Ryochito water extract, Ryochito water extract ointment, Ryochito preparations and their manufacturing method, and quality control standards for Ryochito preparations

Abstract

The present invention provides aqueous extracts of the two rehearsal decoctions, aqueous extract ointments of the two rehearsal decoctions, preparations of the two rehearsal decoctions, and methods for detecting the two rehearsal decoctions, whose raw materials are Rehmannia glutinosa, Xanthan Ginseng, Peony Root, Magnolia officinalis, Dioscorea chinensis, and Eucalyptus pith. The preparation method employs a concentration method in which the extracts are concentrated three times, which reduces the time required for concentration of low concentrations of paeoniflorin, shortens the absolute heating concentration time of the medicinal solution, and solves the problem of destruction of paeoniflorin, the active ingredient in the two rehearsal decoctions. The concentration after extraction is carried out at a relatively low temperature and under vacuum conditions, which helps protect and preserve the ingredients and avoids the destruction of ingredients such as paeoniflorin due to high temperatures.

Description

[Technical Field]

[0001] This application claims priority from a Chinese patent application filed with the China Patent Office on September 26, 2022, with application number CN202211173779.8, entitled "Liangji Tang aqueous extract, Liangji Tang aqueous extract ointment, Liangji Tang preparation and its manufacturing method, and quality control standards for Liangji Tang preparation," the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to the field of medicine, specifically to a water extract of two geotangs, a water extract ointment of two geotangs, a two geotang preparation and its manufacturing method, as well as quality control standards for two geotang preparations. [Background technology]

[0003] Liangdi Tang is derived from prescription number 90 in the Catalogue of Famous Formulas in Ancient Classics (1st batch) published by the State Administration of Traditional Chinese Medicine, and is originally from the Fu Qing Primary Nursing Medicine (Fushan, Qing Dynasty). It is composed of six herbal medicines: Rehmannia Root, Xuanshepherd's Ginseng, Peony Root, Wheat Root, Dijupi, and Ejiao. It is a classic prescription commonly used in gynecological menstrual regulation, with its main effects of nourishing yin, clearing heat, cooling blood, and regulating menstruation. It is mainly used to treat symptoms such as early menstruation, insufficient menstruation, prolonged menstruation, and dysfunctional uterine bleeding caused by yin deficiency and internal heat.

[0004] Patent Document 1 discloses a method for detecting the HPLC characteristic spectrum of Liangji Tang. The prior art also discloses a method for producing Liangji Tang using paeoniflorin and harpagoside as control substances, which involves adding water to five ingredients: 37.3g of Rehmannia glutinosa, 37.3g of Xanthan Ginseng, 18.6g of Peony Root, 18.6g of Wheat Flour, and 11.2g of Dioscorea Bark. The mixture is steeped for 40 minutes, then covered and extracted twice: the first time with 7 times the amount of water and extracted for 30 minutes, and the second time with 5 times the amount of water and extracted for 20 minutes. The decoction is filtered, the filtrates are combined and concentrated to approximately 500mL, and 11.2g of Erjiao is added and dissolved to obtain a standard decoction. The standard decoction is concentrated to approximately 250mL at low temperature and reduced pressure, and freeze-dried to obtain a freeze-dried powder. However, in the manufacturing method of the two local decoctions, the filtrates are combined and then concentrated, which takes time, and the heat-sensitive active ingredients in the two local decoctions, such as paeoniflorin, are easily decomposed, which affects the medicinal efficacy of the two local decoctions. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] China patent CN110412155A Summary of the Invention [Problem to be solved by the invention]

[0006] In view of this, the present invention aims to provide a water extract of the two geotangs, a water extract ointment of the two geotangs, a two geotang preparation, a method for producing the same, and quality control standards for the two geotang preparations. The water extract of the two geotangs, a water extract ointment, and a two geotang preparation provided by the present invention have a high content of heat-sensitive active ingredients such as paeoniflorin and have good medicinal effects. [Means for solving the problem]

