Ganoderma sinensis flash-soluble tablet and preparation method thereof

Abstract

The invention discloses a Ganoderma sinense flash-soluble tablet and a preparation method thereof, and belongs to the technical field of food health care, Ganoderma sinense sporocarp is subjected to deproteinization treatment to prevent precipitation generated by combination of tannic acid and Ganoderma sinense protein, and polysaccharide and triterpenes in Ganoderma sinense sporocarp are effectively retained and enriched after Ganoderma sinense sporocarp superfine powder is subjected to deproteinization treatment. In the preparation process, tannic acid serves as a structure trigger and is self-assembled with ganoderma sinense polysaccharide in deproteinized ganoderma sinense powder through hydrogen bonds to form a pH responsive hydration film, phenolic hydroxyl groups of tannic acid and alcoholic hydroxyl groups of beta-glucan form a multiple hydrogen bond network, aromatic rings are embedded into hydrophobic cavities of beta-glucan, and the compound is stabilized through the hydrophobic effect. By adopting hydroxypropyl-beta-cyclodextrin inclusion, the tannic acid is prevented from being combined with the ganoderma sinense sporocarp protein to form a precipitate in space. The Ganoderma sinensis flash-soluble tablet prepared by the invention is rich in triterpenes and polysaccharide components, can significantly improve immunity and regulate intestinal flora, and is suitable for health food development.

Description

Technical Field

[0001] This invention relates to the field of food and health care technology, specifically to a Ganoderma lucidum flash-dissolving tablet and its preparation method. Background Technology

[0002] Reishi mushroom, a precious treasure in the Chinese medicine treasury, is known as the "immortal herb" and is a good medicine for "strengthening the body's resistance and consolidating its foundation." It is usually divided into purple reishi (Ganoderma sinense) and red reishi (Ganoderma lucidum). The Shennong's Classic of Materia Medica lists purple reishi as a "superior" medicine, emphasizing its effects of "calming the mind and replenishing qi." Compared with red reishi, purple reishi has a higher content of unique ganoderic acid and euphorbia triterpenes. Its anti-inflammatory and hepatoprotective activities are significantly better than the common triterpenes in red reishi. Under the same cultivation conditions, purple reishi has a 15% to 20% lower adsorption rate of soil heavy metals (such as lead and cadmium) than red reishi, making it more suitable for safety-oriented product development.

[0003] Currently, most health products on the market using Ganoderma lucidum as a raw material employ spore powder. While spore powder, derived from the reproductive cells of Ganoderma lucidum, has a high concentration of active ingredients, its application is significantly limited: First, the outer layer of spore powder is encapsulated by chitin, requiring cell wall disruption to promote the dissolution of active ingredients. This not only greatly increases the complexity of the process and production costs, but also makes the unsaturated fatty acids prone to oxidation and rancidity after cell wall disruption, making it difficult to preserve in summer and necessitating the addition of antioxidants (such as vitamin E). Second, spore powder is mainly composed of fatty acids and a small amount of triterpenes, with relatively simple functional components, lacking the synergistic components unique to the fruiting body, such as sterols and alkaloids, thus limiting its overall health benefits. Third, spore powder is slightly bitter and has an earthy taste, resulting in poor palatability, and its low yield leads to high prices, making it difficult to meet the needs of developing mass-market health foods.

[0004] In contrast, the fruiting body of Ganoderma lucidum, as the mature form of Ganoderma lucidum, is rich in polysaccharides, triterpenes, sterols, and other synergistic functional components, offering advantages such as comprehensive composition, low cost, and good palatability. Furthermore, after ultrafine pulverization (D90≤45μm), the fibrous skeleton of the fruiting body forms a naturally porous structure, with a water permeation rate 40% higher than that of spore powder, making it more suitable for the development of instant formulations. However, the application of fruiting bodies in health products faces the following technical bottlenecks: the fruiting body contains approximately 7%–12% crude protein, among which basic proteins (such as lectins) readily combine with polyphenols (such as tannins) to form irreversible precipitation, affecting not only the product's appearance and taste but also leading to the loss of active ingredients and a decline in function. This severely limits the development and application of fruiting bodies in high-quality instant products. Therefore, overcoming the protein-polyphenol precipitation problem in the fruiting body, maximizing the retention of its active ingredients, and fully utilizing the raw material value of the fruiting body has become a pressing technical challenge in the development of Ganoderma lucidum health foods. Summary of the Invention

[0005] This invention provides a Ganoderma lucidum flash-dissolving tablet and its preparation method, which effectively solves the technical problem of precipitation of fruiting body protein-polyphenols when preparing health products using fruiting bodies as raw materials, resulting in significant loss of active ingredients, decreased function, and poor appearance and taste of health products. At the same time, it provides a Ganoderma lucidum flash-dissolving tablet rich in triterpenes and polysaccharides, which can significantly improve immunity and regulate intestinal flora.

[0006] The first objective of this invention is to provide a method for preparing Ganoderma lucidum flash-dissolving tablets, comprising the following steps: Using ultrafine powder of Ganoderma lucidum fruiting bodies as raw material, the protein is removed to obtain a deproteinized fruiting body concentrate. The deproteinized fruiting body concentrate is then mixed with microcrystalline cellulose and spray-dried to obtain deproteinized Ganoderma lucidum powder.

[0007] The deproteinized Ganoderma lucidum powder is mixed with excipients to obtain a mixed powder. A solution of an ethanol inclusion complex of tannic acid and hydroxypropyl-β-cyclodextrin is atomized and sprayed into the mixed powder to obtain a soft material.

[0008] Using the soft material as a matrix, Ganoderma lucidum flash-dissolved tablets were prepared.

[0009] As a preferred embodiment, the preparation method of the deproteinized fruiting body concentrate is as follows: add an acetate buffer solution with a pH of 3.5 to 4.5 to the ultrafine powder of Ganoderma lucidum fruiting bodies, stir and centrifuge at room temperature, take the supernatant, adjust the pH to neutral, centrifuge again, retain the supernatant and adjust the pH to 5 to 6, concentrate under reduced pressure to obtain a deproteinized fruiting body concentrate with a solid content of 40% to 65%.

