed, and the deplasticized Ganoderma lucidum spore powder is collected. (2) Supercritical extraction: The deplasticized Ganoderma lucidum spore powder was subjected to supercritical CO2 extraction to collect crude Ganoderma lucidum spore oil; (3) Adsorption purification: Ganoderma lucidum spore oil crude product is mixed with modified activated carbon and modified diatomaceous earth, and adsorption treatment is carried out. After filtration, Ganoderma lucidum spore oil is obtained.

2. The ganoderma lucidum spore oil preparation method according to claim 1, characterized in that, The mass ratio of the modified chitosan microspheres to the Ganoderma lucidum spore powder raw material in step (1) is 100:(1-15); Preferably, the supercritical CO2 extraction process in step (1) is carried out under the conditions of extraction pressure of 10-20 MPa, extraction temperature of 35-45℃, and CO2 flow rate of 15-25 L / h, with dynamic extraction for 1-4 h. Preferably, the separation in step (1) is performed using a 10-20 mesh vibrating screen.

3. The method of claim 1 or 2, wherein the Ganoderma lucidum spore oil is prepared by the method comprising the steps of: The particle size of the modified chitosan microspheres in step (1) is 0.5-1.5 mm; Preferably, the method for preparing the modified chitosan microspheres includes: Chitosan was treated with nano-silica sol, then mixed and emulsified with β-cyclodextrin and liquid paraffin containing Span-80. A crosslinking agent was then added to carry out the reaction. After the reaction was completed, the microspheres were collected by centrifugation, washed, dried, and sieved to obtain the modified chitosan microspheres.

4. The method of claim 3, wherein the Ganoderma lucidum spore oil is prepared by the steps of: The chitosan is treated with nano-silica sol as follows: chitosan is dissolved in an aqueous acetic acid solution with a volume fraction of 1.0-2.0% at a mass ratio of 1:(50-100). The chitosan acetic acid solution is added to the pre-ultrasonic dispersed nano-silica sol and stirred at 200-400 rpm for 30-60 min at 20-30℃. Preferably, the mass ratio of the nano-silica to chitosan is 1:(5-10); Preferably, the mass ratio of nano-silica to deionized water in the nano-silica sol is 1:(5-10); Preferably, the method of mixing and emulsifying is as follows: chitosan treated with nano-silica sol is mixed with β-cyclodextrin and stirred at 30-40°C for 1-2 h to form a composite aqueous phase, which is then mixed with liquid paraffin containing Span-80 as the oil phase, and emulsified at 8000-12000 rpm for 10-20 min to form a W / O type emulsion; Preferably, the mass ratio of β-cyclodextrin to chitosan is 1:(2-5); Preferably, the volume ratio of Span-80 to liquid paraffin is 1:(20-50); Preferably, the volume ratio of the aqueous phase to the oil phase is 1:(3-5); Preferably, the crosslinking agent is a glutaraldehyde aqueous solution with a mass fraction of 23-27%; Preferably, the mass ratio of glutaraldehyde to chitosan is (0.3-0.9):1; Preferably, the reaction is carried out at 40-50°C for 3-5 hours.

5. The method for preparing Ganoderma lucidum spore oil according to claim 1, characterized in that, In step (2), the supercritical CO2 extraction process involves static extraction for 20-70 min under the conditions of extraction pressure of 25-35 MPa, extraction temperature of 40-50℃, and CO2 flow rate of 20-30 L / h, followed by dynamic extraction for 1-4 h.

6. The method for preparing Ganoderma lucidum spore oil according to claim 1, characterized in that, The mass ratio of modified activated carbon to modified diatomaceous earth in step (3) is (2-5):1; Preferably, the total amount of modified activated carbon and modified diatomaceous earth added is 1-5% of the crude weight of Ganoderma lucidum spore oil; Preferably, the adsorption treatment in step (3) is carried out at 30-40℃ for 1-3 h; Preferably, the filtration in step (3) is performed using a plate and frame filter press.

7. The method for preparing Ganoderma lucidum spore oil according to any one of claims 1-6, characterized in that, The method for preparing the modified activated carbon in step (3) includes: Activated carbon was mixed with nitric acid solution for oxidation, washed, and dried to obtain oxidized activated carbon; the oxidized activated carbon was dispersed in an aqueous ethanol solution and mixed with a silane coupling agent for silanization, filtered, washed, and dried to obtain aminated activated carbon; the aminated activated carbon was dispersed in deionized water and mixed with citric acid and 1-ethyl-3-(2-methylaminopropyl)carbodiimide hydrochloride for amidation, filtered, washed, and dried to obtain the modified activated carbon.

8. The method for preparing Ganoderma lucidum spore oil according to claim 7, characterized in that, The activated carbon has a particle size of 0.5-1.0 mm; Preferably, the mass fraction of the nitric acid solution is 10-15%, and the mass ratio of the activated carbon to the nitric acid solution is 1:(5-10). Preferably, the oxidation reaction is carried out at 70-80°C for 2-3 hours; Preferably, the mass ratio of the oxidizing activated carbon to the ethanol aqueous solution is 1:(8-15), and the volume fraction of the ethanol aqueous solution is 85-95%. Preferably, the silane coupling agent is KH-550, and the mass ratio of KH-550 to activated carbon is (0.1-0.3):1; Preferably, the silanization reaction is carried out at 60-70°C for 4-6 hours; Preferably, the amidation reaction is carried out at pH 5.0-6.0 and 25-35°C for 4-8 h.

9. The method for preparing Ganoderma lucidum spore oil according to any one of claims 1-8, characterized in that, The preparation method of the modified diatomite in step (3) includes: Diatomaceous earth is mixed with an acid solution for acid treatment, and then the acid-treated diatomaceous earth is mixed with an alkaline solution for alkali dissolution reaction. After filtration, washing, and drying, activated diatomaceous earth is obtained. The activated diatomaceous earth is dispersed in an ethanol aqueous solution and mixed with a silane coupling agent for silanization reaction. After filtration, washing, and drying, aminated diatomaceous earth is obtained. The aminated diatomaceous earth is dispersed in deionized water and mixed with tannic acid for reaction. After filtration, washing, and drying, the modified diatomaceous earth is obtained.

10. The method for preparing Ganoderma lucidum spore oil according to claim 9, characterized in that, The particle size of the diatomaceous earth is 10-50 μm; Preferably, the acid solution is a 0.5-1.5 mol / L hydrochloric acid solution, and the mass ratio of the diatomaceous earth to the acid solution is 1:(5-10); Preferably, the acid treatment is carried out by stirring at 80-90°C for 1-2 hours; Preferably, the alkaline solution is a 0.5-1.5 mol / L sodium hydroxide solution, and the mass ratio of the acid-treated diatomaceous earth to the alkaline solution is 1:(5-10). Preferably, the alkali dissolution reaction is carried out by stirring at 80-90°C for 1-2 hours; Preferably, the volume fraction of the ethanol aqueous solution is 85-95%, and the mass ratio of the activated diatomaceous earth to the ethanol aqueous solution is 1:(8-15). Preferably, the silane coupling agent is KH-550, and the mass ratio of KH-550 to activated diatomaceous earth is (0.1-0.3):1; Preferably, the silanization reaction is carried out at 60-70°C for 4-6 hours; Preferably, the mass ratio of the aminated diatomaceous earth to deionized water is 1:(10-20); Preferably, the reaction with tannic acid is carried out at pH=8.0-9.0 and 50-60℃ for 8-12 h, and the mass ratio of tannic acid to aminated diatomaceous earth is (0.3-0.5):1.

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