Extraction method of ginkgo biloba biflavones, product and application thereof

Abstract

The application discloses a ginkgo biloba biflavonoid extraction method and products and applications thereof, belongs to the technical field of small molecule extraction, and is used for solving the technical problem that the purity and processability of ginkgo biloba biflavonoid need to be further improved in the prior art. The application takes the selective adsorption of phenylboronic acid solidified silica gel, the molecular complexing of L-arginine, and the construction of a double-layer solubilization auxiliary material as core technical paths, realizes efficient conversion of the whole process from ginkgo biloba biflavonoid extraction and purification to tablet forming, removes impurities through specific adsorption, improves dispersibility through molecular complexing, and jointly controls drug release and structure stability through a rapid disintegration solubilization system and a stable solubilization system. The obtained tablet has high purity, fast disintegration, sufficient dissolution, and excellent long-term storage performance, and comprehensively improves the processability and bioavailability of ginkgo biloba biflavonoid in solid preparations.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of small molecule extraction, in particular to a ginkgo biloba biflavonoid extraction method, product and application thereof. BACKGROUND

[0002] Ginkgo biloba biflavonoids are derived from flavonoid glycosides active ingredients in ginkgo leaves, which are often obtained in dispersion by water-alcohol system extraction or ionic liquid, deep eutectic solvent assisted extraction, and then combined with membrane separation, resin adsorption and other methods to improve purity. The methods are mostly aimed at improving the recovery rate and impurity removal efficiency of flavonoids. The obtained products are mainly used in health products, functional foods and traditional Chinese medicine solid preparations. The market demand for high-purity and high-stability raw materials is gradually increasing.

[0003] In terms of preparation application, ginkgo biloba biflavonoids are prone to aggregation between molecules and have strong hydrophobicity. In the processes of tabletting, granulation, spray drying and the like, poor flowability, caking and difficult disintegration often occur. In order to improve the processability, sugar alcohols, cyclodextrins or polysaccharide excipients are often introduced to improve the dispersion and dissolution performance, so that they can adapt to various dosage forms such as tablets, granules and instant powders, and gradually form the application and development direction focusing on solubilization, molding and stabilization treatment.

[0004] At present, in the extraction and purification process of ginkgo biloba biflavonoids, non-selective adsorption or solvent extraction methods are mostly used, which are difficult to effectively distinguish the target flavonoids from similar impurities such as polyphenols and sugars, resulting in insufficient product purity and component stability. Some methods need to rely on strong acid, strong base or organic solvent to adjust the system, which is easy to cause partial degradation or conversion of flavonoid structure, affecting its antioxidant activity and subsequent processing performance. At the same time, the residual impurities and solvents in the extraction product may also have adverse effects on taste, color and safety;

[0005] In the preparation process, ginkgo biloba biflavonoids are prone to form hydrophobic aggregates between molecules, and the powder flowability is poor. After drying, it is easy to caking. The traditional excipient system is difficult to balance the disintegration rate and structural stability. The tablets often have problems such as hardening, delayed dissolution or content reduction during storage. In addition, the lack of carrier system with solubilization and molding functions limits the dissolution efficiency of insoluble flavonoids in solid preparations and the application potential in high-efficiency oral preparations. SUMMARY

[0006] The purpose of the present application is to provide a ginkgo biloba biflavonoid extraction method, product and application thereof, which solves the technical problem that the purity and processability of ginkgo biloba biflavonoids need to be further improved in the prior art.

[0007] The purpose of the present application can be achieved by the following technical solution: a ginkgo biloba biflavonoid extraction method, comprising the following steps:

[0008] S1, taking D50 =100μm of ginkgo biloba leaves powder and deep eutectic solvent were placed in an ultrasonic instrument, and after adding deionized water and pectinase and cellulase, the ultrasonic instrument was heated to 40-50℃, and ultrasonic extraction was carried out for 30 min, and then ginkgo biloba biflavonoid dispersion was obtained by post-treatment;

[0009] S2, the ginkgo biloba biflavonoid dispersion was added to the reaction kettle, and after adjusting the pH of the reaction system to 8.5 with saturated sodium bicarbonate aqueous solution, phenylboronic acid solidified silica gel adsorbent was added, stirred at room temperature for 30 min, then carbon dioxide gas was introduced, and ginkgo biloba biflavonoids were obtained by post-treatment.

[0010] The reaction principle for preparing L-arginine complex ginkgo biloba biflavonoids is as follows:

[0011] First, under the action of deep eutectic solvent and complex enzyme system, the cell wall structure of ginkgo biloba leaves is destroyed, and pectinase and cellulase cooperate to crack the plant polysaccharide matrix, and the strong hydrogen bond network of deep eutectic solvent is used to make the bound flavonoids in ginkgo biloba leaves fully released and form a stable dispersion system. This stage is essentially achieved by solventization and biological catalysis to realize efficient dissociation and transfer of flavonoid glycosides;

[0012] Subsequently, by adjusting the pH of the system and introducing phenylboronic acid solidified silica gel adsorbent, the reversible borate ester bond between the phenylboronic acid group and the ortho-diphenol structure in the flavonoid molecule is used to realize the specific selective adsorption of flavonoid compounds, and at the same time, by introducing carbon dioxide to adjust the pH to neutral, the borate ester bond is destroyed to realize mild desorption, thereby obtaining a high-purity ginkgo biloba biflavonoid purification solution;

[0013] Overall, through multi-stage regulation of physical and chemical interactions, this process realizes the whole process of green transformation from plant tissue wall breaking, targeted enrichment to molecular complex stabilization, providing a feasible chemical basis for the efficient extraction and functional utilization of flavonoids.

[0014] Further, in step S1, the amount ratio of ginkgo biloba leaf powder, deep eutectic solvent, deionized water, pectinase and cellulase is 10g:15-20g:2-3mL:0.05g:0.03g, wherein the deep eutectic solvent is obtained by mixing choline chloride and lactic acid in an amount ratio of 1g:2g, and the post-treatment includes: after ultrasonic completion, the temperature of the ultrasonic instrument is reduced to room temperature, the reaction liquid is filtered to collect the filtrate, and the ginkgo biloba biflavonoid dispersion is obtained;

[0015] Further, in step S2, the amount ratio of the ginkgo biloba biflavonoid dispersion and the phenylboronic acid solidified silica gel adsorbent is 25-30g:1g, and the post-treatment includes: after standing for 10 min, the reaction liquid is filtered to collect the filtrate, and then the filtrate is transferred to a rotary evaporator with a temperature of 80℃, and the pressure is reduced to distill until no liquid is collected, to obtain ginkgo biloba biflavonoids;

[0016] Further, in step S2, the preparation method of the phenylboronic acid solidified silica gel adsorbent is as follows: anhydrous ethanol, 3-aminophenylboronic acid, triethylamine and isocyanic acid propyl triethoxy silane are added into a reaction kettle, after stirring at room temperature for 30-40 min, silica gel, deionized water and glacial acetic acid are continuously added into the reaction kettle, the temperature of the reaction kettle is increased to reflux, and the reaction kettle is kept at reflux for 3-4 h, and the phenylboronic acid solidified silica gel adsorbent is obtained after post-treatment.

[0017] The reaction principle for preparing the phenylboronic acid solidified silica gel adsorbent is as follows:

[0018] Firstly, the amino group in the 3-aminophenylboronic acid molecule reacts with the isocyanic acid group (-NCO) in the isocyanic acid propyl triethoxy silane molecule to generate a stable urea bond connector, so that the phenylboronic acid group is coupled with the organic silane coupling agent containing a hydrolysable silane end group to form an organic intermediate; then, the intermediate is hydrolyzed in an ethanol-water system under acidic conditions, the triethoxy silane group is hydrolyzed to generate silanol (-Si-OH), and further dehydrated and condensed with the hydroxyl group (≡Si-OH) existing on the surface of the silica gel to generate a firm Si-O-Si covalent bond, so that the phenylboronic acid functional group is covalently fixed on the surface of the silica gel;

[0019] In the whole process, the organic end of the silane (the phenylboronic acid group connected by the urea bond) endows the material with specific chemical recognition function, and the inorganic end (Si-O-Si) ensures the stable combination with the silica gel matrix, and finally the obtained adsorbent contains the complete reserved -B(OH)2 group, which can form specific complex with compounds having ortho-diphenol or similar structure (such as flavones and polyphenols) through reversible borate ester bond, so as to realize the selective adsorption and mild desorption of target molecules.