[0010] In a preferred embodiment, the mass ratio of the deproteinized fruiting body concentrate to microcrystalline cellulose is 1:0.2~0.4.

[0011] In a preferred embodiment, the concentrate and microcrystalline cellulose are dried by spray drying, wherein the inlet air temperature is controlled at 55℃~65℃ and the outlet air temperature is controlled at 40℃~45℃.

[0012] In a preferred embodiment, the protein residue of the deproteinized Ganoderma lucidum powder is 0.5% to 0.8%, and the polysaccharide retention rate is 95% to 98%.

[0013] In a preferred embodiment, the amount of tannic acid added is 0.8% to 1.2% by mass of the mixed powder, and the molar ratio of hydroxypropyl-β-cyclodextrin to tannic acid is 1.5 to 3:1.

[0014] In a preferred embodiment, the ethanol concentration in the ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is 60%~65%, and the mass-to-volume ratio of the mixed powder to the ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is 1g:0.25mL~0.35mL.

[0015] In a preferred embodiment, when the ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is atomized and sprayed into the mixed powder, the temperature of the mixed powder is controlled at 18°C ​​to 30°C and the moisture content is 8% to 15%.

[0016] In a preferred embodiment, the soft material is granulated, sized, and tableted to obtain Ganoderma lucidum flash-dissolving tablets. During granulation, an aqueous solution of ferric ammonium citrate with a mass concentration of 25% to 30% at 1°C to 4°C is sprayed onto the initial granules obtained after granulation. The mixture is then flash-evaporated at 65°C to 75°C for 2 to 4 minutes and finally dried at 50°C to 60°C until the moisture content is 2.0% to 3.0%, thus obtaining final granules. During tableting, magnesium stearate with a mass ratio of 0.5% to 1% is added as a lubricant based on the mass of the final granules and mixed with the final granules. The mixture is then allowed to stand and tableted.

[0017] In a preferred embodiment, the excipients include a co-disintegrant, a filler, and a surfactant; the co-disintegrant is composed of croscarmellose sodium, croscarmellose, and microcrystalline cellulose; the amount of each excipient added is calculated based on the total mass of the mixed powder, wherein the amount of croscarmellose sodium added is 2%~5%; the amount of croscarmellose added is 3%~8%; the amount of microcrystalline cellulose added is 2%~6%; the amount of surfactant polysorbate 80 added is 0.1%~0.5%; and mannitol is used as a filler to make up to 100%.

[0018] The second objective of this invention is to provide a Ganoderma lucidum flash-dissolving tablet prepared by the above-described preparation method.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention provides a method for preparing Ganoderma lucidum flash-dissolving tablets. Using ultrafine powder of Ganoderma lucidum fruiting bodies as raw material, the protein is removed to obtain a deproteinized fruiting body concentrate. This concentrate is then mixed with microcrystalline cellulose and spray-dried to obtain deproteinized Ganoderma lucidum powder. The deproteinized powder is mixed with excipients to obtain a mixed powder. An ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is atomized and sprayed into the mixed powder to obtain a soft material. Using this soft material as a matrix, Ganoderma lucidum flash-dissolving tablets are prepared. Since Ganoderma lucidum fruiting bodies contain approximately 7%–12% crude protein, and basic proteins (such as lectins) readily form irreversible precipitation with tannic acid, deproteinization of the fruiting bodies prevents tannic acid from binding with Ganoderma lucidum protein to form precipitation. Simultaneously, after deproteinization, the polysaccharides and triterpenes in the ultrafine powder of Ganoderma lucidum fruiting bodies are effectively retained and enriched. In the preparation process, this invention uses tannic acid as a structural trigger, which self-assembles with Ganoderma lucidum polysaccharides in deproteinized Ganoderma lucidum powder through hydrogen bonds to form a pH-responsive hydration membrane. The phenolic hydroxyl groups of tannic acid and the alcoholic hydroxyl groups of β-glucan, a cell wall derivative of deproteinized Ganoderma lucidum fruiting bodies, form a multiple hydrogen bond network. The aromatic ring of tannic acid is embedded in the hydrophobic cavity of β-glucan, stabilizing the complex (a supramolecular structure formed by the self-assembly of tannic acid molecules and Ganoderma lucidum polysaccharide molecules through hydrogen bonds and hydrophobic interactions) through hydrophobic interactions. Hydroxypropyl-β-cyclodextrin inclusion is used to spatially prevent tannic acid from binding with Ganoderma lucidum fruiting body proteins to form a precipitate.

[0020] After deproteinization, this invention mixes the deproteinized fruiting body concentrate with microcrystalline cellulose before spray drying. Because the deproteinized fruiting body concentrate is rich in polysaccharides and other viscous components, direct spray drying suffers from problems such as wall adhesion, nozzle clogging, and low efficiency. This invention addresses this by adding microcrystalline cellulose as a drying aid and carrier, playing a dual crucial role: First, as a carrier for spray drying, its porous structure and good adsorption properties effectively prevent the high-sugar concentrate from sticking to the walls and charring during the drying process, ensuring smooth drying and high retention of active ingredients. Second, as a component of the final tablet co-disintegrant, its early introduction allows for more uniform distribution in the overall formulation, thus creating a synergistic effect with subsequently added disintegrants and further optimizing the tablet's pore structure and disintegration performance.

[0021] The Ganoderma lucidum flash-dissolving tablets prepared by this invention are rich in triterpenes and polysaccharides, which can significantly improve immunity and regulate intestinal flora, making them suitable for health food development. This invention provides a method for preparing Ganoderma lucidum flash-dissolving tablets that retains as many of the effective components of Ganoderma lucidum as possible and fully utilizes the value of the Ganoderma lucidum fruiting body.

[0022] The tannic acid used in this invention is rich in phenolic hydroxyl groups, and the ferric ammonium citrate contains Fe. 3+The Ganoderma lucidum flash dissolution tablets of the present invention are prepared through a step-by-step isolation process of "embedded tannic acid skeleton + external iron ammonium delivery layer". Tannic acid is sprayed in the form of ethanol solution during the soft material preparation stage, and iron ammonium citrate is added in the form of cold aqueous solution during granulation to avoid the possible formation of blue-black complex precipitates from direct mixing. Attached Figure Description

[0023] Figure 1 This is a standard curve of the polysaccharide solution of the present invention.