[0007] To achieve the above object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a method for producing a water extract of Liangjiang, which includes the following steps: Five kinds of Chinese herbal medicines are mixed with first water, and then first soaking and first extraction are carried out in order to obtain a first extract and a first medicinal residue, and the five kinds of Chinese herbal medicines are Rehmannia Root, Xanthan Ginseng, Peony Root, Wheat Flour, and Dioscorea Root Pi. The first drug residue is mixed with second water, and a second soaking and a second extraction are carried out in sequence to obtain a second extract and a second drug residue. The second drug residue is mixed with third water, and a third soaking and a third extraction are carried out in sequence to obtain a third extract. The first extract, the second extract, and the third extract are each concentrated to obtain a first concentrate, a second concentrate, and a third concentrate, respectively, at a concentration temperature of 70 to 90°C, a vacuum degree of 0.03 to 0.07 MPa, and a steam pressure of ≦0.09 MPa. The first, second, and third concentrated solutions and the gelatin solution are mixed to obtain the aqueous extracts of both the tea decoctions. The mass ratio of the Rehmannia glutinosa root, Xanthan Ginseng root, Peony root, Euonymus rotundus, Dioscorea chinensis root and Euglena jiao was 37.30:37.30:18.65:18.65:11.19:11.19. Preferably, the mass of the first water is 6 to 8 times the mass of the five kinds of herbal medicines. The mass of the second water and the third water is independently 4 to 6 times the mass of the five kinds of Chinese herbal medicines. The temperatures of the first extraction, second extraction, and third extraction are independently 98 to 101°C. The first steeping and first extraction times are independently 30 to 50 minutes. The second steeping, second extraction, third steeping and third extraction times are independently 20 to 40 minutes. Preferably, the densities of the first, second and third concentrates are independently greater than or equal to 1.10 g / mL and less than 1.2 g / mL. Preferably, the method for preparing the gelatinous solution includes dissolving the gelatinous solution in water and then sieving the solution, the mass of the water being 2 to 6 times the mass of the gelatinous solution, and the sieve size of the sieve being 100 mesh.

[0009] The present invention provides a water extract of both geothermal tea and ginger obtained by the manufacturing method described in the above technical solution.

[0010] The present invention provides a double-ground water extract paste obtained by concentrating the double-ground water extract of the above technical solution, and the density of the double-ground water extract paste is 1.2-1.3 g / mL.

[0011] The present invention provides a dual-tang preparation, the raw materials of which include a dual-tang aqueous extract and a pharmaceutically acceptable excipient. The dual-tang aqueous extract is the dual-tang aqueous extract liquid described in the technical solution and / or the dual-tang aqueous extract ointment described in the technical solution. The mass ratio of the pharmaceutically acceptable excipient to the dual-tang aqueous extract is 1:(12.48-18.72), and the mass of the dual-tang aqueous extract is calculated based on the total mass of Rehmannia glutinosa, Xanthan Ginseng, Peony Root, Wheat Root, Dioscorea Root, and Egg Jiao.

[0012] Preferably, the raw materials for producing 1,000 parts of the Liangji Tang preparation include, in parts by mass, 468 to 572 parts of Rehmannia Root, 468 to 572 parts of Xanthan Ginseng, 234 to 286 parts of Peony Root, 234 to 286 parts of Wheat Flour, 140 to 171 parts of Dioscorea Rhizome, 140 to 171 parts of Erjiao, and 100 to 150 parts of a pharmaceutically acceptable excipient.

[0013] The present invention provides a method for producing the Liangjitang preparation described in the above technical solution, mixing an aqueous solution of the two geotang extract and a pharmaceutically acceptable excipient, and concentrating the mixture to obtain a concentrated mixture; and vacuum belt drying the mixed concentrate to obtain a Liangjiang preparation.

[0014] The present invention provides a quality control standard for the two geode preparations described in the above technical solution or the two geode preparations produced by the production method described in the above technical solution, including HPLC characteristic spectrum, characteristic component content, water content, and dry extraction rate of the two geode preparations.

