Purification method of plasticizer-free ganoderma lucidum spore oil

Abstract

The invention relates to the technical field of deep processing of ganoderma lucidum spore oil, and discloses a purification method of plasticizer-free ganoderma lucidum spore oil, which comprises the following steps: raw material pretreatment, supercritical fluid extraction, analytical separation and post-treatment. In the supercritical fluid extraction step, supercritical carbon dioxide fluid with adjustable polarity is adopted, an entrainer is used in combination, a plasticizer is preferentially removed in the extraction process through the selective dissolution characteristic of the fluid to plasticizer molecules, meanwhile, the extraction interference on active ingredients in the ganoderma lucidum spore oil is minimized, and the extraction efficiency is improved. Therefore, the biological activity of the product is protected while the plasticizer is removed; meanwhile, the selective separation effect is further optimized through parameter control and dynamic adjustment in the extraction process, the loss of active ingredients caused by improper conditions in a traditional method is avoided, and the accuracy and reliability of the purification process are improved.

Description

Technical Field

[0001] This invention relates to the field of deep processing technology of Ganoderma lucidum spore oil, specifically a purification method for Ganoderma lucidum spore oil without plasticizers. Background Technology

[0002] Ganoderma lucidum spore oil is an oily lipid extracted from broken-cell Ganoderma lucidum spores using supercritical carbon dioxide fluid extraction technology. Its main components are triterpenoids, unsaturated fatty acids, and ganoderic acid, and it appears as a yellow, transparent liquid. The production process includes deep processing involving cell wall breaking and supercritical extraction, and it is believed to enhance immunity and regulate blood lipids.

[0003] Currently, when using traditional physical and chemical methods to purify and remove plasticizers from Ganoderma lucidum spore oil, it is difficult to selectively remove plasticizers because the physicochemical properties of plasticizers are similar to those of some effective components in Ganoderma lucidum spore oil. When separation conditions are not properly controlled, it will cause the loss of active ingredients, and it will be impossible to maintain the core efficacy of the product while ensuring the plasticizer removal effect.

[0004] Therefore, a method for purifying Ganoderma lucidum spore oil without plasticizers is proposed to solve the above problems. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a method for purifying Ganoderma lucidum spore oil without plasticizers, which solves the problem mentioned in the background art of loss of active ingredients and inability to maintain the core efficacy of the product while ensuring plasticizer removal.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a method for purifying Ganoderma lucidum spore oil without plasticizers, comprising the following steps: Step 1: Raw material pretreatment. Select raw Ganoderma lucidum spore oil, degumm it and filter it to remove impurities and some moisture; Step 2: Supercritical fluid extraction. The pretreated Ganoderma lucidum spore oil is mixed with supercritical carbon dioxide fluid and extracted under specific pressure and temperature to selectively dissolve and remove plasticizer components. Step 3: Separation and analysis. The extracted supercritical fluid is subjected to reduced pressure and temperature control to separate the plasticizer from the carbon dioxide, and the purified Ganoderma lucidum spore oil is collected. Step 4: Post-processing. The separated Ganoderma lucidum spore oil is desolventized and dried to ensure no solvent residue, thus obtaining high-purity, plasticizer-free Ganoderma lucidum spore oil. In the supercritical fluid extraction step, the pressure of supercritical carbon dioxide is set to 20-35 MPa, the temperature is set to 35-60°C, and the extraction time is 1-4 hours; in the analytical separation step, the pressure is reduced to 5-10 MPa and the temperature is adjusted to 25-40°C.

[0007] Preferably, the raw material pretreatment in step one includes the following steps: The raw Ganoderma lucidum spore oil is heated to 40-60°C, and a degumming agent, either phosphoric acid or citric acid, is added at a rate of 0.1-0.5% of the weight of the Ganoderma lucidum spore oil. The mixture is stirred and mixed for 30-60 minutes, and then centrifuged at 3000-5000 r / min for 10-20 minutes to separate the colloids and impurities. Subsequently, the centrifuged oil phase is filtered through a microporous membrane with a pore size of 0.2-0.5 μm and a filtration pressure of 0.1-0.3 MPa to remove residual particles and moisture, thus obtaining pretreated Ganoderma lucidum spore oil.

[0008] Preferably, the step of filtering through a microporous membrane specifically includes: First, a pre-filter with a pore size of 5-10 μm is used to coarsely filter the degummed oil phase at a filtration pressure of 0.05-0.1 MPa to remove larger particulate impurities. Then, a fine filter with a pore size of 0.2-0.5 μm is used for fine filtration at a filtration pressure of 0.1-0.3 MPa to ensure that the oil phase is clear.

[0009] Preferably, the supercritical fluid extraction in step two uses an entrainer, which is at least one of ethanol and methanol, and the amount added is 5-15% of the weight of supercritical carbon dioxide. The entrainer is premixed with carbon dioxide before extraction. The mixing method is to inject the entrainer into the carbon dioxide stream at a flow rate of 1-3 mL / min using a high-pressure pump, and mix it in a static mixer at a pressure of 10-20 MPa for 5-10 minutes to form a uniform supercritical fluid system. The extraction process is carried out in an extraction vessel, and the loading of Ganoderma lucidum spore oil is 30-60% of the volume of the extraction vessel, and the carbon dioxide flow rate is 10-30 L / h.

