A method for preparing organic Ganoderma lucidum spore powder under CO2-pure water dual-medium ultra-high pressure synergy

Abstract

The application provides a kind of C-pure water double medium ultra-high pressure coordinated organic broken wall ganoderma spore powder preparation method, belongs to the technical field of organic health food processing.The method comprises the following steps: (1) freeze-drying organic ganoderma spore powder after vacuum packaging, is treated with ultra-high pressure, obtains the organic ganoderma spore powder of first ultra-high pressure treatment;(2) the organic ganoderma spore powder of first ultra-high pressure treatment obtained in step (1) is freeze-dried again and placed in carbon dioxide medium, is treated with second ultra-high pressure, obtains the organic ganoderma spore powder of second ultra-high pressure treatment;(3) the organic ganoderma spore powder of second ultra-high pressure treatment obtained in step (2) is packaged, obtains organic broken wall ganoderma spore powder.Compared with the traditional liquid nitrogen ultra-low temperature broken wall method, the method can balance high breakage rate and active ingredient retention, and has the advantages of simple process, low energy consumption, easy to large-scale production.

Description

Technical Field

[0001] This invention relates to the field of organic health food processing technology, and more particularly to a C - A method for preparing organic Ganoderma lucidum spore powder by synergistic ultra-high pressure with pure water as the dual medium. Background Technology

[0002] Reishi spore powder has various health benefits, including boosting immunity, regulating blood lipids, providing antioxidant effects, protecting the liver, and improving sleep. Reishi spores are tiny, oval-shaped reproductive cells ejected from the cap of the reishi mushroom during its growth and maturation process. They possess an extremely tough cell wall structure. The spore wall is composed of chitin, cellulose, and silica, making it difficult for the human body to directly digest and absorb the active ingredients within the spores, such as reishi polysaccharides, triterpenoids, and proteins. Therefore, cell wall breaking is a crucial step in the processing of reishi spore powder.

[0003] Currently, commonly used methods for breaking down the cell walls of Ganoderma lucidum spore powder mainly include mechanical pulverization, low-temperature physical cell wall breaking, and wet temperature difference ultra-high pressure cell wall breaking. Among them, mechanical pulverization uses an ultra-micro pulverizer for cell wall breaking, but it easily generates high temperatures, which can damage active ingredients, and the cell wall breaking rate is limited; low-temperature physical cell wall breaking, such as liquid nitrogen freezing and crushing technology, can reduce the loss of heat-sensitive components, but it consumes a lot of energy and the cell wall breaking is uneven; wet temperature difference ultra-high pressure cell wall breaking requires multiple steps such as soaking, freezing, liquid nitrogen treatment, and microwave thawing to achieve cell wall breaking, which is a complex process and may introduce contamination during the process.

[0004] Furthermore, existing methods for sterilizing spore powder mostly employ irradiation or heat treatment, which can easily lead to the degradation of active ingredients and affect product quality. Ultra-high pressure (UHPP) technology, as a non-thermal processing technique, has shown potential in sterilization and cell wall disruption in the food and pharmaceutical industries. Studies have shown that UHPP extraction of polysaccharides from Ganoderma lucidum spore powder yields 37.1% higher than water extraction. However, UHPP alone cannot simultaneously achieve a high cell wall disruption rate and the retention of active ingredients, and its effect on disrupting the inner structure of the spores is limited.

[0005] Compared to conventional Ganoderma lucidum spore powder, organic Ganoderma lucidum spore powder must strictly comply with organic product certification standards (China GB / T 19630.1-2011 "Organic Products Part 1: Production", GB / T 19630.4-2011 "Organic Products Part 4: Management Systems", EU EC 834 / 2007 Regulation, and US NOP organic standards) throughout the entire process from Ganoderma lucidum cultivation and spore collection to deep processing. Among these, the rigid constraints directly related to processing include: Chemical intervention is strictly prohibited: The use of any chemical additives (such as preservatives and stabilizers), biological enzyme preparations (such as cellulase and pectinase), and organic solvents (such as ethanol and acetone) is strictly prohibited, and conventional enzymatic hydrolysis methods and chemically assisted cell wall breaking techniques are excluded. Physical processing is strictly limited: Irradiation sterilization is explicitly prohibited (GB / T 19630.4-2011 Clause 6.2.3 stipulates that "organic products shall not be irradiated"), and high-temperature processing is restricted (temperatures exceeding 60℃ may destroy the "natural activity" characteristics of organic products). Zero tolerance for pollution risks: Processing equipment and workshops must be certified organic, and the use of general equipment that may introduce impurities such as metals and microorganisms is prohibited (such as conventional mechanical crushers that are prone to producing metal wear particles). A full-process traceability system must also be established.