[0015] The contents of the characteristic components are 0.33 to 0.67 mg / g of harpagoside, 2.62 to 4.96 mg / g of paeoniflorin, 8.83 to 16.42 mg / g of L-hydroxyproline, and 17.42 to 32.46 mg / g of glycine. The water content is 3 to 8 wt %. The dry extraction rate is 40 to 55%. [Effects of the Invention]

[0016] The present invention provides a method for producing a water extract of Liangji Tang. The post-extraction concentration is carried out at a relatively low temperature (70-90°C) and under vacuum (vacuum level 0.03-0.07 MPa, steam pressure ≤0.09 MPa), which helps protect and preserve the ingredients and avoids the destruction of heat-sensitive active ingredients (such as paeoniflorin) due to high temperatures. The traditional Chinese medicine is extracted multiple times, concentrated, combined, and then concentrated to a specified density. This method reduces the total concentration of the medicinal solution and extends the absolute heating and concentration time of the medicinal solution. At the same time, the present invention adopts a method of concentrating the extracts obtained by three extractions, so that the paeoniflorin content in the three extractions quickly reaches a high concentration, significantly shortening the low concentration concentration time of paeoniflorin (an environment in which paeoniflorin is easily destroyed) and the absolute heating concentration time of the medicinal liquid, solving the problem of destruction of paeoniflorin, the medicinal ingredient of both decoctions, and ensuring consistency between traditional decoctions and modern preparations. The obtained water extracts of both decoctions have a high content of effective active ingredients and excellent medicinal effects.

[0017] Modern traditional Chinese medicine only pursues extraction rate. The extraction time for a single extraction is usually 1-2 hours, and the excess extracted is often impurities such as fiber and starch, which does not increase the extraction of active ingredients. Prolonged heating can destroy the active ingredients. Compared with modern traditional Chinese medicine methods that require excessive extraction and high energy consumption, the production method provided by the present invention adds an appropriate amount of water during the extraction process, shortens the extraction time, and achieves high extraction efficiency (high extraction yields, paeoniflorin, and harpagoside), thereby further improving the quality and medicinal efficacy of the two-stage decoction product and reducing production costs.

[0018] Because the Liangjiang Decoction contains heat-sensitive ingredients such as viscous gelatin, polysaccharides, and paeoniflorin, which are prone to aggregation, traditional high-temperature drying and spray drying methods tend to cause aggregation, destroying heat-sensitive active ingredients such as paeoniflorin. The manufacturing method for the Liangjiang Decoction preparation provided by the present invention uses vacuum belt drying, which has the advantages of low drying temperature, short drying time, minimal loss, and high efficiency. The resulting Liangjiang Decoction preparation does not aggregate during the drying process, and heat-sensitive ingredients such as paeoniflorin are not destroyed. The Liangjiang Decoction preparation has uniform distribution of each ingredient, a high content of active ingredients such as paeoniflorin, and good medicinal efficacy.

[0019] The present invention also provides quality control standards for the Liangjiang Tang preparations described in the above technical solutions or those prepared by the manufacturing methods described in the above technical solutions. The present invention employs multi-component indicators (paeoniflorin content, harpagoside content, water content, dry extract rate) plus characteristic spectrum technology to comprehensively monitor the overall chemical composition of the Liangjiang Tang preparations. The quality standards are more comprehensive, which is important for ensuring the stability and high quality of the Liangjiang Tang preparation products. [Brief explanation of the drawings]

[0020] [Figure 1] HPLC control characteristic spectra of the reference samples of both the decoctions. [Figure 2] HPLC contrast characteristic spectra of both Ji Tang preparations. [Figure 3] This is a diagram showing the water absorption rules for Ryochiyu drinking pieces. [Figure 4] FIG. 1 is a graph showing the elution curve of harpagoside. [Figure 5] FIG. 1 is a dissolution curve of paeoniflorin. [Figure 6] 1 is a liquid phase spectrum of paeoniflorin in series 1 test substance solutions. [Figure 7] 1 is a liquid phase spectrum of paeoniflorin in series 2 test substance solutions. [Figure 8] 1 is a liquid phase spectrum of paeoniflorin in series 3 test substance solutions. [Figure 9] 1 is a liquid phase spectrum of paeoniflorin in series 4 test substance solutions. [Figure 10] 1 is a liquid phase spectrum of paeoniflorin in series 5 test substance solutions. [Figure 11] Test results of paeoniflorin solutions of different concentrations, showing changes with heating time. [Figure 12] This is a chromatogram of the changes after heating a 0.05 mg / mL paeoniflorin solution for 4 hours. [Figure 13] This is a chromatogram of the changes after heating paeoniflorin solutions of different concentrations for 4 hours. DETAILED DESCRIPTION OF THE INVENTION