[0010] Preferably, when the entrainer is ethanol, its concentration is above 95%, and the mixing ratio of ethanol and carbon dioxide is controlled by a mass flow meter. The amount of ethanol added is 8-12%. The extraction vessel is equipped with a temperature control system with a temperature control accuracy of ±1°C and a pressure control accuracy of ±0.5MPa. During the extraction process, samples are taken every 20-30 minutes to detect the plasticizer content until the plasticizer residue is below 0.1mg / kg, at which point the extraction is terminated.

[0011] Preferably, the supercritical carbon dioxide fluid in step two is obtained through a multi-stage extraction method, including a first-stage extraction and a second-stage extraction. The first-stage extraction pressure is 20-25 MPa and the temperature is 35-45°C, removing most of the weakly polar plasticizers. The second-stage extraction pressure is 30-35 MPa and the temperature is 50-60°C, removing the residual strongly polar plasticizers. The extraction time for each stage is 1-2 hours, and the carbon dioxide flow rate is 15-25 L / h. The two stages are switched by a valve to avoid cross-contamination. The multi-stage extraction system consists of extraction vessels connected in series, each with the same volume. The Ganoderma lucidum spore oil is processed in batches sequentially in each extraction vessel.

[0012] Preferably, the supercritical fluid extraction in step two is carried out under stirring conditions, with the stirring speed set to 100-300 r / min; during the extraction process, the density of the carbon dioxide fluid is adjusted in real time and controlled within the range of 0.5-0.8 g / mL, and the extraction time is dynamically adjusted according to the oil loading amount, with an extraction time of 0.5-1 hour per kilogram of oil.

[0013] Preferably, the separation in step three is achieved through multi-stage decompression, including primary separation and secondary separation. Primary separation is carried out in a separation vessel, where the pressure is reduced from the extraction pressure to 10-15 MPa and the temperature is maintained at 35-45°C, causing most of the carbon dioxide to vaporize and carry away the plasticizer. Secondary separation is carried out in another separation vessel, where the pressure is further reduced to 5-8 MPa and the temperature is adjusted to 25-30°C, causing the remaining plasticizer to precipitate. The separated carbon dioxide is recovered through a condenser and recycled. During the separation process, an ultrasonic auxiliary device is installed in the separation vessel, with an ultrasonic frequency of 20-40 kHz, a power of 100-300 W, and an action time of 10-20 minutes.

[0014] Preferably, the analytical separation step in step three further includes adsorption treatment, in which an adsorbent is added to the separation vessel to further remove trace plasticizers; the adsorbent is at least one of activated carbon, silica gel or molecular sieve, and the amount added is 1-3% of the weight of Ganoderma lucidum spore oil. The adsorbent is pre-loaded in the filter screen of the separation vessel before analytical separation, and the contact time with the oil is 30-60 minutes. After adsorption, the adsorbent is separated by a filter press at a pressure of 0.2-0.5 MPa to ensure the oil is clear. The adsorbent can be regenerated and reused by calcining at 300-400°C for 1-2 hours.

[0015] Preferably, the post-processing in step four includes solvent removal and drying. Solvent removal is performed using a vacuum solvent removal device with a vacuum degree set to 0.05-0.1 MPa, a temperature of 50-60°C, and a solvent removal time of 1-2 hours to remove residual carbon dioxide and entrainers. Drying is performed using either spray drying or vacuum drying. During spray drying, the inlet temperature is 150-200°C, the outlet temperature is 80-100°C, and the atomization pressure is 0.3-0.6 MPa. During vacuum drying, the vacuum degree is 0.01-0.05 MPa, the temperature is 40-50°C, and the drying time is 2-3 hours. After drying, the moisture content of the Ganoderma lucidum spore oil is less than 0.1%, and it is stored in nitrogen packaging.

[0016] Compared with the prior art, the present invention provides a method for purifying Ganoderma lucidum spore oil without plasticizers, which has the following beneficial effects: 1. In this invention, the supercritical fluid extraction step uses supercritical carbon dioxide fluid with adjustable polarity, combined with the use of an entrainer. Utilizing the selective solubility characteristics of the fluid for plasticizer molecules, the extraction process prioritizes the removal of plasticizers while minimizing interference with the extraction of active ingredients in Ganoderma lucidum spore oil. This achieves the removal of plasticizers while protecting the bioactivity of the product. Furthermore, parameter control and dynamic adjustment during the extraction process further optimize the selective separation effect, avoiding the loss of active ingredients due to improper conditions in traditional methods, and improving the accuracy and reliability of the purification process.

[0017] 2. In this invention, the multi-stage extraction system extracts weakly polar and strongly polar plasticizers in stages by setting different pressure and temperature parameters. By utilizing the targeted conditions of each extraction stage, it achieves simultaneous removal of plasticizers with different properties, thereby reducing plasticizer residues and improving the purity and safety of the product. In addition, the multi-stage extraction combined with stirring improves fluid turbulence and mass transfer efficiency, ensuring the removal of mixed plasticizers and avoiding the removal blind spots under single technical parameters.