[0024] Figure 2 This is the curve of the ursolic acid standard solution of the present invention. Detailed Implementation

[0025] To enable those skilled in the art to better understand and implement the technical solutions of this invention, the invention is further described below with reference to specific embodiments. However, the embodiments are not intended to limit the invention. Unless otherwise specified, the following test methods and detection methods are conventional methods; unless otherwise specified, the reagents and raw materials are commercially available.

[0026] Regarding the issues mentioned in the background of this invention: First, when preparing Ganoderma lucidum health products using spore powder as a raw material, the outer layer of the spore powder is encapsulated by chitin, which usually requires cell wall breaking to promote the dissolution of active ingredients. This not only significantly increases the complexity of the process and production costs, but also makes the unsaturated fatty acids prone to oxidation and rancidity after cell wall breaking, making it difficult to preserve in summer and requiring the addition of antioxidants, such as vitamin E. Spore powder is mainly composed of fatty acids and a small amount of triterpenes, with relatively simple functional components, lacking synergistic components such as sterols and alkaloids unique to the fruiting body, thus limiting its overall health benefits. Spore powder is slightly bitter and has an earthy taste, resulting in poor palatability, and its low yield leads to high prices, making it difficult to meet the needs of developing mass-market health foods. Second, when preparing health products using Ganoderma lucidum fruiting bodies as raw materials, the fruiting bodies contain approximately 7% to 12% crude protein, among which basic proteins (such as lectins) readily combine with polyphenolic substances, such as tannic acid, to form irreversible precipitation, which not only affects the appearance and taste of the product, but also leads to the loss of active ingredients and a decline in function. This seriously limits the development and application of fruiting bodies in high-quality instant products. To address the aforementioned technical issues, this invention provides a Ganoderma lucidum flash-dissolving tablet and its preparation method.

[0027] The technical solution of the present invention will be described in detail below.

[0028] This invention provides a method for preparing Ganoderma lucidum flash-dissolving tablets, comprising the following steps: Using ultrafine powder of Ganoderma lucidum fruiting bodies as raw material, the protein is removed to obtain a deproteinized fruiting body concentrate. The deproteinized fruiting body concentrate is mixed with microcrystalline cellulose and then spray-dried to obtain deproteinized Ganoderma lucidum powder.

[0029] The deproteinized Ganoderma lucidum powder is mixed with excipients to obtain a mixed powder. A solution of an ethanol inclusion complex of tannic acid and hydroxypropyl-β-cyclodextrin is atomized and sprayed into the mixed powder to obtain a soft material.

[0030] Using the soft material as a matrix, Ganoderma lucidum flash-dissolved tablets were prepared.

[0031] As a preferred embodiment, the preparation method of the deproteinized fruiting body concentrate is as follows: add an acetate buffer solution with a pH of 3.5 to 4.5 to the ultrafine powder of Ganoderma lucidum fruiting bodies, stir and centrifuge at room temperature, take the supernatant, adjust the pH to neutral, centrifuge again, retain the supernatant and adjust the pH to 5 to 6, concentrate under reduced pressure to obtain a deproteinized fruiting body concentrate with a solid content of 40% to 65%.

[0032] Basic proteins, such as lectins, which readily combine with polyphenols to form precipitates, typically have isoelectric points (pI) above 7.5. Based on the characteristic that proteins have the lowest solubility at pH equal to their isoelectric point, this invention utilizes an acidic buffer solution to effectively precipitate these basic proteins. When the pH is too low (<3.5), some acidic polysaccharides or triterpenic acids undergo hydrolysis or structural changes; when the pH is too high (>4.5), the precipitation efficiency of basic proteins decreases significantly. Therefore, this invention selects an acidic buffer solution with a pH of 3.5–4.5, which can efficiently precipitate basic proteins while maximally protecting the active ingredients in Ganoderma lucidum.

[0033] In the above technical solution, the endpoint of the vacuum concentration is determined by the solid content. When the solid content is below 40%, there is too much moisture, which is not conducive to spray drying after subsequent mixing with microcrystalline cellulose, resulting in high energy consumption and easy loss of active ingredients due to excessive heating time. When the solid content is above 65%, the concentrate is too viscous and has poor fluidity, making it difficult to mix evenly with microcrystalline cellulose and easily clogging the atomizing nozzle, leading to production interruption. Therefore, this invention limits the vacuum concentration to obtaining deproteinized fruiting bodies with a solid content of 45% to 60%. Within this range, the concentrate has a suitable viscosity, which can ensure the uniformity of mixing with excipients and achieve efficient and stable spray drying.

[0034] The deproteinized fruiting body concentrate, due to its high content of polysaccharides and other viscous components, suffers from problems such as adhesion to the walls, nozzle clogging, and low efficiency when directly spray-dried. This invention addresses this by adding microcrystalline cellulose as a drying aid and carrier. Utilizing its porous structure and oil- and water-absorbing properties, it effectively improves the material characteristics of the concentrate, making it suitable for spray drying. The amount of microcrystalline cellulose used must be matched to the solid content of the concentrate: when the solid content is high, it tends towards the upper limit of the dosage range; when the solid content is low, it tends towards the lower limit of the dosage range.

[0035] In a preferred embodiment, the mass ratio of the deproteinized fruiting body concentrate to microcrystalline cellulose is 1:0.2~0.4, preferably 1:0.3. This ratio depends on the solids content and viscosity of the deproteinized fruiting body concentrate. When the mass ratio of microcrystalline cellulose is lower than the specified 0.2, the mixture viscosity is too high, leading to severe wall adhesion during spray drying, low production efficiency, and easy degradation of the heat-sensitive active ingredient due to prolonged heating. When the mass ratio of microcrystalline cellulose is higher than the specified 0.4, the active ingredient is excessively diluted, resulting in a low content of active ingredients in the final product. This necessitates increasing the tablet size to achieve the desired dosage, reducing economic efficiency and product applicability. Within the above-defined range, the effective ingredient content and economic efficiency of the product can be maximized while ensuring smooth spray drying.