[0021] The present invention provides a method for producing a water extract of Liangjiang, which includes the following steps: Five kinds of herbal medicines are mixed with first water, and then first soaking and first extraction are carried out in order to obtain a first extract and a first medicinal residue. The five herbal medicines are Rehmannia Root, Xanthan Ginseng, Peony Root, Wheat Root, and Dioscorea Pelargonium. The first drug residue is mixed with second water, and a second soaking and a second extraction are carried out in sequence to obtain a second extract and a second drug residue. The second drug residue is mixed with third water, and a third soaking and a third extraction are carried out in sequence to obtain a third extract. The first extract, the second extract, and the third extract are each concentrated to obtain a first concentrate, a second concentrate, and a third concentrate, respectively, at a concentration temperature of 70 to 90°C, a vacuum degree of 0.03 to 0.06 MPa, and a steam pressure of 0.03 to 0.05 MPa, independently. The first, second, and third concentrated solutions and the gelatin solution are mixed to obtain the aqueous extracts of both the tea decoctions.

[0022] The mass ratio of the Rehmannia glutinosa root, Xanthan Ginseng root, Peony root, Euonymus rotundus, Dioscorea chinensis root and Euglena jiao was 37.30:37.30:18.65:18.65:11.19:11.19.

[0023] In the present invention, unless otherwise specified, all raw materials are commercially available products familiar to those skilled in the art.

[0024] In the present invention, five kinds of Chinese herbal medicines are mixed with first water, and first steeping and first extraction are carried out in order to obtain a first extract and a first medicinal residue, and the five kinds of Chinese herbal medicines are Rehmannia Root, Xuanshen Root, Peony Root, Ophiopogon Root, and Dijupi Root. In the present invention, the mass ratio of Rehmannia Root, Xuanshen Root, Peony Root, Ophiopogon Root, and Dijupi Root is 37.3:37.3:18.65:18.65:11.9.

[0025] In the present invention, the five herbal medicines are preferably sieved before use. In the present invention, the sieving of the Rehmannia glutinosa preferably includes the following steps: (1) Selection: Remove impurities; (2) Spray cleaning: 10-15 minutes; (3) Moisturizing: Moisturize at 15-35°C for 8-12 hours; (4) Cutting: Cut into 2-4mm thick slices with a 3mm gap between the blade and the cutting edge; (5) Drying: Dry at 60-70°C for 6-7 hours to a thickness of 2-3cm; (6) Wine roasting: Add 15% yellow wine and roast at 220°C for 24 minutes until the surface temperature of the medicine reaches 130-135°C; (7) Sieving: Sift the fragments through a 3mm medicine sieve, and manually select any burnt or unacceptable pieces. In the present invention, the sieving of the Xuan Sheng Xianthus root preferably includes the following steps in order: (1) Selection: Remove impurities. (2) Spray cleaning: 15-20 minutes. (3) Moisturizing: Moisturize at 5-35°C for 4-26 hours. (4) Cutting: Cut into 1-2 mm slices with a 1.5 mm gap between the blade and the cutting edge. (5) Drying: Dry to a thickness of 2-3 cm at a temperature of 50-60°C for 3.5-4.5 hours. (6) Selection: Sift the fragments through a 2 mm sieve, and manually select rejected pieces that are burnt or have an unacceptable thickness. In the present invention, the processing of the liquor-brewed white peony root preferably includes the following steps: (1) Selection: Remove impurities and classify (small class: root apical diameter < 1.0 cm, large class: root apical diameter ≥ 1.0 cm). (2) Spray cleaning: 10-15 minutes. (3) Infiltration: Infiltration at 15-35°C for 12-40 hours for small class and 24-56 hours for large class. (4) Cutting: The gap between the blade and the cutting edge is 1.5 mm, and the slices are cut into 1-2 mm thick slices. (5) Drying: The slices are dried at a temperature of 70-80°C for 4-5 hours at a thickness of 2-3 cm. (6) Wine-roasting: The slices are added with 20% yellow wine and roasted at 230°C for 20 minutes, until the surface temperature of the slices reaches 140-145°C. (7) Selection: The slices are sieved through a 3 mm sieve to remove debris, and rejected slices, such as those that are burnt or have an unacceptable thickness, are manually selected. In the present invention, the production of wheat bran pie preferably includes the following steps: (1) Selection: Removes impurities. (2) Spray cleaning: 10-15 minutes. (3) Moisturizing: Moisturizes at 5-35°C for 6-24 hours. (4) Flattening: The gap between the shafts is 2-3 mm, and the slices are compressed into flat pieces. (5) Drying: The slices are dried at a temperature of 50-60°C for 6-7 hours at a thickness of 2-3 cm.(6) Careful selection: Manually select unsatisfactory products that have not been flattened. In the present invention, the process of processing the bone peel preferably includes the following steps: (1) Careful selection: Remove impurities and residual wood core. (2) Cleaning: The standard is that the original color is displayed and there is no dust or impurities. (3) Drying: Dry at a temperature of 40-50°C for 4-5 hours to a thickness of 2-3cm. (4) Sift the fragments through a 3mm sieve.