[0018] 3. In this invention, an adsorption treatment process is introduced in the analytical separation stage. Through the adsorption of activated carbon, silica gel or molecular sieve, trace plasticizer molecules remaining in the oil phase are captured and retained to remove the target impurities. Subsequently, in the post-treatment stage, vacuum desolventizing and controlled drying processes are used to remove trace entrainers and moisture in the system under specific temperature and pressure conditions, reducing the risk of solvent residue. Detailed Implementation

[0019] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0020] Example 1: A method for purifying Ganoderma lucidum spore oil without plasticizers, comprising the following steps: Step 1: Raw material pretreatment. Select raw Ganoderma lucidum spore oil, degumm it and filter it to remove impurities and some moisture; Step 2: Supercritical fluid extraction. The pretreated Ganoderma lucidum spore oil is mixed with supercritical carbon dioxide fluid and extracted under specific pressure and temperature to selectively dissolve and remove plasticizer components. Step 3: Separation and analysis. The extracted supercritical fluid is subjected to reduced pressure and temperature control to separate the plasticizer from the carbon dioxide, and the purified Ganoderma lucidum spore oil is collected. Step 4: Post-processing. The separated Ganoderma lucidum spore oil is desolventized and dried to ensure no solvent residue, thus obtaining high-purity, plasticizer-free Ganoderma lucidum spore oil. In the supercritical fluid extraction step, the pressure of supercritical carbon dioxide is set to 20 MPa, the temperature is set to 35°C, and the extraction time is 1 hour; in the desorption and separation step, the pressure is reduced to 5 MPa and the temperature is adjusted to 25°C.

[0021] The raw material pretreatment in step one includes the following steps: The raw Ganoderma lucidum spore oil was heated to 40°C, and a degumming agent, phosphoric acid, was added at 0.1% of the weight of the Ganoderma lucidum spore oil. The mixture was stirred and mixed for 30 minutes, and then centrifuged at 3000 r / min for 10 minutes to separate the colloids and impurities. Subsequently, the centrifuged oil phase was filtered through a microporous membrane with a pore size of 0.2 μm and a filtration pressure of 0.1 MPa to remove residual particles and moisture, thus obtaining pretreated Ganoderma lucidum spore oil.

[0022] The specific steps involved in filtration using a microporous membrane include: First, a pre-filter with a pore size of 5 μm is used to coarsely filter the degummed oil phase at a filtration pressure of 0.05 MPa to remove larger particulate impurities. Then, a fine filter with a pore size of 0.2 μm is used for fine filtration at a filtration pressure of 0.1 MPa to ensure that the oil phase is clear.

[0023] In step two, supercritical fluid extraction uses an entrainer, which is at least one of ethanol and methanol, and the amount added is 5% of the weight of supercritical carbon dioxide. The entrainer is premixed with carbon dioxide before extraction. The mixing method is to inject the entrainer into the carbon dioxide stream at a flow rate of 1 mL / min using a high-pressure pump, and mix it in a static mixer at a pressure of 10 MPa for 5 minutes to form a homogeneous supercritical fluid system. The extraction process is carried out in an extraction vessel, and the loading of Ganoderma lucidum spore oil is 30% of the volume of the extraction vessel, and the carbon dioxide flow rate is 10 L / h.

[0024] When the entrainer is ethanol, its concentration is above 95%, and the mixing ratio of ethanol and carbon dioxide is controlled by a mass flow meter. The amount of ethanol added is 8%. The extraction vessel is equipped with a temperature control system with a temperature control accuracy of ±1°C and a pressure control accuracy of ±0.5MPa. During the extraction process, samples are taken every 20 minutes to test the plasticizer content until the plasticizer content reaches the standard, and then the extraction is terminated.

[0025] In step two, the supercritical carbon dioxide fluid is extracted through a multi-stage extraction process, including a first-stage extraction and a second-stage extraction. The first-stage extraction pressure is 20 MPa and the temperature is 35°C, which removes most of the weakly polar plasticizers. The second-stage extraction pressure is 30 MPa and the temperature is 50°C, which removes the residual strongly polar plasticizers. The extraction time for each stage is 1 hour, and the carbon dioxide flow rate is 15 L / h. The two stages are switched by a valve to avoid cross-contamination. The multi-stage extraction system consists of extraction vessels connected in series, each with the same volume. The Ganoderma lucidum spore oil is processed in batches in each extraction vessel.

[0026] The supercritical fluid extraction in step two is carried out under stirring conditions, with the stirring speed set to 100 r / min. During the extraction process, the density of the carbon dioxide fluid is adjusted in real time and controlled at 0.5 g / mL. The extraction time is dynamically adjusted according to the oil loading, with an extraction time of 0.5 hours per kilogram of oil.

[0027] The separation in step three is achieved through multi-stage depressurization, including primary and secondary separation. Primary separation is carried out in a separation vessel, where the pressure is reduced from the extraction pressure to 10 MPa and the temperature is maintained at 35°C, causing most of the carbon dioxide to vaporize and carry away the plasticizer. Secondary separation is carried out in another separation vessel, where the pressure is further reduced to 5 MPa and the temperature is adjusted to 25°C, causing the remaining plasticizer to precipitate. The separated carbon dioxide is recovered through a condenser and recycled. During the separation process, an ultrasonic auxiliary device is installed in the separation vessel, with an ultrasonic frequency of 20 kHz, a power of 100 W, and an action time of 10 minutes.

[0028] The separation step in step three also includes adsorption treatment, in which an adsorbent is added to the separation vessel to further remove trace plasticizers. The adsorbent is activated carbon, and the amount added is 1% of the weight of Ganoderma lucidum spore oil. The adsorbent is pre-loaded in the filter screen of the separation vessel before desorption, and the contact time with the oil is 30 minutes. After adsorption, the adsorbent is separated by a filter press at a pressure of 0.2 MPa to ensure that the oil is clear. The adsorbent can be regenerated and reused. The regeneration method is to calcine at 300°C for 1 hour.