[0036] This invention involves mixing the deproteinized fruiting body concentrate with microcrystalline cellulose and then spray-drying it. The inlet air temperature is controlled at 55℃~65℃, and the outlet air temperature is controlled at 40℃~45℃. When the inlet air temperature is below 55℃, the moisture evaporation efficiency is too low, the drying is incomplete, the resulting powder has a high moisture content, is prone to clumping, and energy consumption increases significantly. When the inlet air temperature is above 65℃, although the drying efficiency can be improved, it will cause the degradation and charring of heat-sensitive active ingredients, such as polysaccharides and triterpenes, resulting in a decrease in product functionality. The outlet air temperature is a key indicator of the actual heating temperature of the material. Controlling the outlet air temperature to not exceed 45℃ ensures that the material is always in a mild environment, maximizing the protection of the bioactivity of Ganoderma lucidum active ingredients. At the same time, an outlet air temperature not lower than 40℃ is a necessary condition to ensure that the humidity inside the drying tower is low enough, to prevent the powder from deliquescing and adhering to the tower wall, and to ensure continuous and stable production operation.

[0037] It should be noted that the protein residue of the deproteinized Ganoderma lucidum powder prepared by this invention is 0.5%~0.8%, and the polysaccharide retention rate is 95%~98%.

[0038] In a preferred embodiment, the amount of tannic acid added is 0.8% to 1.2% by weight of the mixed powder, and the molar ratio of hydroxypropyl-β-cyclodextrin to tannic acid is 1.5 to 3:1. When the molar amount of hydroxypropyl-β-cyclodextrin is less than 1.5 (as defined herein), i.e., the amount of hydroxypropyl-β-cyclodextrin is insufficient, it cannot effectively encapsulate the tannic acid, and there is still a risk of binding with proteins. When the molar amount of hydroxypropyl-β-cyclodextrin is greater than 3 (as defined herein), the encapsulation effect is not significantly improved, but it will significantly increase the raw material cost and affect the subsequent granulation process. Therefore, the present invention limits the molar ratio of hydroxypropyl-β-cyclodextrin to tannic acid to 1.5 to 3:1, more preferably 1.8 to 2.2:1, within which the optimal balance between ensuring the encapsulation effect and economy can be achieved.

[0039] To prepare a soft material with superior performance, the ethanol concentration in the tannic acid and hydroxypropyl-β-cyclodextrin ethanol inclusion complex solution is 60%~65%, and the mass-to-volume ratio of the mixed powder to the tannic acid and hydroxypropyl-β-cyclodextrin ethanol inclusion complex solution is 1g:0.25mL~0.35mL. Within this range, it ensures that the powder is sufficiently and uniformly wetted to form a soft material suitable for granulation, while preventing problems such as sticking to the sieve and difficulty in drying due to excessive wetness. If the amount of the hydroxypropyl-β-cyclodextrin ethanol inclusion complex solution is less than the specified 0.25mL, insufficient wetting will result in the inability to form the soft material; if the amount of the hydroxypropyl-β-cyclodextrin ethanol inclusion complex solution is greater than 0.35mL, the soft material will be too sticky, which is not conducive to subsequent processes. More preferably, the mass-to-volume ratio is 1g:0.28mL~0.32mL, which is the optimal range for forming an ideal soft material.

[0040] To ensure the uniformity and stability of the Ganoderma lucidum flash-dissolving tablets, the temperature of the mixed powder is controlled at 18℃~30℃ and the moisture content at 8%~15% when the ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is atomized and sprayed into the mixed powder. Temperature control is crucial during the spray-forming process. Below 18℃, the ethanol evaporation rate is too slow, potentially leading to localized over-wetting and clumping; simultaneously, insufficient molecular thermal motion hinders the efficient hydrogen bond self-assembly reaction between tannic acid and polysaccharide chains. Above 30℃, ethanol evaporation is too rapid, causing the inclusion complex solution to dry prematurely on the powder surface, resulting in a false-drying phenomenon, uneven wetting, and failure to form a uniformly quality soft material; furthermore, excessively high temperatures can cause degradation of heat-sensitive active ingredients. Therefore, controlling the material temperature within 18℃~30℃ is critical. Within this range, both wetting and molecular bonding efficiency are guaranteed, ensuring product quality uniformity and stability. The moisture content of the mixed powder after spraying is a key indicator determining the quality of the soft material. When the moisture content is below 8%, the powder is insufficiently wetted, resulting in few and weak liquid bridges that cannot form effective agglomerates. The resulting soft material is loose and cannot be processed into qualified granules, leading to an extremely high powder yield. When the moisture content is above 15%, the powder is over-wetted, the soft material is too sticky, and it easily adheres and clumps on the granulator screen, resulting in decreased production efficiency and excessive drying load, leading to increased energy consumption. Therefore, the moisture content after spraying should be precisely controlled between 8% and 15%. Under these conditions, the tannic acid-cyclodextrin inclusion complex exhibits the best dispersibility and stability, the highest hydrogen bond self-assembly efficiency, and the resulting soft material has good plasticity and moderate viscosity, which is most conducive to the subsequent production of granules with uniform particle size and good flowability.

[0041] It should be noted that the Ganoderma lucidum flash-dissolving tablets of the present invention are obtained by granulation, sizing and tableting of the soft material. During sizing, an aqueous solution of ferric ammonium citrate with a mass concentration of 25% to 30% at 1℃ to 4℃ is sprayed onto the initial granules obtained after granulation. The mixture is then flash-evaporated at 65℃ to 75℃ for 2 to 4 minutes and finally dried at 50℃ to 60℃ until the moisture content is 2.0% to 3.0% to obtain the final granules. During tableting, magnesium stearate with a mass ratio of 0.5% to 1% is added as a lubricant based on the mass of the final granules and mixed with the final granules. The mixture is then allowed to stand and tableted.