[0026] In the present invention, the mass of the first water is preferably 6 to 8 times, more preferably 6 to 7.5 times, and even more preferably 6.5 to 7 times the mass of the five herbal medicines. In the present invention, the temperature of the first steeping is preferably room temperature, and the first steeping time is preferably 30 to 50 minutes, more preferably 35 to 40 minutes. In the present invention, the temperature of the first extraction is preferably 98 to 101°C, more preferably 99 to 100°C. The first extraction time is preferably 30 to 50 minutes, more preferably 35 to 40 minutes.

[0027] The present invention preferably further comprises post-treating the obtained first extraction system after completing the first extraction. The post-treatment preferably comprises centrifuging the obtained first extraction system to obtain a liquid component and a first drug residue, and microfiltration of the liquid component to obtain a first extract. In the present invention, the centrifugation is preferably performed using a centrifuge, and the conditions for the centrifugation are not particularly limited as long as the liquid component and solid component can be separated. In the present invention, the microfiltration is preferably performed using a plate and frame filter, and the pore size of the filter membrane of the plate and frame filter is preferably 5 to 15 μm.

[0028] In the present invention, after obtaining the first drug residue, the first drug residue is mixed with second water, and second soaking and second extraction are sequentially performed to obtain a second extract and a second drug residue. In the present invention, the mass of the second water is preferably 4 to 6 times the mass of the five herbal medicines, more preferably 4 to 5.5 times, and even more preferably 4.5 to 5 times. In the present invention, the temperature of the second soaking is preferably room temperature, and the temperature of the second extraction is preferably the same as the temperature of the first extraction, although this will not be described in detail here. The time periods for the second soaking and second extraction are independently preferably 20 to 40 minutes, more preferably 20 to 35 minutes, and even more preferably 25 to 30 minutes. In the present invention, after completing the second extraction, the obtained second extraction system is preferably post-treated, and this post-treatment is preferably the same as the post-treatment method for the first extraction system described above, although this will not be described in detail here.

[0029] In the present invention, after obtaining the second drug residue, the second drug residue is mixed with third water, and a third steeping and a third extraction are sequentially performed to obtain a third extract. In the present invention, the mass of the third water is preferably 4 to 5 times, more preferably 4.5 times, the mass of the five herbal medicines. In the present invention, the temperature of the third steeping is preferably room temperature, and the temperature of the third extraction is preferably the same as the temperature of the first extraction, although this will not be described in detail here. The time periods for the third steeping and the third extraction are independently preferably 20 to 40 minutes, more preferably 20 to 35 minutes, and even more preferably 25 to 30 minutes. In the present invention, after completing the third extraction, the obtained second extraction system is preferably post-treated. This post-treatment is the same as the post-treatment method for the first extraction system described above, but this will not be described in detail here.