[0006] Current Ganoderma lucidum spore powder processing technologies cannot meet organic standards, and face the following technical bottlenecks: ① The contradiction between efficient cell wall breaking and organic purity: Conventional mechanical pulverization methods pose a risk of contamination, while technologies that meet purity requirements (such as low-temperature freeze drying) have insufficient cell wall breaking efficiency; ② The contradiction between thorough sterilization and no irradiation / no chemical intervention: Organic standards prohibit irradiation and chemical sterilization, while conventional physical sterilization (such as high temperature) will destroy active ingredients; ③ The contradiction between the preservation of active ingredients and processing efficiency: Mild processing (such as natural drying) preserves activity well but is inefficient, while efficient processing (such as high-temperature treatment) will lead to loss of activity.

[0007] Therefore, developing a method for preparing organic Ganoderma lucidum spore powder that fully complies with organic certification standards while also achieving high cell wall breakage rate, excellent sterilization effect, and high retention of active ingredients is a technical problem that urgently needs to be solved in this field, and it is also a core requirement for promoting the upgrading of the organic Ganoderma lucidum industry. Summary of the Invention

[0008] The purpose of this invention is to provide a method for preparing organic broken-cell Ganoderma lucidum spore powder that fully complies with organic certification standards, has a simple process, a cell wall breakage rate of ≥99.5%, can retain active ingredients to the maximum extent, and meets microbial indicators, as well as the resulting organic broken-cell Ganoderma lucidum spore powder product.

[0009] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a C -A method for preparing organic Ganoderma lucidum spore powder using a dual-medium ultra-high pressure system with pure water, comprising the following steps: (1) After freeze-drying the organic Ganoderma lucidum spore powder, vacuum-pack it and then subject it to ultra-high pressure treatment to obtain organic Ganoderma lucidum spore powder with initial ultra-high pressure treatment; (2) After the organic Ganoderma lucidum spore powder obtained in step (1) is freeze-dried again, it is placed under carbon dioxide medium and subjected to a second ultra-high pressure treatment to obtain organic Ganoderma lucidum spore powder subjected to a second ultra-high pressure treatment. (3) Package the organic Ganoderma lucidum spore powder obtained in step (2) after secondary ultra-high pressure treatment to obtain organic broken Ganoderma lucidum spore powder.

[0010] Preferably, the freeze-drying temperature in step (1) is -15 to -5°C, and the moisture content of the freeze-dried organic Ganoderma lucidum spore powder is 3 to 6%.

[0011] Preferably, the ultra-high pressure treatment in step (1) is carried out in pure water medium, the pressure of the ultra-high pressure treatment is 400~600MPa, and the pressure holding time is 5~10min.

[0012] Preferably, the temperature for the second freeze-drying in step (2) is -15 to -5°C, and the moisture content of the freeze-dried organic Ganoderma lucidum spore powder is 4 to 7%.

[0013] Preferably, the carbon dioxide medium in step (2) is organically certified food-grade liquid carbon dioxide; the purity of the carbon dioxide is ≥99.99%.

[0014] Preferably, the pressure during the second ultra-high pressure treatment in step (2) is 200~300MPa, and the pressure holding time is 2~8min.

[0015] Preferably, food-grade aluminum foil bags are used for packaging in step (3).

[0016] This invention provides an organic broken-cell wall Ganoderma lucidum spore powder prepared according to the method described above.

[0017] This invention provides an application of the aforementioned organic broken-cell wall Ganoderma lucidum spore powder in the preparation of products that enhance immunity, assist in lowering blood lipids, provide antioxidant effects, assist in protecting against chemically induced liver damage, or improve sleep.