[0042] To obtain Ganoderma lucidum flash-dissolving tablets with better performance, this invention also adds excipients to the deproteinized Ganoderma lucidum powder. The excipients include a co-disintegrant, a filler, and a surfactant. The co-disintegrant is composed of croscarmellose sodium, croscarmellose povidone, and microcrystalline cellulose. The amount of each excipient added is calculated based on the total mass of the mixed powder. The amount of croscarmellose sodium added is 2%~5%; the amount of croscarmellose added is 3%~8%; the amount of microcrystalline cellulose added is 2%~6%; the amount of surfactant polysorbate 80 added is 0.1%~0.5%; and mannitol is used as a filler to make up to 100%.

[0043] The invention will now be described in detail through the following embodiments and comparative examples.

[0044] Example 1 A method for preparing Ganoderma lucidum flash-dissolving tablets includes the following steps: S1, Raw material pretreatment: Add acetate buffer (pH=4.0) to the ultrafine powder of Ganoderma lucidum fruiting bodies at a material-to-liquid ratio of 1g:15mL. Stir at 25℃ for 30min, centrifuge at 8000rpm for 10min, discard the precipitate, adjust the pH of the supernatant to 7.0, centrifuge again to discard the precipitate, retain the supernatant, adjust the pH to 5.5, and concentrate under reduced pressure until the solid content is 45%, obtaining a deproteinized fruiting body concentrate. Mix the deproteinized fruiting body concentrate with microcrystalline cellulose at a mass ratio of 1:0.3, and spray dry with air inlet at 60℃ and air outlet at 42℃ to obtain deproteinized Ganoderma lucidum powder with a protein residue of 0.7% and a polysaccharide retention rate of 96.5%.

[0045] S2, weigh 400g of deproteinized Ganoderma lucidum powder, 35g of croscarmellose sodium, 55g of croscarmellose, 40g of microcrystalline cellulose, 467g of mannitol and 3g of polysorbate 80, put them into a V-type mixer and mix at 20 rpm for 15 minutes to obtain a mixed powder.

[0046] S3: 10.0g of tannic acid and 20.9g of hydroxypropyl-β-cyclodextrin inclusion complex were dissolved in 300mL of 65% ethanol to prepare an inclusion complex solution; 1000g of the mixed powder from S2 was placed in a fluidized bed, and the above inclusion complex solution was uniformly sprayed in at an atomization pressure of 0.3MPa, while controlling the material temperature at 30℃ throughout. After the spraying was completed, the moisture content of the material was measured to be 12%, resulting in a soft material.

[0047] S4. The soft material is fed into the hopper of a granulator (1.0 mm / 18 mesh), extruded and sieved at 100 rpm, and then placed in a fluidized bed dryer. It is initially dried at 55°C to a moisture content of 8.0%, yielding approximately 1010 g of initial granules. 30.3 g of ferric ammonium citrate is weighed and dissolved in 70 mL of 4°C pure water. The solution is stirred until completely dissolved to prepare a 30% (w / w) ferric ammonium citrate solution, which is maintained at 4°C. The initial granules are placed in a fluidized bed, and the prepared ferric ammonium citrate solution is sprayed in at an atomizing pressure of 0.15 MPa. The solution is then flash-evaporated at 70°C for 3 min, and finally dried at 60°C to a moisture content of 2.5%, yielding final granules. 10.1 g of magnesium stearate (1.0% of the final granule mass) is weighed as a lubricant and mixed with the final granules in a mixing drum. After standing in a sealed container for 30 min, the mixture is compressed into tablets to obtain Ganoderma lucidum flash-dissolving tablets.

[0048] Example 2 A method for preparing Ganoderma lucidum flash-dissolving tablets includes the following steps: S1, Raw material pretreatment: Add acetate buffer (pH=3.5) to the ultrafine powder of Ganoderma lucidum fruiting bodies at a material-to-liquid ratio of 1g:15mL. Stir at 25℃ for 30min, centrifuge at 8000rpm for 10min, discard the precipitate, adjust the pH of the supernatant to 7.0, centrifuge again to discard the precipitate, retain the supernatant, adjust the pH to 5.0, and concentrate under reduced pressure until the solid content is 40%, obtaining a deproteinized fruiting body concentrate. Mix the deproteinized fruiting body concentrate with microcrystalline cellulose at a mass ratio of 1:0.3, and spray dry with air inlet at 60℃ and air outlet at 42℃ to obtain deproteinized Ganoderma lucidum powder with a protein residue of 0.8% and a polysaccharide retention rate of 95%.

[0049] S2, weigh 400g of deproteinized Ganoderma lucidum powder, 35g of croscarmellose sodium, 55g of croscarmellose, 40g of microcrystalline cellulose, 467g of mannitol and 3g of polysorbate 80, put them into a V-type mixer and mix at 20 rpm for 15 minutes to obtain a mixed powder.

[0050] S3: Dissolve 8.0g of tannic acid and 17.9g of hydroxypropyl-β-cyclodextrin inclusion complex in 300mL of 60% ethanol to prepare inclusion complex solution; Place 1000g of the mixed powder from S2 in a fluidized bed and spray the above inclusion complex solution evenly with an atomization pressure of 0.3MPa, controlling the material temperature at 18℃ throughout. After spraying, the moisture content of the material is measured to be 8%, resulting in a soft material.

[0051] S4. The soft material is fed into the hopper of a granulator (1.0 mm / 18 mesh), extruded and sieved at 100 rpm, and then placed in a fluidized bed dryer. It is initially dried at 55°C to a moisture content of 8.0%, yielding approximately 1010 g of initial granules. 30.3 g of ferric ammonium citrate is weighed and dissolved in 70 mL of 4°C pure water. The solution is stirred until completely dissolved to prepare a 30% (w / w) ferric ammonium citrate solution, which is maintained at 4°C. The initial granules are placed in a fluidized bed, and the prepared ferric ammonium citrate solution is sprayed in at an atomizing pressure of 0.15 MPa. The solution is then flash-evaporated at 70°C for 3 min, and finally dried at 60°C to a moisture content of 2.5%, yielding final granules. 10.1 g of magnesium stearate (1.0% of the final granule mass) is weighed as a lubricant and mixed with the final granules in a mixing drum. After standing in a sealed container for 30 min, the mixture is compressed into tablets to obtain Ganoderma lucidum flash-dissolving tablets.