[0030] In the present invention, after obtaining a first extract, a second extract, and a third extract, the first extract, the second extract, and the third extract are each concentrated to obtain a first concentrate, a second concentrate, and a third concentrate, respectively. In the present invention, the temperatures of the first concentrate, the second concentrate, and the third concentrate are independently 70 to 90°C, preferably 75 to 85°C, and more preferably 80°C. The degree of vacuum is independently 0.03 to 0.07 MPa, preferably 0.04 to 0.06 MPa. The steam pressure is independently 0.09 MPa or less, preferably 0.03 to 0.05 MPa. In the present invention, there are no particular limitations on the times of the first concentrate, the second concentrate, and the third concentrate, as long as the densities of the first concentrate, the second concentrate, and the third concentrate are independently concentrated to 1.10 g / mL or more and less than 1.2 g / mL.

[0031] In the present invention, after obtaining the first, second, and third concentrates, the first, second, and third concentrates are mixed with the gelatin solution to obtain a water extract of both Rehmannia glutinosa extracts. The mass ratio of the Rehmannia glutinosa root, Xanthan Ginseng root, Peony root, Wheat Root, and gelatin solution is 37.30:37.30:18.65:18.65:11.19:11.19. In the present invention, the method for producing the gelatin solution preferably includes the following steps: dissolving the gelatin solution in water and then sieving it, with the remaining residue being the gelatin solution. In the present invention, the mass of the water is preferably 2 to 6 times, more preferably 3 to 5 times, and even more preferably 4 times the mass of the gelatin solution. In the present invention, the dissolution is preferably carried out in a hot boiling water bath under stirring conditions. The stirring speed and time are not particularly limited in the present invention, as long as the gelatin solution is homogeneously dissolved. The present invention preferably further comprises removing bubbles and impurities from the upper layer of the dissolution system after dissolution, and then sieving the mixture. In the present invention, the sieve size is preferably 100 mesh. The present invention does not particularly limit the mixing method as long as the raw materials can be mixed uniformly.

[0032] The present invention provides a water extract of both geothermal tea and herbs obtained by the manufacturing method described in the above technical solution. In the present invention, the density of the water extract of both geothermal tea and herbs is preferably 1.2-1.3 g / mL, more preferably 1.25 g / mL.

[0033] The present invention provides a double-ground water extract paste obtained by concentrating the double-ground water extract described in the above technical solution. In the present invention, the density of the double-ground water extract paste is 1.2 to 1.3 g / mL, preferably 1.22 to 1.28 g / mL, and more preferably 1.25 g / mL. In the present invention, the concentration conditions are preferably the same as those in the process for producing the first concentrate, but will not be described in detail here. In the present invention, there is no particular limitation on the concentration time, as long as the concentrate is concentrated to a density of 1.2 to 1.3 g / mL.

[0034] The present invention provides a dual-tang preparation, the raw materials of which include a dual-tang aqueous extract and a pharmaceutically acceptable excipient. The dual-tang aqueous extract is the dual-tang aqueous extract liquid described in the technical solution and / or the dual-tang aqueous extract ointment described in the technical solution. The mass ratio of the pharmaceutically acceptable excipient to the dual-tang aqueous extract is 1:(12.48-18.72), preferably 1:(13-18), more preferably 1:(14-17), and even more preferably 1:(15-16). The mass of the dual-tang aqueous extract is calculated based on the total mass of Rehmannia glutinosa, Xanthan Ginseng, Peony Root, Wheat Root, Dioscorea Root, and Egg Gluconolactone. The pharmaceutically acceptable excipient preferably includes one or more of dextrin, starch, and lactose.