[0018] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a method for preparing organic Ganoderma lucidum spore powder with cell wall disruption through controlled moisture content and a two-stage ultra-high pressure treatment. The first ultra-high pressure treatment is conducted under high pressure to induce microcracks in the spore wall. The second ultra-high pressure treatment is carried out in a carbon dioxide medium, utilizing the permeation of carbon dioxide molecules to further enhance the cell wall disruption effect and increase the dissolution of active ingredients, especially fat-soluble components (such as Ganoderma triterpenes), thereby improving bioavailability. The two ultra-high pressure treatments achieve a cell wall disruption rate of over 99.5% for the organic Ganoderma lucidum spore powder. Compared to traditional methods such as liquid nitrogen cryogenic cell wall disruption, this method achieves a balance between high cell wall disruption rate and retention of active ingredients, and has the advantages of simple process, low energy consumption, and ease of large-scale production.

[0019] The method of this invention fully complies with the requirements of organic food processing standards. It innovatively uses pure water and organically licensed CO2 as ultra-high pressure media to simultaneously achieve physical cell wall disruption and physical sterilization, thus achieving a balance between compliance and high efficiency in organic processing.

[0020] This invention is currently the only cell wall breaking and sterilization technology solution whose entire process, from raw material pretreatment to final packaging, is precisely designed to meet organic processing standards. Its closed-loop process of "pre-cracking-embrittlement-synergy" systematically solves the problems of pollution, violations, or substandard results that may occur in existing technologies at each stage.

[0021] This invention achieves, for the first time, highly efficient cell wall disruption, thorough sterilization, and preservation of active ingredients in organic Ganoderma lucidum spore powder, filling a gap in the organic Ganoderma lucidum spore powder processing industry. This invention is currently the only compliant and efficient technical solution applicable to the deep processing of organic Ganoderma lucidum spore powder, and the product can be widely used in the organic health food field. It also possesses the potential for technological platformization and has significant industrial value. Furthermore, the method of this invention can provide technical support for technicians processing other hard-shelled raw materials, such as pine pollen and cordyceps spore powder, showing broad application prospects. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0023] Figure 1 This is a schematic diagram of the overall structure of a supercritical carbon dioxide equipment for organic applications. 1-High pressure cylinder, 2-First four-way pipe, 3-Pressure relief valve, 4-Pressure sensor, 5-Stop valve, 6-Check valve, 7-Boost pump, 8-Media input pipe, 9-Second four-way pipe, 10-Safety valve, 11-Explosion-proof valve. Figure 2 This is a process flow diagram for preparing organic cell wall broken Ganoderma lucidum spore powder according to the present invention; Figure 3 The calculated results of the cell wall breakage rate of organic Ganoderma lucidum spore powder in different treatment groups; Figure 4 The results show the determination of the active ingredient content of organic broken-cell wall Ganoderma lucidum spore powder in different treatment groups; Figure 5 The structure of organic Ganoderma lucidum spore powder that has not undergone ultra-high pressure cell wall breaking treatment; Figure 6 The structure of organic Ganoderma lucidum spore powder after ultra-high pressure cell wall disruption treatment; Figure 7 , Figure 8 This is a test report on the crude polysaccharides, total triterpenes, lead, total mercury, total arsenic, and cell wall breakage rate of organic Ganoderma lucidum spore powder (i.e., organic broken cell wall Ganoderma lucidum spore powder); Figure 9 , Figure 10This is a test report on the polysaccharides, total triterpenes, and cell wall breakage rate of ultra-high pressure cell wall broken organic Ganoderma lucidum spore powder (i.e., organic cell wall broken Ganoderma lucidum spore powder). Detailed Implementation

[0024] This invention provides a C -A method for preparing organic Ganoderma lucidum spore powder using a dual-medium ultra-high pressure system with pure water, comprising the following steps: (1) After freeze-drying the organic Ganoderma lucidum spore powder, vacuum-pack it and then subject it to ultra-high pressure treatment to obtain organic Ganoderma lucidum spore powder with initial ultra-high pressure treatment; (2) After the organic Ganoderma lucidum spore powder obtained in step (1) is freeze-dried again, it is placed under carbon dioxide medium and subjected to a second ultra-high pressure treatment to obtain organic Ganoderma lucidum spore powder subjected to a second ultra-high pressure treatment. (3) Package the organic Ganoderma lucidum spore powder obtained in step (2) after secondary ultra-high pressure treatment to obtain organic broken Ganoderma lucidum spore powder.