[0020] Further, the amount ratio of anhydrous ethanol, 3-aminophenylboronic acid, triethylamine, isocyanic acid propyl triethoxy silane, silica gel, deionized water and glacial acetic acid is 50-60 mL: 1 g: 0.2 mL: 3-4 g: 10 g: 0.1 mL: 0.02 mL, the pore size of the silica gel is 60-80 A, the particle size is 40-60 μm, and the post-treatment includes: after the reflux is completed, the temperature of the reaction liquid is reduced to room temperature, the reaction liquid is filtered to collect the filter cake, the filter cake is washed with anhydrous ethanol and deionized water for 3-5 times, then the filter cake is transferred to a drying box with a temperature of 80°C for vacuum drying until room temperature, and the phenylboronic acid solidified silica gel adsorbent is obtained.

[0021] The application further discloses a preparation method of the ginkgo biloba biflavonoid product, which comprises the following steps: adding L-arginine, ginkgo biloba biflavonoid and 50wt% ethanol aqueous solution into a reaction kettle, stirring at room temperature for 15-20 minutes, and obtaining the L-arginine-compounded ginkgo biloba biflavonoid through post-treatment, wherein the dosage ratio of the ginkgo biloba biflavonoid purified solution and the L-arginine is 3-4g:0.1g:30mL.

[0022] The principle of preparing the L-arginine-compounded ginkgo biloba biflavonoid is as follows:

[0023] In a neutral to slightly alkaline environment, the L-arginine is introduced, and the cationic characteristics of the guanidino group and the amino group in the molecule are utilized to form an electrostatic interaction, a hydrogen bond combination and a π-π stacking effect between the phenolic hydroxyl group or the carbonyl group of the flavonoid molecule, so that a stable non-covalent complex system is formed. The complex structure not only improves the water solubility and the thermal stability of the flavonoid molecule, but also improves the dispersibility and the bioavailability of the flavonoid molecule in subsequent drying and preparation.

[0024] Further, the dosage ratio of the ginkgo biloba biflavonoid purified solution and the L-arginine is 20-25g:0.1g, and the post-treatment comprises the following steps: after the stirring is completed, the reaction liquid is concentrated to 6-8wt%, and then the L-arginine-compounded ginkgo biloba biflavonoid is obtained through spray drying under the conditions that the inlet air temperature is 145 DEG C, the outlet air temperature is 80 DEG C, the feeding rate is 3-5mL / min, and the atomization air pressure is 0.5-0.7MPa.

[0025] The application further discloses an application of the ginkgo biloba biflavonoid product, and the ginkgo biloba biflavonoid product is applied to the preparation of ginkgo biloba biflavonoid tablets.

[0026] Further, the ginkgo biloba biflavonoid tablet comprises the following raw materials in weight parts: 20 parts of L-arginine-compounded ginkgo biloba biflavonoid, 25-30 parts of fast-disintegrating and solubilizing auxiliary materials, 40-50 parts of stable and solubilizing auxiliary materials, 2-4 parts of crosslinked sodium carboxymethyl cellulose and 1 part of magnesium stearate.

[0027] Further, the preparation method of the fast-disintegrating and solubilizing auxiliary materials comprises the following steps:

[0028] A1, mannitol, hydroxypropyl cellulose, arginine bicarbonate and deionized water are added into a reaction kettle and stirred, and then the porous framework particles are obtained through spray drying under the conditions that the inlet air temperature is 150 DEG C, the outlet air temperature is 75 DEG C, the feeding rate is 3-5mL / min, and the atomization air pressure is 0.5-0.7MPa;

[0029] A2, tartaric acid, ethyl cellulose and anhydrous ethanol are added into a reaction kettle and stirred uniformly to prepare a spraying liquid, the spraying liquid is uniformly sprayed on the surface of the porous framework particles by using a fluidized bed, the film is thickened by 8-10%, and then the fast-disintegrating and solubilizing auxiliary materials are obtained through post-treatment.

[0030] The reaction principle for preparing the rapid disintegration and solubility enhancer is as follows:

[0031] In the system, mannitol, as a hydrophilic skeleton material, forms a hydrogen bond association structure with hydroxypropyl cellulose. Through the rapid solvent evaporation and phase separation effect in the spray drying process, a porous particle structure with regular channels and large specific surface area is generated, providing a physical basis for subsequent rapid disintegration and solubility; arginine bicarbonate decomposes and releases trace amounts of CO2 and water molecules during the drying process, while introducing cationic groups, which not only form micropores in the particle interior and improve porosity, but also impart a certain alkalinity and hydrophilicity to the particle surface, helping to improve the subsequent drug wetting and dissolution rate; hydroxypropyl cellulose, as a binding and film-forming aid, makes mannitol and L-arginine salt complex form a stable dispersion system, preventing particle agglomeration and enhancing structural strength;

[0032] In the subsequent coating stage, ethyl cellulose, as a hydrophobic film-forming material, forms a weak acid-hydrophobic polymer composite film with tartaric acid: tartaric acid interacts with ethyl cellulose molecular chains through hydrogen bonds to adjust the hydrophilicity and disintegrability of the film layer, so that the coating can rapidly swell and crack upon contact with water, promoting water penetration into the skeleton interior. This composite film can not only maintain the stability of the particle structure during normal temperature storage, but also rapidly disintegrate upon contact with water, achieving rapid drug release and high efficiency dissolution;

[0033] Overall, the formation process of the enhancer is a structure self-organization process driven by hydrogen bond assembly, gas poration, phase separation pore formation, and surface functionalization coating. Through this multi-dimensional synergistic mechanism, the obtained rapid disintegration and solubility enhancer simultaneously possesses high porosity, good disintegrability, and excellent water-wetting and solubilizing properties, providing a physicochemical basis for the rapid release of poorly soluble active ingredients in solid preparations.

[0034] Further, in step A1, the amount ratio of the mannitol, hydroxypropyl cellulose, arginine bicarbonate, and deionized water is 7-8 g: 1 g: 1 g: 20 mL;

[0035] Further, in step A2, the amount ratio of the tartaric acid, ethyl cellulose, and anhydrous ethanol is 8-10 g: 3 g: 200 mL, and the post-treatment includes: transferring the material to a drying box with a temperature of 40℃ and vacuum drying to a constant weight to obtain the rapid disintegration and solubility enhancer.

[0036] Further, the preparation method of the stable solubility enhancer comprises the following steps:

[0037] B1, corn starch and deionized water are added to the reaction kettle and stirred, and the temperature of the reaction kettle is raised to 70-80℃, and after 5-10 min of heat preservation and stirring, the pre-gelatinized starch is obtained by post-treatment;

[0038] B2, β-cyclodextrin, microcrystalline cellulose and pregelatinized starch were added into the reaction kettle and stirred uniformly, then anhydrous ethanol and deionized water were added for wet granulation, after granulation, D 50 =300pm particles, and after post-processing, a solubilizing excipient precursor was obtained;

[0039] B3, at 45°C, 0.5wt% citric acid aqueous solution was uniformly sprayed on the surface of the solubilizing excipient precursor by fluidized bed, the film weight was increased by 4-5%, and after post-processing, a stable solubilizing excipient was obtained.