[0035] In the present invention, when calculated by mass fraction, the raw materials for producing 1,000 parts of a Ryochito preparation are preferably, expressed in parts by mass, 468 to 572 parts (more preferably 480 to 550 parts, even more preferably 500 to 520 parts) of Rehmannia glutinosa root, 468 to 572 parts (more preferably 480 to 550 parts, even more preferably 500 to 520 parts) of Xiangyang root, 234 to 286 parts (more preferably 240 to 270 parts, even more preferably 250 to 260 parts) of Liquor White Peony Root, 234 to 286 parts (more preferably 240 to 270 parts, even more preferably 250 to 260 parts) of Mugidzu ... Mugidzu root, 234 to 286 parts (more preferably 240 to 270 parts, even more preferably 250 to 260 parts) of Mugidzu root, 234 to 286 parts (more preferably 240 to 270 parts, even more preferably 250 to 260 parts) of Mugidzu root, 234 to 286 parts (more preferably 240 to 270 parts, even more preferably 250 to 260 parts) of Mugidzu root, 234 to 286 parts (more preferably Each of the two Jingde formulations contains 234 to 286 parts (more preferably 240 to 270 parts, even more preferably 250 to 260 parts), 140 to 171 parts (more preferably 150 to 170 parts, even more preferably 156 to 165 parts) of Jingde cortex, 140 to 171 parts (more preferably 150 to 170 parts, even more preferably 156 to 165 parts) of Erjiao, and 100 to 150 parts (more preferably 110 to 140 parts, even more preferably 120 to 130 parts) of a pharmaceutically acceptable excipient. In the present invention, the water content of the two Jingde formulations is preferably 3 to 8 wt%, more preferably 4 to 7 wt%, even more preferably 5 to 6 wt%. In the present invention, the dry extract ratio of the two Jingde formulations is preferably 40.0 to 55.0%, more preferably 45 to 50%.

[0036] In the present invention, the preparation preferably includes powder or granules of the two-way decoction. The particle size of the powder is preferably 150-850 μm, and the particle size of the granules is preferably 180-2000 μm.

[0037] The present invention provides a method for producing the Liangji Tang preparation described in the above technical solution, which includes the steps of mixing the aqueous extract of Liangji Tang with an aqueous solution of pharmaceutically acceptable excipients, concentrating the resulting mixed concentrate, and vacuum belt drying it to obtain the Liangji Tang preparation (i.e., Liangji Tang powder).

[0038] In the present invention, the aqueous solution of the pharmaceutically acceptable excipient is preferably obtained by dissolving the pharmaceutically acceptable excipient in water. The mass ratio of the pharmaceutically acceptable excipient to water is preferably 1:(2-6), more preferably 1:(3-5), and even more preferably 1:4. The dissolution is preferably carried out under stirring conditions. In the present invention, there are no particular limitations on the speed or time of the stirring, as long as a uniform aqueous solution of the pharmaceutically acceptable excipient is obtained.

[0039] In the present invention, the mixing and concentration conditions are preferably the same as the concentration conditions in the production process of the first concentrate, but will not be described in detail here. In the present invention, there are no particular limitations on the mixing and concentration time, as long as the mixture is concentrated until the resulting mixture concentrate density is 1.25 to 1.36 g / mL. The density of the mixture concentrate is more preferably 1.28 to 1.34 g / mL, and even more preferably 1.30 to 1.32 g / mL.

[0040] In the present invention, the vacuum belt drying is preferably performed using a vacuum belt dryer. The parameters of the vacuum belt drying include the following: The supply temperature is preferably 75 to 85°C, more preferably 80°C. The supply rate is preferably 6 to 24 L / h, more preferably 10 to 20 L / h. The crawler belt running speed is preferably 16±5 cm / min, more preferably 16±2 cm / min. The swing arm angle is preferably 65 to 85°C, more preferably 70 to 80°C. The fabric motor speed is preferably 30 to 50 r / min, more preferably 40 r / min. The heating temperatures of zones 1, 2, 3, and 4 are preferably 95±5°C (more preferably 95±2°C), 96±5°C (more preferably 96±2°C), 96±5°C (more preferably 96±2°C), and 30±5°C (more preferably 30±2°C, cooling), respectively. The cutter speed is preferably 10 to 20 seconds per cycle, more preferably 15 seconds per cycle. The degree of vacuum is preferably 0.097 to 0.1 MPa, and more preferably 0.098 to 0.099 MPa.