[0025] In this invention, the freeze-drying temperature in step (1) is -15~-5℃, preferably -12~-8℃, and more preferably -10℃, and the moisture content of the freeze-dried organic Ganoderma lucidum spore powder is 3~6%, preferably 4~5%, and more preferably 4.5%.

[0026] Low-temperature freeze drying can remove free water from the spore wall, enhance the hydrogen bonding between cellulose and chitin molecules, and change the spore wall from tough to brittle, reducing the resistance to subsequent wall breaking. At the same time, low-temperature freeze drying can also inhibit the activity of polyphenol oxidase, peroxidase and other enzymes, avoiding the oxidative degradation of Ganoderma lucidum polysaccharides and triterpenes.

[0027] The moisture content of Ganoderma lucidum spore powder also has a significant impact on the subsequent cell wall breaking effect. If the moisture content is too high, the spore wall will be tough, resulting in a lower cell wall breaking rate. If the moisture content is too low, the spore powder will easily absorb moisture and clump together, and the risk of oxidation of active ingredients will increase.

[0028] In this invention, the ultra-high pressure treatment in step (1) is carried out in a pure water medium. The pressure during ultra-high pressure treatment is 400~600MPa, preferably 450~550MPa, more preferably 500MPa, and the holding time is 5~10min, preferably 6~9min, more preferably 7~8.5min, and more preferably 8min. The initial ultra-high pressure treatment uses pure water as the pressure transmission medium, causing uniform microcracks to appear in the tough spore wall.

[0029] In this invention, the temperature for the second freeze-drying in step (2) is -15 to -5°C, preferably -12 to -8°C, and even more preferably -10°C. The moisture content of the freeze-dried organic Ganoderma lucidum spore powder is 4 to 7%, preferably 5 to 6%, and even more preferably 5.5%.

[0030] In this invention, the carbon dioxide medium in step (2) is organically certified food-grade liquid carbon dioxide; the purity of the carbon dioxide is ≥99.99%.

[0031] In this invention, the pressure during the second ultra-high pressure treatment in step (2) is 200~300MPa, preferably 220~280MPa, more preferably 250MPa, and the pressure holding time is 2~8min, preferably 3~7min, more preferably 4~6min, and more preferably 5min.

[0032] The second ultra-high pressure treatment is carried out in a carbon dioxide medium. Carbon dioxide can rapidly penetrate into the spores through the microcracks created by the initial ultra-high pressure, forming a concentration gradient inside and outside the spore. During the subsequent depressurization process, the carbon dioxide inside the spore rapidly expands, generating enormous internal pressure. Combined with the squeezing action of the external high pressure, this causes the spore wall to completely rupture through the microcracks, achieving explosive cell disruption. Simultaneously, the acidification effect of carbon dioxide and the anaerobic environment work synergistically to effectively kill microorganisms, achieving sterilization. Carbon dioxide is the only medium that simultaneously satisfies the requirements of "organic licensing, physical cell disruption, and physical sterilization"—other gases (such as nitrogen and air) cannot achieve this penetration-explosive cell disruption and have poor sterilization effects.

[0033] In this invention, food-grade aluminum foil bags are used for packaging in step (3).

[0034] This invention provides an organic broken-cell wall Ganoderma lucidum spore powder prepared according to the method described above.

[0035] This invention provides an application of the aforementioned organic broken-cell wall Ganoderma lucidum spore powder in the preparation of products that enhance immunity, assist in lowering blood lipids, provide antioxidant effects, assist in protecting against chemically induced liver damage, or improve sleep.