[0052] Example 3 A method for preparing Ganoderma lucidum flash-dissolving tablets includes the following steps: S1, Raw material pretreatment: Add acetate buffer (pH=4.2) to the ultrafine powder of Ganoderma lucidum fruiting bodies at a material-to-liquid ratio of 1g:15mL. Stir at 25℃ for 30min, centrifuge at 8000rpm for 10min, discard the precipitate, adjust the pH of the supernatant to 7.0, centrifuge again to discard the precipitate, retain the supernatant, adjust the pH to 5.5, and concentrate under reduced pressure until the solid content is 50%, obtaining a deproteinized fruiting body concentrate. Mix the deproteinized fruiting body concentrate with microcrystalline cellulose at a mass ratio of 1:0.3, and spray dry with air inlet at 60℃ and air outlet at 42℃ to obtain deproteinized Ganoderma lucidum powder with a protein residue of 0.6% and a polysaccharide retention rate of 97%.

[0053] S2, weigh 400g of deproteinized Ganoderma lucidum powder, 35g of croscarmellose sodium, 55g of croscarmellose, 40g of microcrystalline cellulose, 467g of mannitol and 3g of polysorbate 80, put them into a V-type mixer and mix at 20 rpm for 15 minutes to obtain a mixed powder.

[0054] S3: 10.0g of tannic acid and 20.9g of hydroxypropyl-β-cyclodextrin inclusion complex were dissolved in 300mL of 63% ethanol to prepare an inclusion complex solution; 1000g of the mixed powder from S2 was placed in a fluidized bed, and the above inclusion complex solution was uniformly sprayed in at an atomization pressure of 0.3MPa, while controlling the material temperature at 25℃ throughout. After spraying, the moisture content of the material was measured to be 12%, resulting in a soft material.

[0055] S4. The soft material is fed into the hopper of a granulator (1.0 mm / 18 mesh), extruded and sieved at 100 rpm, and then placed in a fluidized bed dryer. It is initially dried at 55°C to a moisture content of 8.0%, yielding approximately 1010 g of initial granules. 30.3 g of ferric ammonium citrate is weighed and dissolved in 70 mL of 4°C pure water. The solution is stirred until completely dissolved to prepare a 30% (w / w) ferric ammonium citrate solution, which is maintained at 4°C. The initial granules are placed in a fluidized bed, and the prepared ferric ammonium citrate solution is sprayed in at an atomizing pressure of 0.15 MPa. The solution is then flash-evaporated at 70°C for 3 min, and finally dried at 60°C to a moisture content of 2.5%, yielding final granules. 10.1 g of magnesium stearate (1.0% of the final granule mass) is weighed as a lubricant and mixed with the final granules in a mixing drum. After standing in a sealed container for 30 min, the mixture is compressed into tablets to obtain Ganoderma lucidum flash-dissolving tablets.

[0056] Example 4 A method for preparing Ganoderma lucidum flash-dissolving tablets includes the following steps: S1, Raw material pretreatment: Add acetate buffer (pH=4.5) to the ultrafine powder of Ganoderma lucidum fruiting bodies at a material-to-liquid ratio of 1g:15mL. Stir at 25℃ for 30min, centrifuge at 8000rpm for 10min, discard the precipitate, adjust the pH of the supernatant to 7.0, centrifuge again to discard the precipitate, retain the supernatant, adjust the pH to 6.0, and concentrate under reduced pressure until the solid content is 65%, obtaining a concentrated solution. Mix the concentrated solution with microcrystalline cellulose at a mass ratio of 1:0.3, and spray dry with air inlet at 60℃ and air outlet at 42℃ to obtain deproteinized Ganoderma lucidum powder with a protein residue of 0.5% and a polysaccharide retention rate of 98%.

[0057] S2, weigh 400g of deproteinized Ganoderma lucidum powder, 35g of croscarmellose sodium, 55g of croscarmellose, 40g of microcrystalline cellulose, 467g of mannitol and 3g of polysorbate 80, put them into a V-type mixer and mix at 20 rpm for 15 minutes to obtain a mixed powder.

[0058] S3: 12.0g of tannic acid and 29.8g of hydroxypropyl-β-cyclodextrin inclusion complex were dissolved in 300mL of 65% ethanol to prepare an inclusion complex solution; 1000g of the mixed powder from S2 was placed in a fluidized bed, and the above inclusion complex solution was uniformly sprayed in at an atomization pressure of 0.3MPa, while controlling the material temperature at 30℃ throughout. After spraying, the moisture content of the material was measured to be 15%, resulting in a soft material.

[0059] S4. The soft material is fed into the hopper of a granulator (1.0 mm / 18 mesh), extruded and sieved at 100 rpm, and then placed in a fluidized bed dryer. It is initially dried at 55°C to a moisture content of 8.0%, yielding approximately 1010 g of initial granules. 30.3 g of ferric ammonium citrate is weighed and dissolved in 70 mL of 4°C pure water. The solution is stirred until completely dissolved to prepare a 30% (w / w) ferric ammonium citrate solution, which is maintained at 4°C. The initial granules are placed in a fluidized bed, and the prepared ferric ammonium citrate solution is sprayed in at an atomizing pressure of 0.15 MPa. The solution is then flash-evaporated at 70°C for 3 min, and finally dried at 60°C to a moisture content of 2.5%, yielding final granules. 10.1 g of magnesium stearate (1.0% of the final granule mass) is weighed as a lubricant and mixed with the final granules in a mixing drum. After standing in a sealed container for 30 min, the mixture is compressed into tablets to obtain Ganoderma lucidum flash-dissolving tablets.

[0060] The molar ratio of tannic acid to hydroxypropyl-β-cyclodextrin in Examples 1 to 4 above was calculated based on a molecular weight of 1701 g / mol for tannic acid and 1400 g / mol for hydroxypropyl-β-cyclodextrin.

[0061] The performance of the deproteinized Ganoderma lucidum powder and Ganoderma lucidum flash-dissolving tablets prepared in Examples 1 to 4 above was tested, and the results are as follows.