[0040] The reaction principle for preparing the stable solubilizing excipient is:

[0041] Firstly, after heat treatment, corn starch is partially gelatinized and the molecular rearrangement occurs, the crystal structure is destroyed, and the branched starch molecular chain is unfolded in the water phase to form an amorphous three-dimensional gel network, which gives the system good cohesiveness and forming property, and also serves as the subsequent structural framework and bonding matrix;

[0042] Subsequently, in the wet granulation system, β-cyclodextrin, microcrystalline cellulose and pregelatinized starch form a stable multiphase composite network through intermolecular hydrogen bonding and van der Waals forces: β-cyclodextrin provides a hydrophobic inclusion cavity, which can include hydrophobic or poorly soluble components through host-guest interaction, thereby significantly improving the solubilizing capacity of the system; microcrystalline cellulose as an inert framework material provides mechanical support and controls the pore structure of the granules, making the system have certain fluidity and compression stability; pregelatinized starch is cross-linked to form a continuous phase structure after drying, which makes cyclodextrin and cellulose uniformly dispersed in the granule matrix, preventing component migration and phase separation, thereby constructing a porous precursor particle with strength and solubilizing property;

[0043] In the final surface modification stage, a layer of weakly acidic film containing carboxyl groups is formed on the surface of the granules by spraying citric acid solution, and citric acid molecules can partially esterify or hydrogen bond with the hydroxyl groups in the granules, thereby forming a dense and polar-regulated protective layer on the surface; this film layer can not only stabilize the internal structure of the granules through weak acid-hydroxyl interaction to prevent moisture absorption and collapse, but also can quickly swell and disintegrate during use, promoting water infiltration and activating the inclusion of cyclodextrin, achieving a dynamic balance between structural stability and rapid solubilizing property;

[0044] Overall, the formation of the stable solubilizing excipient relies on a multi-level synergistic mechanism of starch thermal denaturation, cyclodextrin molecular inclusion, cellulose framework support and citric acid surface regulation. This system realizes the unity of structural stability and functional optimization through intermolecular interaction and interface energy regulation, thereby giving the excipient excellent physicochemical stability, solubilizing capacity and molding adaptability, providing an efficient solubilization and carrier basis for the formulation of poorly soluble active substances.

[0045] Further, in step B1, the ratio of corn starch and deionized water is 1g:3-4g, and the post-treatment includes: spreading the reaction system on a stainless steel plate, drying at 60 DEG C hot air until the water content is less than or equal to 5%, and then passing through an 80-mesh sieve to obtain pregelatinized starch;

[0046] Further, in step B2, the ratio of beta-cyclodextrin, microcrystalline cellulose, pregelatinized starch, anhydrous ethanol and deionized water is 5g:12-15g:1g:10mL:30mL, and the post-treatment includes: transferring the particles to a drying oven at a temperature of 40-45 DEG C and vacuum drying until the moisture content is less than or equal to 4% to obtain a solubilizing adjuvant precursor;

[0047] Further, in step B3, the post-treatment includes: transferring the material to a drying oven at a temperature of 80 DEG C and vacuum drying until the constant weight is obtained to obtain a stable solubilizing adjuvant.

[0048] The present application has the following beneficial effects:

[0049] 1、In the extraction process of ginkgo biloba biflavones, the present application uses a deep eutectic solvent to form a dense hydrogen bond network, which can effectively destroy the cell structure of ginkgo leaves, promote the full release of flavonoid compounds, and inhibit the co-dissolution of impurities; the active boron hydroxyl functional groups on the surface of the phenylboronic acid solidified silica gel adsorbent can selectively bind with the ortho-diphenol structure in the flavonoid molecule, realizing specific recognition and adsorption separation through reversible borate ester bond action; the organic connection of the two systems in the extraction and purification stages enables the flavonoid components to be highly selectively enriched, and the impurities are effectively removed; the whole process is mild and controllable, avoiding the thermal degradation and structural damage of active ingredients; through the synergistic regulation of intermolecular interaction, the present application forms a positive synergistic effect in terms of extraction efficiency, purity control and system stability, significantly improving the purification quality and process stability of ginkgo biloba biflavones.

[0050] 2、The guanidine group and amino group in the L-arginine molecule form hydrogen bonds and electrostatic complexes with biflavone molecules, significantly improving the water solubility and dispersion stability of the biflavones, laying a foundation for the uniform distribution and rapid dissolution of the drug in the solid system; the rapid disintegration and solubilizing adjuvant rapidly absorbs water and swells to form liquid channels through the synergistic action of the porous skeleton and hydrophilic groups after encountering liquid, accelerating the disintegration of the tablets; the stable solubilizing adjuvant prevents ingredient migration and structure collapse while maintaining the mechanical strength of the tablets through cyclodextrin inclusion and cellulose skeleton support, forming a multi-level network of synergistic solubilization, rapid disintegration and structural stability at the micro level, so that the tablets can achieve rapid disintegration and release and high-efficiency dissolution while having appropriate hardness, significantly improving the dissolution behavior and use stability of ginkgo biloba biflavones.

[0051] 3、The guanidine group and the amino group in the L-arginine molecule of the application can form hydrogen bonds and electrostatic complexes with the ginkgo biloba biflavone molecule, significantly improving the hydrophilicity and dispersion stability thereof, and inhibiting molecular aggregation and oxidative degradation at high temperatures, thereby enhancing the thermal stability of the system; the rapid disintegration and solubilization adjuvant expands rapidly after absorbing water, forming porous channels to accelerate the disintegration and release of the tablet; the cyclodextrin in the stable and solubilization adjuvant cooperates with the microcrystalline cellulose to construct a support network, so that the tablet maintains mechanical strength while maintaining uniform distribution of components and structural integrity, and the three form a complementary synergistic effect at the molecular and microscopic levels, achieving efficient dissolution and rapid disintegration, and imparting excellent storage temperature stability to the tablet, which can still maintain structural stability, stable disintegration performance and effective ingredient content under a relatively high temperature and humidity environment, ensuring the consistency and long-term controllability of the quality of the preparation. BRIEF DESCRIPTION OF DRAWINGS

[0052] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or the prior art description will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0053] Figure 1 The infrared absorption spectrum image of the phenylboronic acid cured silica gel adsorbent prepared in Example 3;

[0054] Figure 2 The infrared absorption spectrum image of the L-arginine complex ginkgo biloba biflavone prepared in Example 3;

[0055] Figure 3 The infrared absorption spectrum image of the rapid disintegration and solubilization adjuvant prepared in Example 6;

[0056] Figure 4 The infrared absorption spectrum image of the stable and solubilization adjuvant prepared in Example 6. DETAILED DESCRIPTION

[0057] The technical solutions of the present application will be described in detail below in conjunction with the embodiments, obviously, the described embodiments are only some of the embodiments of the present application, not all. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0058] In the present application, the silica gel used is purchased from Biyun Tian Biotechnology, with the item number Y187016; the cross-linked sodium carboxymethyl cellulose used is purchased from Xi'an Tianzheng Pharmaceutical Auxiliary Material Co., Ltd.; the cellulase used is purchased from Shanghai Macklin Biochemical Technology Co., Ltd., with the item number C758963; the pectinase used is purchased from Shanghai Macklin Biochemical Technology Co., Ltd., with the item number M6346; the hydroxypropyl cellulose used is purchased from Shanghai Macklin Biochemical Technology Co., Ltd., with the item number L823212; the ethyl cellulose used is purchased from Shanghai Macklin Biochemical Technology Co., Ltd., with the item number E809014; the corn starch used is purchased from Shanghai Macklin Biochemical Technology Co., Ltd., with the item number 768498; the microcrystalline cellulose used is purchased from Shanghai Macklin Biochemical Technology Co., Ltd., with the item number M909921; the cross-linked sodium carboxymethyl cellulose used is purchased from Shanghai Yuanye Biotechnology Co., Ltd., with the item number S25102; the magnesium stearate used is purchased from Shanghai Yuanye Biotechnology Co., Ltd., with the item number S52902.