[0036] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0037] The fresh organic Ganoderma lucidum spore powder used in the following examples was from the same batch and was organically certified. The fresh organic Ganoderma lucidum spore powder was sieved through a 100-mesh stainless steel sieve in an organic processing workshop certified by GB / T 19630.4-2011, and then transferred to a dedicated organic temporary storage tank for storage for ≤2 hours. The preparation of organic broken-cell wall Ganoderma lucidum spore powder was carried out entirely under organic processing conditions conforming to the GB / T 19630 series standards. All equipment (such as sieves, temporary storage tanks, ultra-high pressure equipment, freeze dryers, and supercritical carbon dioxide equipment) had to be certified for organic product processing, and each batch underwent cleaning verification according to organic cleaning procedures after processing to ensure that metal residues were <0.01 mg / kg and microbial residues were <10 CFU / g.

[0038] The organic-specific supercritical carbon dioxide equipment used in this invention has the application number 2025214320579 and is entitled "An Ultra-High Pressure Processing Equipment Based on a Gas Medium"; its overall structural schematic diagram is shown below. Figure 1 As shown, the processing flow is as follows: First, the material to be processed is added to the inner cavity of the high-pressure cylinder 1. Then, the plunger is installed into the high-pressure cylinder 1 to close it. The shut-off valve 5 and the check valve 6 are opened, and the booster pump 7 is turned on to fill the high-pressure cylinder 1 with gas medium and gradually increase the pressure to a certain level. The shut-off valve 5 is closed, and then pressure is applied to the plunger, causing the plunger to move inside the high-pressure cylinder 1 and gradually compress the space inside the high-pressure cylinder 1, thereby increasing the pressure inside the high-pressure cylinder 1 to a large value and maintaining the pressure for a period of time to achieve full processing of the material. During the pressurization process, the pressure sensor 4 can monitor the pressure inside the high-pressure cylinder 1 in real time, which facilitates precise pressure control. After processing is completed, the pressure relief valve 3 is opened to release the pressure, and then the high-pressure cylinder 1 is opened to remove the material from the high-pressure cylinder 1.

[0039] Example 1

[0040] A type of C -A method for preparing organic Ganoderma lucidum spore powder using a dual-medium ultra-high pressure system with pure water, the steps of which are as follows: (1) Transfer fresh organic Ganoderma lucidum spore powder into an organic freeze dryer and freeze dry it at -15℃ until the moisture content is 3%, and then vacuum pack it. Place the vacuum-packed organic Ganoderma lucidum spore powder in an ultra-high pressure device, inject food-grade pure water as the pressure medium, and hold it under pressure of 400MPa for 5 minutes. After depressurization, take it out to obtain organic Ganoderma lucidum spore powder that has undergone the first ultra-high pressure treatment. (2) The organic Ganoderma lucidum spore powder obtained in step (1) under the first ultra-high pressure treatment is transferred to an organic freeze dryer and freeze-dried again at -15℃ until the moisture content is 4%. Then it is transferred to an organic supercritical carbon dioxide equipment and organic certified food-grade liquid carbon dioxide (purity ≥99.99%) is introduced. The pressure is maintained at 200MPa for 2 minutes. After depressurization, the carbon dioxide is recovered (recovery rate ≥95%). The material is taken out in the organic clean area to obtain organic Ganoderma lucidum spore powder under the second ultra-high pressure treatment. (3) The organic Ganoderma lucidum spore powder obtained in step (2) after secondary ultra-high pressure treatment is packaged in a clean room environment to obtain the finished organic broken cell wall Ganoderma lucidum spore powder. The packaging bags used are organic certified food-grade aluminum foil packaging bags, which are vacuumed to -0.1MPa and heat-sealed using an organic special vacuum packaging machine.