[0062] Experiment 1: High-performance liquid chromatography-mass spectrometry analysis of active ingredients in water extracts of Ganoderma lucidum fruiting bodies To detect whether the active ingredients of Ganoderma lucidum fruiting bodies were lost after deproteinization treatment, high performance liquid chromatography-mass spectrometry analysis was performed on its water extract.

[0063] 1. Sample preparation: Weigh an appropriate amount of the deproteinized Ganoderma lucidum powder obtained in Example 1, add deionized water at a material-to-liquid ratio of 1g:30mL, extract in a 90℃ water bath for 2 hours, extract twice, combine the two aqueous extracts, concentrate under reduced pressure at 65℃ to 30% of the original volume, and then dry at 65℃ for 48 hours to obtain the fruiting body aqueous extract.

[0064] The concentration of the fruiting body water extract was diluted to 10 mg / mL with ultrapure water, filtered through a 0.22 μm filter, and then transferred to a liquid chromatography bottle for use in the liquid chromatography system. The ultrapure water filtered through the 0.22 μm filter was used as a blank solution.

[0065] 2. Chromatographic conditions: ACQUITY UPLCTMI-Class and Xevo G3XS liquid chromatography-mass spectrometry (LC-MS) were used for separation. A Waters BEH T3 1.8 m (2.1 mm × 150 mm) column was employed. The mobile phase consisted of 0.1% formic acid (A) and acetonitrile (B) aqueous solution. The flow rate was 0.3 mL / min, the injection volume was 3 µL, and the column temperature was 40 °C. Elution conditions are shown in Table 1.

[0066] Table 1 Gradient elution conditions for high performance liquid chromatography Positive ion mode was used for testing. The sampled mass range was 50~1200 Da. The parameter settings were as follows: capillary voltage 3kV; ion source temperature 100℃; nebulizer temperature 280℃; nebulizer flow rate 800L / h; cone voltage 40V; collision energy was low / high (6V / 20~30V).

[0067] Using the established chromatographic and mass spectrometric conditions, and comparing with the Waters Traditional Medicine database, the chemical constituents of Ganoderma lucidum fruiting body extract were analyzed using UNIFI 1.9.2 software. Considering both retention time and mass spectrometric data, 18 triterpenoids, 7 esters, 3 nucleosides, 9 sterols, and 12 flavonoids were identified in the aqueous extract of Ganoderma lucidum fruiting body. The triterpenoids included ganoderic acid A, ganoderic acid B, ganoderic acid G, ganoderic acid H, ganoderic acid beta, ganoderic acid delta, ganoderic acid epsilon, ganoderic acid theta, ganoderic acid Z, ganodelactone, and ganode-8-en-ric acid LM2. LM2 and other compounds were detected; sterols included β-sitosterol and 22,23-dihydrostigmasterol; flavonoids included apigenin, pinocembrin, acacetin and their glycoside derivatives; esters included acetylated and malonylated glycosides (such as 6''-O-Malonyldaidzin). In addition, organic acids, alkaloids and polyphenols were also detected.

[0068] Experiment 2. Detection of active ingredient content in Ganoderma lucidum fruiting bodies Detection of polysaccharide content in Ganoderma lucidum fruiting bodies a. Preparation of polysaccharide standard solution Accurately measure anhydrous glucose standard and dissolve it in purified water to prepare a stock solution with a concentration of 0.12 mg / mL. Weigh 0.1 g of analytical grade anthrone and mix it with 100 mL of 80% sulfuric acid to prepare an anthrone sulfate solution as an anthrone colorimetric reagent, and store it protected from light.

[0069] A gradient concentration of 0.2 mL was designed. 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, 1.0 mL, and 1.2 mL of glucose standard solution were added to test tubes, respectively. Distilled water was added to a final volume of 2 mL, and the volume was quickly brought to 8 mL with the prepared anthrone colorimetric reagent. The mixture was then thoroughly mixed to obtain polysaccharide standard sample test solutions with concentrations of 3 μg / mL, 6 μg / mL, 9 μg / mL, 12 μg / mL, 15 μg / mL, and 18 μg / mL. After mixing, the solutions were allowed to stand in the dark for 15 min to allow the colorimetric reaction to occur. The colorimetric solution was then transferred to an ice bath and cooled for 15 min to terminate the reaction. The absorbance was then measured at 625 nm using ultraviolet spectrophotometry.

[0070] Table 2 Detection of glucose standard solution R 2 Correlation coefficient formula: Where R 2 Represents the correlation coefficient of the regression equation. A standard curve was plotted with the anhydrous glucose concentration (X, μg / mL) on the x-axis and absorbance (Y) on the y-axis, as shown below. Figure 1 As shown, the regression equation is Y = 0.044X - 0.024, R0 2 =0.9975 (n=6).

[0071] b. Determination of polysaccharide content in Ganoderma lucidum fruiting bodies Accurately weigh 2.00 g of the Ganoderma lucidum flash-dissolving tablets obtained in Example 1, add 60 mL of deionized water and let it swell at room temperature for 1 h. Connect a reflux condenser and reflux in a water bath at 70±5℃ for 4 h. Filter while hot, add an equal amount of deionized water to the filter residue, and reflux the filter residue again for 3 h. Combine the filtrates obtained from the two processes.

[0072] The collected filtrate was concentrated to a paste-like consistency (approximately 5g) in a 65℃ water bath. 5mL of purified water was added to reconstitute the paste, followed by slow addition of 75mL of ethanol, and the mixture was thoroughly stirred. The solution was then incubated at 4℃ for 12 hours. Centrifugation was performed at 4000 rpm, 25℃, and 10 minutes. The supernatant was discarded, and the precipitate was dissolved in 5mL of 65℃ hot water and brought to a final volume of 50mL with purified water. Centrifugation was repeated under the same conditions. 3mL of the supernatant was taken and brought to a final volume of 25mL with purified water to obtain the Ganoderma lucidum polysaccharide test solution. This solution was diluted a certain factor and allowed to stand in the dark for 15 minutes to allow for color development. The colorimetric solution was then transferred to an ice bath and cooled for 15 minutes to terminate the reaction. Finally, using a UV spectrophotometer, 200μL of the test solution was measured at a wavelength of 625nm, and the absorbance was recorded.