[0059] Example 1

[0060] The present embodiment provides a preparation method of ginkgo biloba biflavonoid product, comprising the following steps:

[0061] Step 1, preparation of phenylboronic acid solidified silica gel adsorbent

[0062] Take: 500.0 mL of anhydrous ethanol, 10.0 g of 3-aminophenylboronic acid, 2.0 mL of triethylamine and 30.0 g of propyl triethoxy silane isocyanate into the reaction kettle, stir for 30 min at room temperature, then add 100.0 g of silica gel with a pore size of 60A and a particle size of 40 μm, 1.0 mL of deionized water and 0.2 mL of glacial acetic acid into the reaction kettle, then increase the temperature of the reaction kettle to reflux, and keep the temperature at reflux for 3 h. After the reflux is completed, the temperature of the reaction solution is reduced to room temperature, the reaction solution is filtered to collect the filter cake, the filter cake is washed with anhydrous ethanol and deionized water for 3 times, then the filter cake is transferred to a drying box with a temperature of 80°C and vacuum dried to room temperature to obtain the phenylboronic acid solidified silica gel adsorbent.

[0063] Step 2, preparation of ginkgo biloba biflavonoid dispersion

[0064] Take: 100.0 g of choline chloride and 200.0 g of lactic acid to obtain a deep eutectic solvent;

[0065] Take: 100.0 g of D 50Ginkgo biloba powder with particle size of 100 μm and 150.0 g deep eutectic solvent were placed in an ultrasonic instrument, 20.0 mL of deionized water and 0.5 g of pectinase and 0.3 g of cellulase were added, the ultrasonic instrument was heated to 40℃, and ultrasonic extraction was performed for 30 min. After ultrasonic extraction, the temperature of the ultrasonic instrument was reduced to room temperature, the reaction solution was filtered, and the filtrate was collected to obtain a ginkgo bilobate dispersion.

[0066] Step 3, preparation of ginkgo bilobate purification solution

[0067] Weighing: 250.0 g of ginkgo bilobate dispersion was added to the reaction kettle, and 10.0 g of phenylboronic acid solid silica gel adsorbent was added after the pH of the reaction system was adjusted to 8.5 using saturated sodium bicarbonate aqueous solution. Stirring was performed at room temperature for 30 min, then carbon dioxide gas was introduced to adjust the pH of the reaction system to 6.0, and the reaction was allowed to stand for 10 min. The reaction solution was filtered, and the filtrate was transferred to a rotary evaporator with a temperature of 80℃. The filtrate was distilled under reduced pressure until no liquid was collected, and ginkgo bilobate was obtained.

[0068] Step 4, preparation of L-arginine complex ginkgo bilobate

[0069] Weighing: 1.0 g of L-arginine, 40.0 g of ginkgo bilobate, and 300 mL of 50 wt% ethanol aqueous solution were added to the reaction kettle, and stirring was performed at room temperature for 15 min. After the reaction was completed, the reaction solution was concentrated to 6 wt%, and spray drying was performed under the conditions of inlet air temperature of 145℃, outlet air temperature of 80℃, feed rate of 3 mL / min, and atomization gas pressure of 0.5 MPa to obtain L-arginine complex ginkgo bilobate.

[0070] Example 2

[0071] The present embodiment provides a method for preparing ginkgo bilobate product, which comprises the following steps:

[0072] Step 1, preparation of phenylboronic acid solid silica gel adsorbent

[0073] Weighing: 600.0 mL of anhydrous ethanol, 10.0 g of 3-aminophenylboronic acid, 2.0 mL of triethylamine, and 40.0 g of propyl triethoxysilane isocyanate were added to the reaction kettle, and stirring was performed at room temperature for 40 min. Then, 100.0 g of silica gel with a pore size of 80 A and a particle size of 60 μm, 1.0 mL of deionized water, and 0.2 mL of glacial acetic acid were added to the reaction kettle, and the temperature of the reaction kettle was increased to reflux. The reaction was maintained at reflux for 4 h. After the reaction was completed, the temperature of the reaction solution was reduced to room temperature, the reaction solution was filtered, and the filter cake was washed with anhydrous ethanol and deionized water for 5 times. Then, the filter cake was transferred to a drying oven with a temperature of 80℃ and vacuum dried to room temperature to obtain a phenylboronic acid solid silica gel adsorbent.

[0074] Step 2, preparation of ginkgo bilobate dispersion

[0075] Take: 100.0 g of choline chloride and 200.0 g of lactic acid to prepare a deep eutectic solvent;

[0076] Take: 100.0 g of D 50 =100 μm ginkgo biloba powder and 200.0 g of deep eutectic solvent were placed in an ultrasonic instrument, and after adding 30.0 mL of deionized water and 0.5 g of pectinase and 0.3 g of cellulase, the ultrasonic instrument was heated to 50°C, and ultrasonic extraction was performed for 30 min. After the ultrasonic extraction was completed, the temperature of the ultrasonic instrument was reduced to room temperature, the reaction liquid was filtered to collect the filtrate, and a ginkgo biloba biflavonoid dispersion liquid was obtained.

[0077] Step 3, preparation of ginkgo biloba biflavonoid purification liquid

[0078] Take: 300.0 g of ginkgo biloba biflavonoid dispersion liquid was added to the reaction kettle, and after adjusting the pH of the reaction system to 8.5 using saturated sodium bicarbonate aqueous solution, 10.0 g of phenylboronic acid solid silica gel adsorbent was added, and stirred at room temperature for 30 min. Then, carbon dioxide gas was introduced to adjust the pH of the reaction system to 6.0, and after standing for 10 min, the reaction liquid was filtered to collect the filtrate, and then the filtrate was transferred to a rotary evaporator with a temperature of 80°C. The filtrate was distilled under reduced pressure until no liquid was collected, and ginkgo biloba biflavonoid was obtained.

[0079] Step 4, preparation of L-arginine complex ginkgo biloba biflavonoid

[0080] Take: 1.0 g of L-arginine, 30.0 g of ginkgo biloba biflavonoid, and 300 mL of 50 wt% ethanol aqueous solution were added to the reaction kettle, and stirred at room temperature for 20 min. After the stirring was completed, the reaction liquid was concentrated to 8 wt%, and then spray dried under the conditions of inlet air temperature 145°C, outlet air temperature 80°C, feed rate 5 mL / min, and atomization gas pressure 0.7 MPa to obtain L-arginine complex ginkgo biloba biflavonoid.

[0081] Example 3

[0082] The present embodiment provides a method for preparing ginkgo biloba biflavonoid product, comprising the following steps:

[0083] Step 1, preparation of phenylboronic acid solid silica gel adsorbent

[0084] Take: 550.0 mL of anhydrous ethanol, 10.0 g of 3-aminobenzene boronic acid, 2.0 mL of triethylamine and 36.0 g of isocyanate propyl triethoxysilane into the reaction kettle, after stirring at room temperature for 36 min, continue to add 100.0 g of silica gel with a pore size of 70 A and a particle size of 50 μm, 1.0 mL of deionized water and 0.2 mL of glacial acetic acid into the reaction kettle, then increase the temperature of the reaction kettle to reflux, and keep the temperature at reflux for 4 h. After the reflux is completed, when the temperature of the reaction solution decreases to room temperature, filter the reaction solution to collect the filter cake, wash the filter cake with anhydrous ethanol and deionized water for 4 times, then transfer the filter cake to a drying box with a temperature of 80°C for vacuum drying until room temperature. The benzene boronic acid solidified silica gel adsorbent is obtained.

[0085] The ATR-FTIR spectrum of the obtained benzene boronic acid solidified silica gel adsorbent sample is shown in Figure 1 (4000-400 cm -1 ). The spectrum presents a clear and stable baseline, superimposed with slight high-frequency noise, and the signal-to-noise ratio is good. The reaction sample is a solid silica gel adsorbent with uniform network structure.