[0041] Example 2

[0042] A type of C -A method for preparing organic Ganoderma lucidum spore powder using a dual-medium ultra-high pressure system with pure water, the steps of which are as follows: (1) Transfer fresh organic Ganoderma lucidum spore powder into an organic freeze dryer and freeze dry it at -5℃ until the moisture content is 6%, and then vacuum pack it. Place the vacuum-packed organic Ganoderma lucidum spore powder in an ultra-high pressure device, inject food-grade pure water as the pressure medium, and hold it under pressure of 600MPa for 10 minutes. After depressurization, take it out to obtain organic Ganoderma lucidum spore powder that has undergone the first ultra-high pressure treatment. (2) The organic Ganoderma lucidum spore powder obtained in step (1) under the first ultra-high pressure treatment is transferred to an organic freeze dryer and freeze-dried again at -5℃ until the moisture content is 7%. Then it is transferred to an organic supercritical carbon dioxide equipment and organic certified food-grade liquid carbon dioxide (purity ≥99.99%) is introduced. The pressure is maintained at 300MPa for 8 minutes. After depressurization, the carbon dioxide is recovered (recovery rate ≥95%). The material is taken out in the organic clean area to obtain organic Ganoderma lucidum spore powder under the second ultra-high pressure treatment. (3) The organic Ganoderma lucidum spore powder obtained in step (2) after secondary ultra-high pressure treatment is packaged in a clean room environment to obtain the finished organic broken cell wall Ganoderma lucidum spore powder. The packaging bags used are organic certified food-grade aluminum foil packaging bags, which are vacuumed to -0.1MPa and heat-sealed using an organic special vacuum packaging machine.

[0043] Example 3

[0044] A type of C -A method for preparing organic Ganoderma lucidum spore powder using a dual-medium ultra-high pressure system with pure water, the steps of which are as follows: (1) Transfer fresh organic Ganoderma lucidum spore powder into an organic freeze dryer and freeze dry it at -10℃ until the moisture content is 5%, and then vacuum pack it. Place the vacuum-packed organic Ganoderma lucidum spore powder in an ultra-high pressure device, inject food-grade pure water as the pressure medium, and hold it under pressure of 500MPa for 8 minutes. After depressurization, take it out to obtain organic Ganoderma lucidum spore powder that has undergone the first ultra-high pressure treatment. (2) The organic Ganoderma lucidum spore powder obtained in step (1) under the first ultra-high pressure treatment is transferred to an organic freeze dryer and freeze-dried again at -10℃ until the moisture content is 5.5%. Then it is transferred to an organic supercritical carbon dioxide equipment and organic certified food-grade liquid carbon dioxide (purity ≥99.99%) is introduced. The pressure is maintained at 250MPa for 5 minutes. After depressurization, the carbon dioxide is recovered (recovery rate ≥95%). The material is taken out in the organic clean area to obtain organic Ganoderma lucidum spore powder under the second ultra-high pressure treatment. (3) The organic Ganoderma lucidum spore powder obtained in step (2) after secondary ultra-high pressure treatment is packaged in a clean room environment to obtain the finished organic broken cell wall Ganoderma lucidum spore powder. The packaging bags used are organic certified food-grade aluminum foil packaging bags, which are vacuumed to -0.1MPa and heat-sealed using an organic special vacuum packaging machine.

[0045] Comparative Example 1

[0046] The difference between this comparative example and Example 3 is that step (2) is omitted. In this comparative example, the organic Ganoderma lucidum spore powder that has undergone initial ultra-high pressure treatment is packaged in a cleanroom environment to obtain the finished organic broken-cell wall Ganoderma lucidum spore powder. The remaining steps and parameters are the same as in Example 3.

[0047] Comparative Example 2

[0048] The difference between this comparative example and Example 3 is that in step (2), the freeze-dried organic Ganoderma lucidum spore powder is placed in an ultra-high pressure device, and food-grade pure water is injected as the pressure medium for a second ultra-high pressure treatment to obtain organic Ganoderma lucidum spore powder treated with a second ultra-high pressure. The remaining steps and parameters are the same as in Example 3.

[0049] Comparative Example 3

[0050] The difference between this comparative example and Example 3 is that the pressure during the ultra-high pressure treatment in step (1) is different. This comparative example undergoes ultra-high pressure treatment at a pressure of 200 MPa. The remaining steps and parameters are the same as in Example 3.

[0051] Comparative Example 4

[0052] The difference between this comparative example and Example 3 is that, in step (1), the fresh organic Ganoderma lucidum spore powder was not freeze-dried, but directly vacuum-packed and then subjected to ultra-high pressure treatment. The remaining steps and parameters are the same as in Example 3.