[0029] The post-processing in step four includes solvent removal and drying. Solvent removal is performed using a vacuum solvent removal device with a vacuum level of 0.05 MPa, a temperature of 50°C, and a solvent removal time of 1 hour to remove residual carbon dioxide and entrainers. Drying is performed using either spray drying or vacuum drying. During spray drying, the inlet temperature is 150°C, the outlet temperature is 80°C, and the atomization pressure is 0.3 MPa. During vacuum drying, the vacuum level is 0.01 MPa, the temperature is 40°C, and the drying time is 2 hours. After drying, the moisture content of the Ganoderma lucidum spore oil is less than 0.1%, and it is stored in nitrogen packaging.

[0030] Example 2: A method for purifying Ganoderma lucidum spore oil without plasticizers, comprising the following steps: Step 1: Raw material pretreatment. Select raw Ganoderma lucidum spore oil, degumm it and filter it to remove impurities and some moisture; Step 2: Supercritical fluid extraction. The pretreated Ganoderma lucidum spore oil is mixed with supercritical carbon dioxide fluid and extracted under specific pressure and temperature to selectively dissolve and remove plasticizer components. Step 3: Separation and analysis. The extracted supercritical fluid is subjected to reduced pressure and temperature control to separate the plasticizer from the carbon dioxide, and the purified Ganoderma lucidum spore oil is collected. Step 4: Post-processing. The separated Ganoderma lucidum spore oil is desolventized and dried to ensure no solvent residue, thus obtaining high-purity, plasticizer-free Ganoderma lucidum spore oil. In the supercritical fluid extraction step, the pressure of supercritical carbon dioxide was set to 25 MPa, the temperature to 45°C, and the extraction time to 2 hours; in the desorption and separation step, the pressure was reduced to 7 MPa and the temperature was adjusted to 35°C.

[0031] The raw material pretreatment in step one includes the following steps: The raw Ganoderma lucidum spore oil was heated to 50°C, and a degumming agent, phosphoric acid, was added at a rate of 0.3% of the weight of the Ganoderma lucidum spore oil. The mixture was stirred and mixed for 45 minutes, and then centrifuged at 4000 r / min for 15 minutes to separate the colloids and impurities. Subsequently, the centrifuged oil phase was filtered through a microporous membrane with a pore size of 0.3 μm and a filtration pressure of 0.2 MPa to remove residual particles and moisture, thus obtaining pretreated Ganoderma lucidum spore oil.

[0032] The specific steps involved in filtration using a microporous membrane include: First, a pre-filter with a pore size of 57 μm is used to coarsely filter the degummed oil phase at a filtration pressure of 0.07 MPa to remove larger particulate impurities. Then, a fine filter with a pore size of 0.3 μm is used for fine filtration at a filtration pressure of 0.2 MPa to ensure that the oil phase is clear.

[0033] In step two, supercritical fluid extraction uses an entrainer, which is at least one of ethanol and methanol, and the amount added is 10% of the weight of supercritical carbon dioxide. The entrainer is premixed with carbon dioxide before extraction. The mixing method is to inject the entrainer into the carbon dioxide stream at a flow rate of 2 mL / min using a high-pressure pump, and mix it in a static mixer at a pressure of 15 MPa for 7 minutes to form a homogeneous supercritical fluid system. The extraction process is carried out in an extraction vessel, with the loading of Ganoderma lucidum spore oil at 45% of the volume of the extraction vessel and the carbon dioxide flow rate at 20 L / h.

[0034] When the entrainer is ethanol, its concentration is above 95%, and the mixing ratio of ethanol and carbon dioxide is controlled by a mass flow meter. The amount of ethanol added is 10%. The extraction vessel is equipped with a temperature control system with a temperature control accuracy of ±1°C and a pressure control accuracy of ±0.5MPa. During the extraction process, samples are taken every 25 minutes to test the plasticizer content until the plasticizer content reaches the standard, and then the extraction is terminated.

[0035] In step two, the supercritical carbon dioxide fluid is extracted through a multi-stage extraction process, including a first-stage extraction and a second-stage extraction. The first-stage extraction pressure is 22 MPa and the temperature is 40°C, which removes most of the weakly polar plasticizers. The second-stage extraction pressure is 32 MPa and the temperature is 55°C, which removes the residual strongly polar plasticizers. The extraction time for each stage is 1.5 hours, and the carbon dioxide flow rate is 20 L / h. The two stages are switched by a valve to avoid cross-contamination. The multi-stage extraction system consists of extraction vessels connected in series, each with the same volume. The Ganoderma lucidum spore oil is processed in batches in each extraction vessel sequentially.

[0036] The supercritical fluid extraction in step two was carried out under stirring conditions, with the stirring speed set at 200 r / min. During the extraction process, the density of the carbon dioxide fluid was adjusted in real time and controlled at 0.7 g / mL. The extraction time was dynamically adjusted according to the amount of oil loaded, with an extraction time of 0.8 hours per kilogram of oil.

[0037] The separation in step three is achieved through multi-stage depressurization, including primary and secondary separation. Primary separation is carried out in a separation vessel, where the pressure is reduced from the extraction pressure to 12 MPa and the temperature is maintained at 40°C, causing most of the carbon dioxide to vaporize and carry away the plasticizer. Secondary separation is carried out in another separation vessel, where the pressure is further reduced to 7 MPa and the temperature is adjusted to 27°C, causing the remaining plasticizer to precipitate. The separated carbon dioxide is recovered through a condenser and recycled. During the separation process, an ultrasonic auxiliary device is installed in the separation vessel, with an ultrasonic frequency of 30 kHz, a power of 200 W, and an action time of 15 minutes.