[0086] The strong asymmetric stretching peak of Si-O-Si at 1080 cm -1 and the symmetric stretching peak at 800 cm -1 together confirm the silica gel skeleton, and the Si-OH / Si-O-H absorption at 950 cm -1 indicates that the surface residual hydroxyl group still exists; the O-H / N-H wide band and shoulder peak at 3450 / 3330 cm -1 , combined with the obvious urea C=O absorption at 1655 cm -1 and the C-N (urea) at 1250 cm -1 , verify that the urea bond is formed on the surface after the condensation of isocyanate propyl triethoxysilane and 3-aminobenzene boronic acid.

[0087] The aromatic ring part presents a medium intensity C=C band at 1600 / 1510 cm -1 , a weak aromatic C-H absorption at 3030 cm -1 , and an out-of-plane bending signal of the aromatic ring at 770 / 700 cm -1 , indicating that the benzene ring end group is successfully introduced; the boronic acid related vibration appears at 1330 cm -1 (B-O) and 1205 cm -1 (B-O-C / aryl-B environment) at the same time, indicating that the benzene boronic acid functional group is in a covalently fixed / silanized peripheral environment.

[0088] In addition, the aliphatic C-H stretching at 2930 / 2860 cm -1 characterizes the existence of propyl silane segments, and no free isocyanate (~2270 cm -1) and significant organic solvent residue strong peaks, the co-occurrence of the above-mentioned “Si-O-Si main skeleton + urea bond C=O / C-N + benzene ring C=C / out-of-plane bending + B-O / B-O-C” key peak system, and consistent with the expected peak position, verify that the sample is the target adsorbent of phenylboronic acid cured on the surface of silica gel by urea bond-silanization, and the structure construction is consistent with the process expectation.

[0089] Step 2, preparation of ginkgo biflavonoid dispersion

[0090] Take: 100.0 g of choline chloride and 200.0 g of lactic acid to obtain a deep eutectic solvent;

[0091] Take: 100.0 g of D 50 =100 μm ginkgo leaf powder and 180.0 g of deep eutectic solvent into an ultrasonic instrument, and after adding 25.0 mL of deionized water and 0.5 g of pectinase and 0.3 g of cellulase, the ultrasonic instrument is heated to 45℃, and the ultrasonic extraction is kept for 30 min. After the ultrasonic extraction is completed, the temperature of the ultrasonic instrument is reduced to room temperature, the reaction liquid is filtered to collect the filtrate, and the ginkgo biflavonoid dispersion is obtained.

[0092] Step 3, preparation of ginkgo biflavonoid purification liquid

[0093] Take: 270.0 g of ginkgo biflavonoid dispersion into a reaction kettle, and after adjusting the pH of the reaction system to 8.5 with saturated sodium bicarbonate aqueous solution, add 10.0 g of phenylboronic acid solidified silica gel adsorbent, stir at room temperature for 30 min, then pass carbon dioxide gas to adjust the pH of the reaction system to 6.0, stand for 10 min, then filter the reaction liquid to collect the filtrate, and then transfer the filtrate to a rotary evaporator with a temperature of 80℃, and distill under reduced pressure until no liquid is collected. Ginkgo biflavonoids are obtained.

[0094] Step 4, preparation of L-arginine complex ginkgo biflavonoids

[0095] Take: 1.0 g of L-arginine, 36.0 g of ginkgo biflavonoids and 300 mL of 50 wt% ethanol aqueous solution into a reaction kettle, stir at room temperature for 18 min, after the stirring is completed, concentrate the reaction liquid to 7 wt%, and then spray dry under the conditions of inlet air temperature 145℃, outlet air temperature 80℃, feed rate 4 mL / min, and atomization gas pressure 0.6 MPa to obtain L-arginine complex ginkgo biflavonoids.

[0096] The L-arginine complex ginkgo biflavonoids sample obtained by spray drying is tested by ATR-FTIR (4000-400 cm -1 ), as shown in Figure 2 , the spectral baseline is smooth, the noise is slight, and the signal-to-noise ratio is good;

[0097] The spectrum is shown in -1 and 1415 cm-1 present a pair of carboxylate characteristic absorption peaks with a peak interval of about 190 cm -1 , and 1710-1730 cm -1 No free carboxylic acid C=O absorption is observed in the region, indicating that the carboxylic acid groups in the sample have been ionized and formed salt type structure, which is direct evidence of the complexation of L-arginine with ginkgo biloba biflavones;

[0098] 3330 cm -1 Broad band with 3205 cm -1 shoulder peak corresponds to the N-H / O-H absorption of hydrogen bonding, combined with the C=N / C=O complex absorption at 1648 cm -1 and the C-N absorption at 1365 cm -1 , together verify that the guanidino cation has been introduced into the complex system;

[0099] 1648 cm -1 of conjugated carbonyl absorption, 1605 and 1510 cm -1 of aromatic ring C=C vibration, and continuous C-O and C-O-C absorption in the range of 1275-1050 cm -1 , indicating that the biflavone nucleus structure is complete, and its phenolic hydroxyl group participates in ionic or hydrogen bonding;

[0100] 835 and 765 cm -1 of aromatic ring C-H out-of-plane bending vibration signal further confirms the preservation of aromatic structure;

[0101] No -NCO absorption and strong free acid carbonyl peak appears in the spectrum, and there is no significant solvent characteristic band interference. The comprehensive spectral characteristics can be determined that the carboxylate characteristic peak, guanidino absorption band and biflavone skeleton signal exist in the sample at the same time, which fully verifies that the prepared sample is L-arginine complexed with ginkgo biloba biflavones (salt type complex), and its structure meets the characterization requirements of the target product.

[0102] Example 4

[0103] The present embodiment provides a preparation method of ginkgo biloba biflavone tablets, comprising the following steps:

[0104] Step 1, preparation of fast disintegrating and solubilizing adjuvant

[0105] Take 7.0 g of mannitol, 1.0 g of hydroxypropyl cellulose, 1.0 g of arginine bicarbonate and 20.0 mL of deionized water and add them to the reaction kettle for stirring. After uniform stirring, spray drying is carried out under the conditions of inlet air temperature 150℃, outlet air temperature 75℃, feeding rate 3 mL / min, and atomization gas pressure 0.5 MPa to obtain porous skeleton particles;

[0106] Weighing: 8.0 g tartaric acid, 3.0 g ethyl cellulose and 200.0 mL anhydrous ethanol into the reaction kettle, stirring uniformly, then preparing the spraying liquid. The fluidized bed is used to uniformly spray the spraying liquid on the surface of 6.0 g porous framework particles, so that the film weight increases by 8%. After that, the material is transferred to a drying oven with a temperature of 40℃ for vacuum drying until the constant weight, and the rapid disintegration and solubilization adjuvant is obtained.

[0107] Step two, preparation of stable solubilization adjuvant

[0108] Weighing: 10.0 g corn starch and 30.0 g deionized water into the reaction kettle, stirring, and increasing the temperature of the reaction kettle to 70℃. After 5 min of heat preservation and stirring, the reaction system is spread on a stainless steel plate and dried with hot air at 60℃ until the water content is ≤5%. Then, it is sieved through an 80 mesh sieve to obtain pregelatinized starch.

[0109] Weighing: 5.0 g β-cyclodextrin, 12.0 g microcrystalline cellulose and 1.0 g pregelatinized starch into the reaction kettle, stirring uniformly, then adding 10.0 mL anhydrous ethanol and 30.0 mL deionized water for wet granulation. After the granulation is completed, D 50 =300µm particles are obtained. The particles are transferred to a drying oven with a temperature of 40℃ for vacuum drying until the moisture content is ≤4%, and the solubilization adjuvant precursor is obtained.

[0110] At 45℃, the fluidized bed is used to uniformly spray 0.5wt% citric acid aqueous solution spraying liquid on the surface of 10.0 g solubilization adjuvant precursor, so that the film weight increases by 4%. After that, the material is transferred to a drying oven with a temperature of 80℃ for vacuum drying until the constant weight, and the stable solubilization adjuvant is obtained.