[0053] Experimental Example 1

[0054] 1. Cell wall breakage rate test

[0055] The cell wall breakage rates of the organic Ganoderma lucidum spore powders prepared in Examples 1-3 and Comparative Examples 1-4 were calculated according to the method disclosed by Gao Zhicheng et al. in "Determination of Cell Wall Breakage Rate of Ganoderma lucidum Spore Powder". Specifically, the unbroken organic Ganoderma lucidum spore powder and the organic broken organic Ganoderma lucidum spore powder from the same batch were dried separately in an oven at 60℃ for 5 hours. 0.1g of unbroken organic Ganoderma lucidum spore powder or 0.15g of broken organic Ganoderma lucidum spore powder were mixed with 5g of sucrose powder (ground three times and passed through a 100-mesh sieve), dissolved in distilled water, and 0.1mL of polysorbate-80 was added. The mixture was then brought to a final volume of 100mL in a volumetric flask and ultrasonically vibrated for 60 minutes to fully disperse the spores, yielding the spore suspension to be tested. The spore suspension to be tested was dropped into a hemocytometer (25 medium squares × 16 small squares), and observed and counted under a 400x optical microscope. The number of complete Ganoderma lucidum spores in the four corners and the five central squares of the hemocytometer was counted. Each sample was observed and counted three times, and the average value was taken.

[0056] Number of intact Ganoderma lucidum spores per gram of spore powder: N = n / 80 × 400 × 10,000 × V / m; where N is the number of intact Ganoderma lucidum spores per gram of spore powder (spores / g); n is the total number of intact Ganoderma lucidum spores in 80 small squares (spores); V is the volume of the spore dilution solution (mL); m is the mass of the sample (g); 400 is the number of small squares in the counting chamber of the hemocytometer; 10,000 is the volume of the counting chamber of the hemocytometer (0.1 mm²). 3 1 mL is equivalent to the volume of 10,000 hemocytometer chambers.

[0057] Cell wall breakage rate: X = (1 - N) B / N A ) × 100%; where X is the cell wall breakage rate of the broken organic Ganoderma lucidum spore powder, %; N B The number of intact Ganoderma lucidum spores per gram of broken-cell organic Ganoderma lucidum spore powder, expressed as spores / g; N A The number of complete Ganoderma lucidum spores per gram of organic Ganoderma lucidum spore powder, expressed as spores / g.

[0058] The results of the cell wall breakage rate calculation are shown in Table 1 and Figure 3 As shown, the cell wall breakage rate of the organic Ganoderma lucidum spore powder in Examples 1-3 was relatively high, reaching 99.5-99.8%, significantly higher than that in Comparative Examples 1-4. Comparative Example 1 underwent only one ultra-high pressure treatment, resulting in insufficient destruction of the spore wall; Comparative Example 2 did not undergo a second ultra-high pressure treatment in a carbon dioxide medium, indicating that the carbon dioxide medium plays an important role in sufficient cell wall breakage; the initial ultra-high pressure treatment pressure in Comparative Example 3 was too low, resulting in insufficient microcracks in the spore wall after initial cell wall breakage, and even with a subsequent second ultra-high pressure treatment in a carbon dioxide medium, the cell wall breakage effect was still unsatisfactory; Comparative Example 4 did not freeze-dry the fresh organic Ganoderma lucidum spore powder before the initial ultra-high pressure treatment, indicating that excessive moisture content is not conducive to spore wall destruction.

[0059] Table 1 Calculation results of cell wall breakage rate

[0060] 2. Determination of active ingredient content

[0061] The contents of crude polysaccharides, soluble polysaccharides, and total triterpenes in the organic Ganoderma lucidum spore powder prepared in undisturbed cell wall, as well as the organic Ganoderma lucidum spore powder prepared in Examples 1-3 and Comparative Examples 1-4, were determined. The polysaccharide content was determined using the phenol-sulfuric acid method, and the total triterpenes content was determined using the vanillin-glacial acetic acid colorimetric method. The results are shown in Table 2 and... Figure 4 As shown.