[0073] Based on the obtained standard curve, the sample was diluted 70 times, resulting in an absorbance of 0.698. Substituting this into the standard curve Y = 0.044X - 0.024, the concentration of the diluted sample was found to be 16.4 μg / mL. Therefore, the sample of Ganoderma lucidum flash-dissolving tablets obtained in Example 1 contains approximately 1.43% anhydrous glucose, meeting the requirement in the Chinese Pharmacopoeia that Ganoderma lucidum contains no less than 0.9% Ganoderma lucidum polysaccharides.

[0074] Triterpenoid and sterol content detection a. Preparation of standard solutions Accurately weigh ursolic acid standard and dissolve it in methanol to prepare a 0.2 mg / mL standard solution. Accurately weigh 500 mg of vanillin, dissolve it in glacial acetic acid, and dilute to a 10 mL amber volumetric flask. Mix well to obtain a 50 mg / mL colorimetric working solution, which should be prepared immediately before use.

[0075] The indicator was prepared by mixing the colorimetric working solution and perchloric acid in a volume ratio of 1:4.

[0076] Five groups of ursolic acid standard solutions were prepared by transferring 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, and 0.5 mL into test tubes and evaporating to dryness. A total of 1 mL of indicator was added. The reaction system was then transferred to a water bath and heated at 70 °C for 15 min, followed by an ice bath to terminate the reaction for 5 min. Ethyl acetate was added to each of the different experimental levels of the reaction system to bring the volume to 5 mL. After mixing, the absorbance was measured at 550 nm.

[0077] Table 3 Detection of ursolic acid standard solution A standard curve was plotted with ursolic acid concentration (X, μg / mL) on the x-axis and absorbance (Y) on the y-axis, as shown below. Figure 2 As shown, the regression equation is Y = 0.0088X - 0.0068, R02 =0.9989 (n=5).

[0078] b. Determination of triterpenoid and sterol content in Ganoderma lucidum samples Take 2g of the powder of Ganoderma lucidum flash-dissolving tablets obtained in Example 1, add 50mL of ethanol, and extract by ultrasonication for 45min (140W, 42kHz, 45℃). After filtration, add ethanol to the filtrate to make up to 100mL and then dilute to a certain multiple (take 1mL and add ethanol to make up to 10mL) to prepare the Ganoderma lucidum sample triterpenoid and sterol test solution. Take 200μL and measure the absorbance at a wavelength of 550nm.

[0079] The obtained absorbance was 0.125; substituting it into the standard curve "Y=0.0088X-0.0068", the concentration of the test solution was found to be 14.98 μg / mL. Therefore, the sample contained approximately 0.75% triterpenes and sterols, meeting the requirement in the 2020 edition of the Chinese Pharmacopoeia that Ganoderma lucidum contains no less than 0.5% triterpenes and sterols.

[0080] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A method for preparing Ganoderma lucidum flash-dissolving tablets, characterized in that, Includes the following steps: Using ultrafine powder of Ganoderma lucidum fruiting bodies as raw material, the protein is removed to obtain a deproteinized fruiting body concentrate. The deproteinized fruiting body concentrate is mixed with microcrystalline cellulose and then spray-dried to obtain deproteinized Ganoderma lucidum powder. The deproteinized Ganoderma lucidum powder is mixed with excipients to obtain a mixed powder. The mixed powder is then atomized and sprayed with an ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin to obtain a soft material. Using the soft material as a matrix, Ganoderma lucidum flash-dissolved tablets were prepared.

2. The preparation method according to claim 1, characterized in that, The specific method for preparing the deproteinized fruiting body concentrate is as follows: add an acetate buffer solution with a pH of 3.5 to 4.5 to the ultrafine powder of Ganoderma lucidum fruiting bodies, stir and centrifuge at room temperature, take the supernatant, adjust the pH to neutral, centrifuge again, retain the supernatant and adjust the pH to 5 to 6, concentrate under reduced pressure to obtain a deproteinized fruiting body concentrate with a solid content of 40% to 65%.

3. The preparation method according to claim 2, characterized in that, The mass ratio of the deproteinized fruiting body concentrate to microcrystalline cellulose is 1:0.2~0.

4.

4. The preparation method according to claim 1, characterized in that, The inlet air temperature of the spray dryer is 55℃~65℃, and the outlet air temperature is 40℃~45℃.

5. The preparation method according to claim 1, characterized in that, Based on the mass of the mixed powder, the amount of tannic acid added is 0.8%~1.2%, and the molar ratio of hydroxypropyl-β-cyclodextrin to tannic acid is 1.5~3:

1.

6. The preparation method according to claim 1, characterized in that, The ethanol concentration in the ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is 60%~65%, and the mass-volume ratio of the mixed powder to the ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is 1g:0.25mL~0.35mL.

7. The preparation method according to claim 1, characterized in that, When the ethanol inclusion complex solution of tannic acid and hydroxypropyl-β-cyclodextrin is atomized and sprayed into the mixed powder, the temperature of the mixed powder is controlled at 18℃~30℃ and the moisture content is 8%~15%.

8. The preparation method according to claim 1, characterized in that, The soft material is granulated, sized, and tableted to obtain Ganoderma lucidum flash-dissolving tablets. During granulation, an aqueous solution of ferric ammonium citrate with a mass concentration of 25% to 30% at 1℃ to 4℃ is sprayed onto the initial granules obtained after granulation. The mixture is then flash-evaporated at 65℃ to 75℃ for 2 to 4 minutes and finally dried at 50℃ to 60℃ until the moisture content is 2.0% to 3.0%, thus obtaining final granules. During tableting, magnesium stearate with a mass ratio of 0.5% to 1% is added as a lubricant based on the mass of the final granules and mixed with the final granules. The mixture is then allowed to stand and tableted.

9. The preparation method according to claim 1, characterized in that, The excipients include a combined disintegrant consisting of croscarmellose sodium, croscarmellose and microcrystalline cellulose, mannitol filler and surfactant polysorbate 80.

10. A Ganoderma lucidum flash-dissolving tablet prepared by the preparation method according to any one of claims 1 to 9.

Images