[0111] Step three, preparation of ginkgetin tablets

[0112] According to weight parts, 20 parts of L-arginine complex ginkgetin prepared in Example 1, 25 parts of rapid disintegration and solubilization adjuvant, 40 parts of stable solubilization adjuvant, 2 parts of crosslinked sodium carboxymethyl cellulose and 1 part of magnesium stearate are weighed and compressed into shape to obtain ginkgetin tablets.

[0113] Example 5

[0114] This example provides a preparation of ginkgetin tablets, which includes the following steps:

[0115] Step one, preparation of rapid disintegration and solubilization adjuvant

[0116] Weighing: 8.0 g mannitol, 1.0 g hydroxypropyl cellulose, 1.0 g arginine bicarbonate and 20.0 mL deionized water into the reaction kettle, stirring, and then spray drying under the conditions of inlet air temperature 150℃, outlet air temperature 75℃, feeding rate 5 mL / min and atomization gas pressure 0.7 MPa to obtain porous framework particles.

[0117] Weighing: 10.0 g of tartaric acid, 3.0 g of ethyl cellulose and 200.0 mL of anhydrous ethanol are added to the reaction kettle and stirred uniformly to prepare a spraying solution. The spraying solution is uniformly sprayed onto the surface of 6.0 g of porous framework particles using a fluidized bed, and the film is increased by 10% after drying. The material is transferred to a drying oven at a temperature of 40°C and vacuum dried to constant weight to obtain a rapid disintegration and solubilization adjuvant.

[0118] Step two, preparation of stable solubilization adjuvant

[0119] Weighing: 10.0 g of corn starch and 40.0 g of deionized water are added to the reaction kettle and stirred, and the temperature of the reaction kettle is increased to 80°C. After 10 min of incubation and stirring, the reaction system is spread on a stainless steel plate and dried at 60°C with hot air until the water content is ≤5%. After passing through an 80-mesh screen, pre-gelatinized starch is obtained.

[0120] Weighing: 5.0 g of β-cyclodextrin, 15.0 g of microcrystalline cellulose and 1.0 g of pre-gelatinized starch are added to the reaction kettle and stirred uniformly. Then 10.0 mL of anhydrous ethanol and 30.0 mL of deionized water are added for wet granulation. After granulation, D 50 =300µm particles are obtained. The particles are transferred to a drying oven at a temperature of 45°C and vacuum dried to a moisture content of ≤4% to obtain a solubilization adjuvant precursor.

[0121] At 45°C, a 0.5wt% citric acid aqueous solution is uniformly sprayed onto the surface of 10.0 g of the solubilization adjuvant precursor using a fluidized bed, and the film is increased by 5% after drying. The material is transferred to a drying oven at a temperature of 80°C and vacuum dried to constant weight to obtain a stable solubilization adjuvant.

[0122] Step three, preparation of ginkgo biloba biflavonoid tablets

[0123] According to weight parts, 20 parts of L-arginine complex ginkgo biloba biflavonoids prepared in Example 2, 30 parts of rapid disintegration and solubilization adjuvant, 50 parts of stable solubilization adjuvant, 4 parts of cross-linked sodium carboxymethyl cellulose and 1 part of magnesium stearate are pressed into shape to obtain ginkgo biloba biflavonoid tablets.

[0124] Example 6

[0125] This example provides a preparation of ginkgo biloba biflavonoid tablets, comprising the following steps:

[0126] Step one, preparation of rapid disintegration and solubilization adjuvant

[0127] Weighing: 8.0 g of mannitol, 1.0 g of hydroxypropyl cellulose, 1.0 g of arginine bicarbonate and 20.0 mL of deionized water are added to the reaction kettle and stirred, and after being stirred uniformly, spray drying is carried out under the conditions of an air inlet temperature of 150°C, an air outlet temperature of 75°C, a feeding rate of 4 mL / min, and an atomizing gas pressure of 0.6 MPa, to obtain porous framework particles;

[0128] Weighing: 9.0 g of tartaric acid, 3.0 g of ethyl cellulose and 200.0 mL of anhydrous ethanol are added to the reaction kettle and stirred uniformly to prepare a spraying liquid, the spraying liquid is uniformly sprayed onto the surface of 6.0 g of porous framework particles by using a fluidized bed, the film is increased by 9%, and then the material is transferred to a drying box with a temperature of 40°C for vacuum drying to a constant weight, to obtain a rapid disintegration and solubilization auxiliary material.

[0129] The obtained rapid disintegration and solubilization auxiliary material sample is tested by ATR-FTIR (4000-400 cm -1 ), as shown in FIG. 1, the spectrum baseline is smooth, the superposition is slight noise, and the signal resolution is good; Figure 3

[0130] The spectrum shows a wide strong absorption at 3380 cm -1 , which is the superposition of O-H and N-H stretching vibration in mannitol, hydroxypropyl cellulose and arginine bicarbonate, indicating that the system contains multiple hydroxyl and amino groups, and the absorption peaks at 2930 cm -1 and 2880 cm -1 belong to C-H stretching vibration, which is the characteristic signal of ethyl cellulose and organic framework;

[0131] An obvious C=O absorption appears at 1712 cm -1 , which is the esterification or carboxylic acid carbonyl vibration peak of tartaric acid, and is a verification sign of the formation of the coating film, and a medium-intensity band appears near 1600 cm -1 , which belongs to the N-H bending or C=N vibration of L-arginine residue, indicating that the amino component has participated in the complex system;

[0132] The absorption at 1460 cm -1 and 1385 cm -1 corresponds to CH2 bending and C-N / COO - vibration, reflecting the existence of organic salt and amino acid residue, and the continuous C-O-C and C-O stretching absorption in the range of 1265-1025 cm -1 is strong and stable, proving that the polysaccharide ether bond and polyhydroxy framework in the material remain intact;

[0133] The signals at 1080 cm -1 and 1025 cm -1 are particularly significant, which are characteristic peaks of mannitol and hydroxypropyl cellulose, and the signals at 850 cm -1 and 760 cm​-1 The weak peaks are CH bending vibration signals, indicating the presence of a film layer on the surface of ethyl cellulose;

[0134] The absence of obvious free acid carbonyl abnormal peaks and solvent residue peaks in the full spectrum indicates that the system was thoroughly dried and chemically stable. The presence of positional correspondences among the characteristic absorptions confirms that the obtained sample is a rapid-disintegration solubilizing excipient composed of a mannitol-hydroxypropyl cellulose backbone, arginine bicarbonate solubilizing component, and tartaric acid-ethyl cellulose coating membrane. Its infrared spectral characteristics are consistent with the expected structure, proving that the target excipient was successfully prepared.

[0135] Step 2: Preparation of stable solubilizing excipients

[0136] Weigh out 10.0g of corn starch and 36.0g of deionized water and add them to the reaction vessel. Stir the mixture and raise the temperature of the reaction vessel to 75℃. After stirring for 8 minutes, spread the reaction system on a stainless steel plate and dry it with hot air at 60℃ until the moisture content is ≤5%. Then, pass it through an 80-mesh sieve to obtain pregelatinized starch.

[0137] Weigh out 5.0 g β-cyclodextrin, 15.0 g microcrystalline cellulose, and 1.0 g pregelatinized starch, add them to a reaction vessel, stir well, then add 10.0 mL anhydrous ethanol and 30.0 mL deionized water for wet granulation. After granulation, obtain D. 50 Particles with a diameter of 300µm were transferred to a drying oven at 45℃ and vacuum dried until the moisture content was ≤4% to obtain a solubilizing excipient precursor.

[0138] At 45℃, a 0.5wt% citric acid aqueous solution was uniformly sprayed onto the surface of 10.0g of solubilizing excipient precursor using a fluidized bed. After the film weight increased by 5%, the material was transferred to a drying oven at 80℃ and vacuum dried to constant weight to obtain a stable solubilizing excipient.