[0038] The separation step in step three also includes adsorption treatment, in which an adsorbent is added to the separation vessel to further remove trace plasticizers. The adsorbent is activated carbon, and the amount added is 2% of the weight of Ganoderma lucidum spore oil. The adsorbent is pre-loaded in the filter screen of the separation vessel before desorption, and the contact time with the oil is 45 minutes. After adsorption, the adsorbent is separated by a filter press at a pressure of 0.3 MPa to ensure that the oil is clear. The adsorbent can be regenerated and reused. The regeneration method is to calcine at 350°C for 1.5 hours.

[0039] The post-processing in step four includes solvent removal and drying. Solvent removal is performed using a vacuum solvent removal device with a vacuum level of 0.07 MPa, a temperature of 55°C, and a solvent removal time of 1.5 hours to remove residual carbon dioxide and entrainers. Drying is performed using either spray drying or vacuum drying. During spray drying, the inlet temperature is 170°C, the outlet temperature is 90°C, and the atomization pressure is 0.5 MPa. During vacuum drying, the vacuum level is 0.03 MPa, the temperature is 45°C, and the drying time is 2.5 hours. After drying, the moisture content of the Ganoderma lucidum spore oil is less than 0.1%, and it is stored in nitrogen packaging.

[0040] Example 3: A method for purifying Ganoderma lucidum spore oil without plasticizers, comprising the following steps: Step 1: Raw material pretreatment. Select raw Ganoderma lucidum spore oil, degumm it and filter it to remove impurities and some moisture; Step 2: Supercritical fluid extraction. The pretreated Ganoderma lucidum spore oil is mixed with supercritical carbon dioxide fluid and extracted under specific pressure and temperature to selectively dissolve and remove plasticizer components. Step 3: Separation and analysis. The extracted supercritical fluid is subjected to reduced pressure and temperature control to separate the plasticizer from the carbon dioxide, and the purified Ganoderma lucidum spore oil is collected. Step 4: Post-processing. The separated Ganoderma lucidum spore oil is desolventized and dried to ensure no solvent residue, thus obtaining high-purity, plasticizer-free Ganoderma lucidum spore oil. In the supercritical fluid extraction step, the pressure of supercritical carbon dioxide was set to 35 MPa, the temperature to 60°C, and the extraction time to 4 hours; in the desorption and separation step, the pressure was reduced to 10 MPa and the temperature was adjusted to 40°C.

[0041] The raw material pretreatment in step one includes the following steps: The raw Ganoderma lucidum spore oil was heated to 60°C, and a degumming agent, phosphoric acid, was added at 0.5% of the weight of the Ganoderma lucidum spore oil. The mixture was stirred and mixed for 60 minutes, and then centrifuged at 5000 r / min for 20 minutes to separate the colloids and impurities. Subsequently, the centrifuged oil phase was filtered through a microporous membrane with a pore size of 0.5 μm and a filtration pressure of 0.3 MPa to remove residual particles and moisture, thus obtaining pretreated Ganoderma lucidum spore oil.

[0042] The specific steps involved in filtration using a microporous membrane include: First, a pre-filter with a pore size of 10 μm is used to coarsely filter the degummed oil phase at a filtration pressure of 0.1 MPa to remove larger particulate impurities. Then, a fine filter with a pore size of 0.5 μm is used for fine filtration at a filtration pressure of 0.3 MPa to ensure that the oil phase is clear.

[0043] In step two, supercritical fluid extraction uses an entrainer, which is at least one of ethanol and methanol, and the amount added is 15% of the weight of supercritical carbon dioxide. The entrainer is premixed with carbon dioxide before extraction. The mixing method is to inject the entrainer into the carbon dioxide stream at a flow rate of 3 mL / min using a high-pressure pump, and mix it in a static mixer at a pressure of 20 MPa for 10 minutes to form a homogeneous supercritical fluid system. The extraction process is carried out in an extraction vessel, with the loading of Ganoderma lucidum spore oil at 60% of the volume of the extraction vessel and the carbon dioxide flow rate at 30 L / h.

[0044] When the entrainer is ethanol, its concentration is above 95%, and the mixing ratio of ethanol and carbon dioxide is controlled by a mass flow meter. The amount of ethanol added is 12%. The extraction vessel is equipped with a temperature control system with a temperature control accuracy of ±1°C and a pressure control accuracy of ±0.5MPa. During the extraction process, samples are taken every 30 minutes to test the plasticizer content until the plasticizer content reaches the standard, and then the extraction is terminated.

[0045] In step two, the supercritical carbon dioxide fluid is extracted through a multi-stage extraction process, including a first-stage extraction and a second-stage extraction. The first-stage extraction pressure is 25 MPa and the temperature is 45°C, which removes most of the weakly polar plasticizers. The second-stage extraction pressure is 35 MPa and the temperature is 60°C, which removes the residual strongly polar plasticizers. Each stage of extraction takes 2 hours, and the carbon dioxide flow rate is 25 L / h. The two stages are switched by a valve to avoid cross-contamination. The multi-stage extraction system consists of extraction vessels connected in series, each with the same volume. The Ganoderma lucidum spore oil is processed in batches in each extraction vessel.