[0062] Table 2 Results of active ingredient content determination

[0063] It can be seen that, compared with the organic Ganoderma lucidum spore powder with intact cell wall, the organic Ganoderma lucidum spore powder with broken cell wall prepared in Examples 1-3 showed significantly increased contents of soluble polysaccharides and total triterpenes, indicating that the method of the present invention can significantly improve the dissolution of active ingredients. In contrast, the contents of polysaccharides and total triterpenes in the organic Ganoderma lucidum spore powder prepared by the methods of Comparative Examples 1-4 were significantly lower than those in Example 3, indicating that the two ultra-high pressure treatment methods and the step parameters used in the present invention have a significant impact on the dissolution of active ingredients in the organic Ganoderma lucidum spore powder.

[0064] 3. Microbiological index detection

[0065] Microbiological tests were performed on the organic Ganoderma lucidum spore powders prepared in Examples 1-3 and Comparative Examples 1-4, and the results are shown in Table 3. It can be seen that carbon dioxide medium treatment has a significant impact on the sterilization effect; using carbon dioxide as a medium during secondary ultra-high pressure treatment can significantly reduce the number of microorganisms.

[0066] Table 3 Results of Microbial Indicator Detection

[0067] 4. Microscopic observation

[0068] The structures of organic Ganoderma lucidum spore powder without ultra-high pressure cell disruption treatment and the organic Ganoderma lucidum spore powder prepared in Example 3 after ultra-high pressure cell disruption treatment were observed by electron microscopy. The results are as follows: Figure 5 and Figure 6 As shown in the figure, the organic Ganoderma lucidum spore powder that has not undergone ultra-high pressure cell disruption treatment has a complete structure. Figure 5 The structure was significantly damaged after ultra-high pressure cell disruption treatment. Figure 6 This demonstrates that the method of the present invention can improve the spore wall structure of organic Ganoderma lucidum spore powder and promote the release of active ingredients within the spores.

[0069] 5. Outsourced testing

[0070] The applicant previously commissioned PONY Testing Laboratory to test the content of active ingredients and the cell wall breakage rate of the organic Ganoderma lucidum spore powder prepared according to the method in Example 3. Two batches of samples were tested (the preparation method was the same as in Example 3, but the batches of organic Ganoderma lucidum spore powder and the content of active ingredients differed). The test results are as follows: Figures 7-8 as well as Figures 9-10 As shown in the figure, the organic Ganoderma lucidum spore powder prepared by the method of the present invention has a high cell wall breakage rate and a high content of active ingredients.

[0071] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing organic Ganoderma lucidum spore powder under CO2-pure water dual-medium ultra-high pressure synergy, characterized in that, Includes the following steps: (1) After freeze-drying the organic Ganoderma lucidum spore powder, vacuum-pack it and then subject it to ultra-high pressure treatment to obtain organic Ganoderma lucidum spore powder with initial ultra-high pressure treatment; (2) After the organic Ganoderma lucidum spore powder obtained in step (1) is freeze-dried again, it is placed under carbon dioxide medium and subjected to a second ultra-high pressure treatment to obtain organic Ganoderma lucidum spore powder subjected to a second ultra-high pressure treatment. (3) Package the organic Ganoderma lucidum spore powder obtained in step (2) after secondary ultra-high pressure treatment to obtain organic broken cell wall Ganoderma lucidum spore powder; The freeze-drying temperature in step (1) is -15 to -5°C, and the moisture content of the freeze-dried organic Ganoderma lucidum spore powder is 3 to 6%. The ultra-high pressure treatment described in step (1) is carried out in pure water medium, the pressure of the ultra-high pressure treatment is 400~600MPa, and the pressure holding time is 5~10min; The freeze-drying temperature in step (2) is -15 to -5°C, and the moisture content of the organic Ganoderma lucidum spore powder is 4% to 7% after freeze-drying. The carbon dioxide medium mentioned in step (2) is organically certified food-grade liquid carbon dioxide; the purity of the carbon dioxide is ≥99.99%; In step (2), the pressure during the second ultra-high pressure treatment is 200~300MPa, and the pressure holding time is 2~8min.

2. The method according to claim 1, characterized in that, In step (3), food-grade aluminum foil bags are used for packaging.

3. An organic broken-cell wall Ganoderma lucidum spore powder prepared according to any one of claims 1 to 2.

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