[0139] The prepared stable solubilizing excipient samples were tested by ATR-FTIR (4000-400 cm⁻¹). -1 ),like Figure 4 As shown, 3380cm -1 The broad and strong OH absorption is a general characteristic of the starch / β-cyclodextrin / microcrystalline cellulose hydrogen bond network, 2930 / 2890 cm⁻¹. -1 For the stretching of the organic framework CH;

[0140] 1716cm -1 The presence of a medium-intensity carbonyl band confirms the presence of the citric acid coating / carbonyl component; 1645 cm⁻¹ -1 For the bending vibration of bound water; 1462, 1424, 1374, 1336 cm -1 CH2 bending and skeletal bands are typical fingerprints of cellulose / starch;

[0141] 1160-995cm -1 Interval continuous strong absorption (including 1160, 1112, 1055, 1030, 995cm -1 ) corresponds to polysaccharide C-O-C / C-O stretching cluster, 897cm -1 The β-glycosidic bond indicating peak is clearly present, proving that the polysaccharide backbone and the cyclodextrin skeleton are maintained; 760cm -1 Weak band is a film / lattice related bending signal, no abnormal solvent strong peak and free salt / amine characteristic peak interference is found in the full spectrum,

[0142] The above key features (co-occurrence of polysaccharide C-O-C / C-O fingerprint cluster + β-glycosidic bond peak, superposition of 1716cm -1 carbonyl band and 3380cm -1 wide O-H) together verify that the sample is composed of a polysaccharide matrix composed of pregelatinized starch-β-cyclodextrin-microcrystalline cellulose and a thin coating film of citric acid, the infrared evidence is consistent with the "stable solubilizing adjuvant" target structure, proving that the target adjuvant has been successfully prepared.

[0143] Step three, preparation of ginkgo biloba flavone tablets

[0144] According to the weight part, 20 parts of L-arginine complex ginkgo biloba flavone prepared in Example 3, 28 parts of fast disintegrating solubilizing adjuvant, 45 parts of stable solubilizing adjuvant, 3 parts of cross-linked sodium carboxymethyl cellulose and 1 part of magnesium stearate were pressed into shape to obtain ginkgo biloba flavone tablets.

[0145] Comparative Example 1

[0146] The difference between this comparative example and Example 6 is that the L-arginine complex ginkgo biloba flavone used in the preparation process cancels the use of phenylboric acid solid silica gel adsorbent.

[0147] Comparative Example 2

[0148] The difference between this comparative example and Example 6 is that the L-arginine complex ginkgo biloba flavone used in the preparation process cancels step 4.

[0149] Comparative Example 3

[0150] The difference between this comparative example and Example 6 is that the stable solubilizing adjuvant is cancelled in step three.

[0151] Comparative Example 4

[0152] The difference between this comparative example and Example 6 is that the fast disintegrating solubilizing adjuvant is cancelled in step three.

[0153] Performance test:

[0154] The ginkgo biloba biflavones prepared in Examples 1-3 and Comparative Example 1 were determined for purity by high performance liquid chromatography. The sample obtained after purification was precisely weighed, dissolved in methanol and filtered through a 0.22 μm filter. The filtrate was injected into a C18 column, eluted with a gradient of acetonitrile and 0.1% phosphoric acid aqueous solution at a flow rate of 1.0 mL / min, and the column temperature was 35°C. The detection wavelength was 360 nm. The main biflavone components in the sample were determined by external standard method, and the peak area of each component was summed. The percentage of the peak area of the main components in the total peak area was taken as the purity of the sample. The purity of the ginkgo biloba biflavones was expressed as the percentage of the main components. The data are shown in Table 1.

[0155] Six tablets of the ginkgo biloba biflavones prepared in Examples 4-6 and Comparative Examples 1-4 were placed in the paddle apparatus of a dissolution tester. 900 mL of pH 6.8 phosphate buffer (containing 0.10% sodium dodecyl sulfate) was added as the dissolution medium. The water bath temperature was controlled at 37.0°C±0.5°C, and the paddle speed was 50 rpm. After 30 min, 10 mL of the sample was taken, filtered through a 0.45 μm filter, and determined for the concentration of the active ingredient by UV spectrophotometry at 360 nm. The dissolution rate of each tablet at 30 min was calculated, and the average dissolution rate was obtained.

[0156] Six tablets of the ginkgo biloba biflavones prepared in Examples 4-6 and Comparative Examples 1-4 were placed in the basket of a disintegration tester. 900 mL of pH 6.8 phosphate buffer was used as the medium, and the instrument was started at 37°C±0.5°C. The time required for complete disintegration of each tablet was recorded, and the average disintegration time was calculated.

[0157] Six tablets of the ginkgo biloba biflavones prepared in Examples 4-6 and Comparative Examples 1-4 were placed in a tablet hardness tester. The force required for tablet rupture was determined, and the average hardness of 10 tablets was recorded as the molding performance index.

[0158] The samples of the ginkgo biloba biflavones prepared in Examples 4-6 and Comparative Examples 1-4 were sealed and stored at 40°C±2°C / 75%±5%RH for 6 months. After storage, the samples were taken out, equilibrated at room temperature, and determined for dissolution rate, disintegration time, content uniformity, and hardness. The stability of the ginkgo biloba biflavones under high temperature and high humidity was evaluated. The storage time is shown in Table 2.

[0159] Table 1 - Performance test data of each sample

[0160]

[0161] Table 2 - Performance test data of each sample

[0162]

[0163] Data analysis:

[0164] After comparing and analyzing the data in Table 1-2, it is found that the ginkgo biflavonoid prepared has a purity of 98.5%, the ginkgo biflavonoid tablet prepared has an average dissolution rate of 95.9% in 30 minutes, an average disintegration time of 75s, and a tablet hardness of 86N, while the dissolution rate retention rate after aging is 96.7%, the disintegration rate retention rate is 94.6%, and the tablet hardness retention rate is 98.2%, all of which are better than the comparative examples, which shows that:

[0165] The comparative example 1 loses the selective complexation of the -B(OH)2 group in the phenylboronic acid solidified silica gel adsorbent to the ortho-diphenol structure of ginkgo biflavonoid, resulting in the residues of impurities such as flavonoids, polyphenols and sugars in the extraction liquid. The hydroxyl and carboxyl groups of the impurities are prone to hydrogen bonding with ginkgo biflavonoid, forming unstable multi-component complexes, which destroy the molecular dispersion state of flavonoids in the purified liquid. Such impurities promote particle agglomeration and interface hydrophobic shielding during drying and tabletting, hinder the water permeation channel of the fast-disintegrating and solubility-enhancing adjuvant, causing disintegration delay and dissolution limitation, and exacerbating oxidative degradation at elevated storage temperature;

[0166] The comparative example 2 loses the hydrogen bonding, electrostatic and π-π synergistic effect of the guanidino and amino groups in the L-arginine molecule to the phenolic hydroxyl and carbonyl groups of ginkgo biflavonoid, resulting in the re-aggregation of ginkgo biflavonoid in the purified liquid and the formation of local crystalline structure. During the spray drying process, the self-aggregates are prone to produce dense particles due to π-π stacking between molecules, reducing their specific surface area and hydrophilicity, resulting in insufficient structure porosity after tabletting, and a significant decrease in water permeation rate. At the same time, the absence of L-arginine leads to a decrease in the buffering and antioxidant capacity of the system, and the flavonoids undergo oxidative polymerization and color deepening at high temperature storage, resulting in a decrease in overall dissolution performance and stability;

[0167] The comparative example 3 loses the multi-phase support network constructed by β-cyclodextrin, microcrystalline cellulose and pregelatinized starch, so that the porous framework formed in the fast-disintegrating and solubility-enhancing adjuvant lacks morphological constraint and carrier support. After the spray coating film layer breaks, the particles can quickly disintegrate when encountering liquid, but the pore walls collapse and the particles agglomerate, resulting in unstable liquid passage and lack of skeleton regulation. The drug migrates unevenly in the system, and the dissolution behavior shows the characteristics of fast disintegration and fast sinking. When the temperature increases, the water migration and pore wall collapse are intensified, and the mechanical strength of the tablet and the reproducibility of drug release are significantly reduced;