[0046] The supercritical fluid extraction in step two was carried out under stirring conditions, with the stirring speed set at 300 r / min. During the extraction process, the density of the carbon dioxide fluid was adjusted in real time and controlled at 0.8 g / mL. The extraction time was dynamically adjusted according to the amount of oil loaded, with an extraction time of 1 hour per kilogram of oil.

[0047] The separation in step three is achieved through multi-stage depressurization, including primary and secondary separation. Primary separation is carried out in a separation vessel, where the pressure is reduced from the extraction pressure to 15 MPa and the temperature is maintained at 45°C, causing most of the carbon dioxide to vaporize and carry away the plasticizer. Secondary separation is carried out in another separation vessel, where the pressure is further reduced to 8 MPa and the temperature is adjusted to 30°C, causing the remaining plasticizer to precipitate. The separated carbon dioxide is recovered through a condenser and recycled. During the separation process, an ultrasonic auxiliary device is installed in the separation vessel, with an ultrasonic frequency of 40 kHz, a power of 300 W, and an action time of 20 minutes.

[0048] The separation step in step three also includes adsorption treatment, in which an adsorbent is added to the separation vessel to further remove trace plasticizers. The adsorbent is activated carbon, and the amount added is 3% of the weight of Ganoderma lucidum spore oil. The adsorbent is pre-loaded in the filter screen of the separation vessel before desorption, and the contact time with the oil is 60 minutes. After adsorption, the adsorbent is separated by a filter press at a pressure of 0.5 MPa to ensure that the oil is clear. The adsorbent can be regenerated and reused. The regeneration method is to calcine at 400°C for 2 hours.

[0049] The post-processing in step four includes solvent removal and drying. Solvent removal is performed using a vacuum solvent removal device with a vacuum level of 0.1 MPa, a temperature of 60°C, and a solvent removal time of 2 hours to remove residual carbon dioxide and entrainers. Drying is performed using either spray drying or vacuum drying. During spray drying, the inlet temperature is 1200°C, the outlet temperature is 100°C, and the atomization pressure is 0.6 MPa. During vacuum drying, the vacuum level is 0.05 MPa, the temperature is 50°C, and the drying time is 3 hours. After drying, the moisture content of the Ganoderma lucidum spore oil is less than 0.1%, and it is stored in nitrogen packaging.

[0050] Comparative Example 1: This comparative example differs from Example 1 in that no entrainer was used during supercritical fluid extraction.

[0051] Comparative Example 2 differs from Example 1 in that it uses single-stage extraction instead of multi-stage extraction when performing supercritical fluid extraction.

[0052] Comparative Example 3 differs from Example 1 in that no adsorption treatment was performed during the analytical separation.

[0053] Comparative Example 4 differs from Example 1 in that degumming and filtration were not performed during the raw material pretreatment.

[0054] The plasticizer-free Ganoderma lucidum spore oils prepared in Examples 1-3 and Comparative Examples 1-4 were subjected to performance tests. The test items and test methods are as follows: For the determination of plasticizer residues, gas chromatography-mass spectrometry (GC-MS) was used with a DB-5MS column. The column temperature program was as follows: initial temperature 60℃, hold for 1 minute, increase to 220℃ at 20℃ / min, hold for 1 minute, increase to 250℃ at 5℃ / min, hold for 1 minute, then increase to 290℃ at 20℃ / min, hold for 7.5 minutes. The injection port temperature was 260℃, the ion source temperature was 230℃, and selected ion monitoring mode was used. Qualitative and quantitative analysis of common phthalate plasticizers, namely DBP, DEHP, and DINP, was performed. The content of active ingredients was tested using a high-performance liquid chromatograph with a C18 column and a methanol-water gradient elution mobile phase. The detection wavelength was selected as the maximum absorption wavelength of the characteristic active ingredients in Ganoderma lucidum spore oil. The content of characteristic active ingredients in Ganoderma lucidum spore oil before and after purification was determined by the external standard method. For the spore oil yield test, the mass of Ganoderma lucidum spore oil raw material before purification was weighed. After purification, the mass of the final plasticizer-free Ganoderma lucidum spore oil was collected and weighed, and the yield was calculated using the formula: mass of purified spore oil / mass of raw spore oil before purification × 100%. Solvent residue and moisture content were tested. Solvent residue was determined by headspace gas chromatography under the same chromatographic conditions as the plasticizer residue test, and the entrainer ethanol was tested. Moisture content was determined by the Karl Fischer method.

[0055] Table 1: Comprehensive Performance Test Data of Plasticizer-Free Ganoderma Spore Oil

[0056] By comparing and analyzing the data in Table 1, it can be seen that the plasticizer-free Ganoderma lucidum spore oil prepared using the processes in Examples 1-3 exhibits advantages in performance indicators compared to the products prepared using the processes in Comparative Examples 1-4. This indicates that the supercritical fluid extraction step, employing adjustable polarity supercritical carbon dioxide fluid and combined with the use of an entrainer, leverages the selective solubility characteristics of the fluid for plasticizer molecules to preferentially remove plasticizers during the extraction process, while minimizing interference with the extraction of active ingredients in the Ganoderma lucidum spore oil. This achieves the removal of plasticizers while protecting the bioactivity of the product. Furthermore, the parameter control and dynamic adjustment during the extraction process further optimize the selective separation effect, avoiding the loss of active ingredients due to improper conditions in traditional methods, and improving the accuracy and reliability of the purification process. The multi-stage extraction system extracts weakly and strongly polar plasticizers in stages by setting different pressure and temperature parameters. Utilizing the targeted conditions of each extraction stage, it achieves simultaneous removal of plasticizers with different properties, thereby reducing plasticizer residue and improving product purity and safety. Furthermore, the multi-stage extraction combined with stirring improves fluid turbulence and mass transfer efficiency, ensuring the removal of mixed plasticizers and avoiding the blind spots of removal under single technical parameters. An adsorption treatment process is introduced in the analytical separation stage, using activated carbon, silica gel, or molecular sieves to capture and retain trace plasticizer molecules remaining in the oil phase, achieving the removal of target impurities. Subsequently, the post-treatment stage employs vacuum desolventizing and controlled drying processes to remove trace entrainers and moisture in the system under specific temperature and pressure conditions, reducing the risk of solvent residue.

[0057] Table 2: Test data on plasticizer residues

[0058] Plasticizer residue is a key indicator for assessing the safety of edible oils. According to the data in Table 2, after using the novel purification process in Examples 1-3, the residues of the three common plasticizers, DBP, DEHP, and DINP, were all below the method detection limit (<0.01 mg / kg), and the total plasticizer residue was <0.03 mg / kg. In contrast, Comparative Examples 1-4, using the traditional purification method, showed plasticizer residues between 0.46 and 0.73 mg / kg. Experiments demonstrate that the supercritical CO2 extraction combined with molecular sieve adsorption purification process used in this invention can remove plasticizers from spore oil, ensuring product safety.

[0059] Table 3: Test data on retention rate of active ingredients

[0060] The retention rate of active ingredients is an important parameter for evaluating the quality of purification processes. Table 3 shows that in Examples 1-3, the retention rates of triterpenoids, ganoderic acids, ganoderic polysaccharides, and sterols, the main active ingredients, were all above 95%, with a total active ingredient retention rate of 96.9%-97.5%. In contrast, the total active ingredient retention rate in Comparative Examples 1-4 was only 80.6%-88.3%. This demonstrates that the present invention can maximize the retention of bioactive components in Ganoderma lucidum spore oil while removing impurities, ensuring that the product efficacy is not affected.

[0061] Table 4: Spore oil yield and acid value / peroxide value test

[0062] The spore oil yield and quality indicators reflect the efficiency of the purification process and the product quality. Table 4 shows that the spore oil yield of Examples 1-3 was 94.2%-95.3%, higher than the 90.2%-94.7% of Comparative Examples 1-4. Meanwhile, the acid value of the Example group (0.78-0.85 mg KOH / g) was lower than that of the Comparative Example group, and the color was lighter and the transparency was better. This indicates that the process of the present invention not only improves the extraction efficiency of spore oil but also improves the oxidative stability and sensory quality of the product, extending its shelf life.

[0063] Table 5: Solvent Residue and Microbiological Indicator Tests

[0064] Solvent residue and microbiological indicators are important parameters for evaluating the safety and hygiene of edible oils. Table 5 shows that in Examples 1-3, the residues of n-hexane and ethanol were below the method detection limit, the total solvent residue was <2 mg / kg, and the total bacterial count, mold and yeast count were all <10 CFU / g, with no detectable coliforms, indicating excellent microbiological indicators. In contrast, the solvent residues in Comparative Examples 1-4 ranged from 15.4-38.7 mg / kg, and the number of microorganisms were higher than in the Example groups. This indicates that the solvent-free purification process combined with low-temperature sterilization technology used in this invention can control solvent residue and microbial contamination, ensuring that the product meets food safety requirements.

[0065] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0066] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for purifying Ganoderma lucidum spore oil without plasticizers, characterized in that: Includes the following steps: Step 1: Raw material pretreatment. Select raw Ganoderma lucidum spore oil, degumm it and filter it to remove impurities and some moisture; Step 2: Supercritical fluid extraction. The pretreated Ganoderma lucidum spore oil is mixed with supercritical carbon dioxide fluid and extracted under specific pressure and temperature to selectively dissolve and remove plasticizer components. Step 3: Separation and analysis. The extracted supercritical fluid is subjected to reduced pressure and temperature control to separate the plasticizer from the carbon dioxide, and the purified Ganoderma lucidum spore oil is collected. Step 4: Post-processing. The separated Ganoderma lucidum spore oil is desolventized and dried to ensure no solvent residue, thus obtaining high-purity, plasticizer-free Ganoderma lucidum spore oil. In the supercritical fluid extraction step, the pressure of supercritical carbon dioxide is set to 20-35 MPa, the temperature is set to 35-60°C, and the extraction time is 1-4 hours; in the analytical separation step, the pressure is reduced to 5-10 MPa and the temperature is adjusted to 25-40°C.

2. The purification method for Ganoderma lucidum spore oil without plasticizers according to claim 1, characterized in that: The raw material pretreatment in step one includes the following steps: The raw Ganoderma lucidum spore oil is heated to 40-60°C, and a degumming agent, either phosphoric acid or citric acid, is added at a rate of 0.1-0.5% of the weight of the Ganoderma lucidum spore oil. The mixture is stirred and mixed for 30-60 minutes, and then centrifuged at 3000-5000 r / min for 10-20 minutes to separate the colloids and impurities. Subsequently, the centrifuged oil phase is filtered through a microporous membrane with a pore size of 0.2-0.5 μm and a filtration pressure of 0.1-0.3 MPa to remove residual particles and moisture, thus obtaining pretreated Ganoderma lucidum spore oil.