[0168] Comparative Example 4 lost the porous rapid disintegration skeleton formed by mannitol, hydroxypropyl cellulose and arginine bicarbonate and the ethyl cellulose-tartaric acid composite film structure, so that the tablet system lacked the rapid water absorption swelling and liquid penetration pore-forming mechanism. The stable solubilizing adjuvant only showed slow surface swelling after encountering liquid, and the internal part was difficult to be rapidly wetted. The liquid penetration was blocked, leading to a surface erosion type disintegration. With the increase of storage temperature, the microcrystalline cellulose and citric acid film layer was further densified, the liquid penetration resistance was increased, the drug release was controlled by diffusion and was delayed, and the overall dissolution rate was significantly reduced;

[0169] Finally, it is pointed out that the process realizes the organic connection of structure and function in the links of raw material extraction, purification, compounding and preparation construction. The phenylboronic acid solidified silica gel adsorbent gives the system selective recognition and mild desorption characteristics in the pretreatment stage, providing high-purity raw materials for subsequent compounding. The introduction of L-arginine forms a stable dispersed structure of ginkgo biloba biflavones at the molecular level, significantly improving the film uniformity and composition stability during the drying process. The rapid disintegration solubilizing adjuvant establishes a rapid penetration channel through the porous skeleton and the cleavable film layer, while the stable solubilizing adjuvant maintains the tablet structure and thermal stability through cyclodextrin inclusion and cellulose support network. The functions of each part are closely linked to form a comprehensive system with rapid drug release, good mechanical strength and excellent storage stability. The prepared ginkgo biloba biflavone tablets are significantly superior to conventional preparation paths in terms of purity, dissolution and long-term retention performance.

[0170] The preferred embodiments disclosed above are only used to help explain the present application. The preferred embodiments do not describe all the details, nor limit the application to the specific embodiments. Obviously, many modifications and variations can be made according to the content of the present application. The present application is selected and specifically described to better explain the principles and practical applications of the present application, so that those skilled in the art can well understand and utilize the present application. The present application is limited by the claims and their entire scope and equivalents.

Claims

1. A product of ginkgetin characterized in that, The ginkgo biloba flavone product is obtained by compounding ginkgo biloba flavone and L-arginine, and the preparation method is as follows: L-arginine, ginkgo biloba flavone and 50wt% ethanol aqueous solution are added into a reaction kettle, stirred at room temperature for 15-20min, and then post-treated to obtain L-arginine-compounded ginkgo biloba flavone, wherein the dosage ratio of ginkgo biloba flavone, L-arginine and 50wt% ethanol aqueous solution is 3-4g:0.1g:30mL.

2. A product of ginkgetin according to claim 1, characterized in that, The extraction method of the ginkgo biloba flavone comprises the following steps: S1, take D 50 Ginkgo biloba leaf powder and deep eutectic solvent with a particle size of 100 μm are placed in an ultrasonic instrument, deionized water and pectinase and cellulase are added, the ultrasonic instrument is heated to 40-50℃, and ultrasonic extraction is carried out for 30 min, and then a ginkgo biloba biflavonoid dispersion liquid is obtained by post-treatment, wherein the amount ratio of the ginkgo biloba leaf powder, the deep eutectic solvent, the deionized water, the pectinase and the cellulase is 10g:15-20g:2-3mL:0.05g:0.03g, and the deep eutectic solvent is obtained by mixing choline chloride and lactic acid at an amount ratio of 1g:2g. S2, the ginkgo biloba flavone dispersion is added into a reaction kettle, and then saturated sodium bicarbonate aqueous solution is used to adjust the pH value of the reaction system to 8.5, after which phenyl boronic acid solidified silica gel adsorbent is added, stirred at room temperature for 30min, and then carbon dioxide gas is introduced, and post-treatment is performed to obtain ginkgo biloba flavone, wherein the dosage ratio of the ginkgo biloba flavone dispersion and the phenyl boronic acid solidified silica gel adsorbent is 25-30g:1g; The preparation method of the phenyl boronic acid solidified silica gel adsorbent is as follows: anhydrous ethanol, 3-aminophenyl boronic acid, triethylamine and isocyanic acid propyl triethoxy silane are added into a reaction kettle, stirred at room temperature for 30-40min, then silica gel, deionized water and glacial acetic acid are added into the reaction kettle, the temperature of the reaction kettle is increased to reflux, and the reaction kettle is kept at reflux for 3-4h, and post-treatment is performed to obtain the phenyl boronic acid solidified silica gel adsorbent, wherein the dosage ratio of anhydrous ethanol, 3-aminophenyl boronic acid, triethylamine, isocyanic acid propyl triethoxy silane, silica gel, deionized water and glacial acetic acid is 50-60mL:1g:0.2mL:3-4g:10g:0.1mL:0.02mL, the pore size of the silica gel is 60-80Å, and the particle size is 40-60μm.

3. Use of a ginkgetin product, characterized in that, The ginkgo biloba flavone product of claim 1 is applied to the preparation of ginkgo biloba flavone tablets, and the ginkgo biloba flavone tablets comprise the following raw materials in parts by weight: 20 parts of L-arginine-compounded ginkgo biloba flavone, 25-30 parts of rapid disintegration and solubilization auxiliary material, 40-50 parts of stable solubilization auxiliary material, 2-4 parts of crosslinked sodium carboxymethyl cellulose and 1 part of magnesium stearate; The preparation method of the rapid disintegration and solubilization auxiliary material comprises the following steps: A1, mannitol, hydroxypropyl cellulose, arginine bicarbonate and deionized water are added into a reaction kettle and stirred, and then spray drying is performed under the conditions of inlet air temperature 150℃, outlet air temperature 75℃, feeding rate 3-5mL / min and atomization gas pressure 0.5-0.7MPa to obtain porous framework particles; A2, tartaric acid, ethyl cellulose and anhydrous ethanol are added into a reaction kettle and stirred to prepare a spraying liquid, the spraying liquid is uniformly sprayed on the surface of the porous framework particles by using a fluidized bed, the film is made to have a weight gain of 8-10%, and post-treatment is performed to obtain the rapid disintegration and solubilization auxiliary material; The preparation method of the stable solubilization auxiliary material comprises the following steps: B1, corn starch and deionized water are added into a reaction kettle and stirred, and then the temperature of the reaction kettle is increased to 70-80℃, and the reaction kettle is kept at this temperature for 5-10min, and post-treatment is performed to obtain pregelatinized starch; B2, β-cyclodextrin, microcrystalline cellulose and pregelatinized starch were added into the reaction kettle and stirred uniformly, then anhydrous ethanol and deionized water were added for wet granulation, after granulation, D 50 =300µm particles were obtained, and the solubilization adjuvant precursor was obtained after post-processing; B3, under the condition of 45℃, the 0.5wt% citric acid aqueous solution is sprayed on the surface of the solubilization adjuvant precursor by fluidized bed, the film is increased by 4-5%, and then the stable solubilization adjuvant is obtained by post-processing.

4. The use of a product of ginkgetin according to claim 3, characterized in that, In step A1, the amount ratio of the mannitol, hydroxypropyl cellulose, arginine bicarbonate and deionized water is 7-8g:1g:1g:20mL; in step A2, the amount ratio of the tartaric acid, ethyl cellulose and anhydrous ethanol is 8-10g:3g:200mL.

5. The use of a product of ginkgetin according to claim 3, characterized in that, In step B1, the amount ratio of the corn starch and deionized water is 1g:3-4g; in step B2, the amount ratio of the β-cyclodextrin, microcrystalline cellulose, pregelatinized starch, anhydrous ethanol and deionized water is 5g:12-15g:1g:10mL:30mL.

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