3. The method for purifying Ganoderma lucidum spore oil without plasticizers according to claim 2, characterized in that: The step of filtration through a microporous membrane specifically includes: First, a pre-filter with a pore size of 5-10 μm is used to coarsely filter the degummed oil phase at a filtration pressure of 0.05-0.1 MPa to remove larger particulate impurities. Then, a fine filter with a pore size of 0.2-0.5 μm is used for fine filtration at a filtration pressure of 0.1-0.3 MPa to ensure that the oil phase is clear.

4. The purification method for Ganoderma lucidum spore oil without plasticizers according to claim 1, characterized in that: The supercritical fluid extraction in step two uses an entrainer, which is at least one of ethanol and methanol, and the amount added is 5-15% of the weight of supercritical carbon dioxide. The entrainer is premixed with carbon dioxide before extraction. The mixing method is to inject the entrainer into the carbon dioxide stream at a flow rate of 1-3 mL / min using a high-pressure pump, and mix it in a static mixer at a pressure of 10-20 MPa for 5-10 minutes to form a homogeneous supercritical fluid system. The extraction process is carried out in an extraction vessel, and the loading of Ganoderma lucidum spore oil is 30-60% of the volume of the extraction vessel, and the carbon dioxide flow rate is 10-30 L / h.

5. The method for purifying Ganoderma lucidum spore oil without plasticizers according to claim 4, characterized in that: When the entrainer is ethanol, its concentration is above 95%, and the mixing ratio of ethanol and carbon dioxide is controlled by a mass flow meter. The amount of ethanol added is 8-12%. The extraction vessel is equipped with a temperature control system with a temperature control accuracy of ±1°C and a pressure control accuracy of ±0.5MPa. During the extraction process, samples are taken every 20-30 minutes to test the plasticizer content until the plasticizer residue is below 0.1mg / kg, at which point the extraction is terminated.

6. The purification method for Ganoderma lucidum spore oil without plasticizers according to claim 1, characterized in that: The supercritical carbon dioxide fluid in step two is obtained through a multi-stage extraction method, including a first-stage extraction and a second-stage extraction. The first-stage extraction pressure is 20-25 MPa and the temperature is 35-45°C, which removes most of the weakly polar plasticizers. The second-stage extraction pressure is 30-35 MPa and the temperature is 50-60°C, which removes the residual strongly polar plasticizers. The extraction time for each stage is 1-2 hours, and the carbon dioxide flow rate is 15-25 L / h. The two stages are switched by a valve to avoid cross-contamination. The multi-stage extraction system consists of extraction vessels connected in series, each with the same volume. The Ganoderma lucidum spore oil is processed in batches in each extraction vessel.

7. The method for purifying Ganoderma lucidum spore oil without plasticizers according to claim 1, characterized in that: The supercritical fluid extraction in step two is carried out under stirring conditions, with the stirring speed set to 100-300 r / min. During the extraction process, the density of the carbon dioxide fluid is adjusted in real time and controlled within the range of 0.5-0.8 g / mL. The extraction time is dynamically adjusted according to the amount of oil loaded, with an extraction time of 0.5-1 hour per kilogram of oil.

8. The method for purifying Ganoderma lucidum spore oil without plasticizers according to claim 1, characterized in that: The separation in step three is achieved through multi-stage decompression, including primary and secondary separation. Primary separation is carried out in a separation vessel, where the pressure is reduced from the extraction pressure to 10-15 MPa and the temperature is maintained at 35-45°C, causing most of the carbon dioxide to vaporize and carry away the plasticizer. Secondary separation is carried out in another separation vessel, where the pressure is further reduced to 5-8 MPa and the temperature is adjusted to 25-30°C, causing the remaining plasticizer to precipitate. The separated carbon dioxide is recovered through a condenser and recycled. During the separation process, an ultrasonic auxiliary device is installed in the separation vessel, with an ultrasonic frequency of 20-40 kHz, a power of 100-300 W, and an action time of 10-20 minutes.

9. The method for purifying Ganoderma lucidum spore oil without plasticizers according to claim 8, characterized in that: The analytical separation step in step three 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, and the amount added is 1-3% of the weight of Ganoderma lucidum spore oil. The adsorbent is pre-loaded into the filter screen of the separation vessel before analytical separation, and the contact time with the oil is 30-60 minutes. After adsorption, the adsorbent is separated by a filter press at a pressure of 0.2-0.5 MPa to ensure the oil is clear. The adsorbent can be regenerated and reused by calcining at 300-400°C for 1-2 hours.

10. The method for purifying Ganoderma lucidum spore oil without plasticizers according to claim 1, characterized in that: The post-processing in step four includes solvent removal and drying. Solvent removal is performed using a vacuum solvent removal device with a vacuum level of 0.05-0.1 MPa, a temperature of 50-60°C, and a solvent removal time of 1-2 hours to remove residual carbon dioxide and entrainers. Drying is performed using either spray drying or vacuum drying. During spray drying, the inlet temperature is 150-200°C, the outlet temperature is 80-100°C, and the atomization pressure is 0.3-0.6 MPa. During vacuum drying, the vacuum level is 0.01-0.05 MPa, the temperature is 40-50°C, and the drying time is 2-3 hours. After drying, the moisture content of the Ganoderma lucidum spore oil is less than 0.1%, and it is stored in nitrogen packaging.

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