[0041] In the present invention, when the double-decoction preparation is a double-decoction granule, the method preferably further comprises granulating the double-decoction granule after the vacuum belt drying to obtain the double-decoction granule. In the present invention, the granulation is preferably carried out using a granulator. The granulation parameters include the following: the roller pressure of the granulator is preferably 12 to 20 MPa, more preferably 15 to 18 MPa; the vertical rotation speed of the screw feeder is preferably 18 to 30 r / min, more preferably 20 to 25 r / min, and the horizontal rotation speed of the screw feeder is preferably 90 to 140 r / min, more preferably 100 to 120 r / min; the rotation speed of the roller is preferably 4 to 10 r / min, more preferably 5 to 8 r / min; and the sizing speed is preferably 100 to 150 r / min, more preferably 120 to 130 r / min. In a specific embodiment of the present invention, during the granulation process, it is preferable to observe the status of the granulator (whether the powder supply of the equipment is smooth, whether the operating pressure is stable, whether the granules are discharged smoothly, etc.) and the properties of the granules (whether the granules are uniform, etc.), and according to the operating status of the equipment and the properties of the granules, adjust the equipment parameters (roller pressure, feeding speed, etc.), add powder in a timely manner, collect the granules, record the adjustment status of the granulation parameters, turn on the vibrating screen, put the granules after granulation into the hopper, adjust the hopper valve so that the feeding speed can completely separate the qualified granules (particle size of 180-2000 μm) from the fine powder, and put the qualified granules (Liangjitang granules) into the transfer container barrel, and re-granulate the fine powder.

[0042] The present invention also provides quality control standards for the two geode preparations described in the above technical solution or the two geode preparations prepared by the preparation methods described in the above technical solution, including HPLC characteristic spectra, characteristic component contents, water contents and dry extraction rates.

[0043] The contents of the characteristic components are 0.33 to 0.67 mg / g of harpagoside, 2.62 to 4.96 mg / g of paeoniflorin, 8.83 to 16.42 mg / g of L-hydroxyproline, and 17.42 to 32.46 mg / g of glycine. The water content is 3 to 8 wt %. The dry extraction rate is 40 to 55%.

[0044] In the present invention, the harpagoside is an indicator of the mass content of Xanthan Ginseng, the paeoniflorin is an indicator of the mass content of Peony Root, and the L-hydroxyproline and glycine are indicators of the mass content of Erwinia Root.

[0045] In the present invention, the method for obtaining the HPLC characteristic spectrum of the above-mentioned two rendang preparations preferably includes the following steps: (1) The test subject Ryojito preparation, diatomaceous earth and acetonitrile aqueous solution are mixed and concentrated, and the resulting concentrate is dissolved in a methanol aqueous solution to obtain a test substance solution. (2) Mix the paeoniflorin control substance, the harpagoside control substance, and methanol to obtain a control substance solution. (3) The test substance solution and the control substance solution are subjected to HPLC detection, respectively, to obtain the control substance characteristic spectrum and the test substance characteristic spectrum. Using the retention time of the paeoniflorin control substance in the control substance characteristic spectrum as a reference, the relative retention times of each of the other characteristic peaks in the test substance characteristic spectrum are calculated to obtain the HPLC characteristic spectra of both the Jidang preparations. Step (1) and Step (2) have no chronological order.

[0046] In the present invention, the test substance solution is obtained by mixing the test subject Ryoji Tang preparation, diatomaceous earth, and an acetonitrile aqueous solution, followed by concentration, and dissolving the resulting concentrate in a methanol aqueous solution. In the present invention, the concentration of the test subject Ryoji Tang preparation in the test substance solution is preferably 0.03 to 0.10 g / mL, more preferably 0.05 to 0.08 g / mL. In the present invention, the mass ratio of the Ryoji Tang preparation to diatomaceous earth is preferably 1:1 (0.5 to 1.5), more preferably 1:1. In the present invention, the volume fraction of acetonitrile in the acetonitrile aqueous solution is preferably 70 to 90%, more preferably 80%. In the present invention, the volume fraction of methanol in the methanol aqueous solution is preferably 5 to 15%, more preferably 10%. In a specific embodiment of the present invention, the method for preparing the test substance solution preferably includes the following steps: The two Jitang preparations to be tested were finely pulverized, and 1 g of powder of the two Jitang preparations to be tested was accurately weighed and placed in a stoppered Erlenmeyer flask. 1 g of diatomaceous earth was added and mixed uniformly, and 25 mL of 80% acetonitrile aqueous solution (acetonitrile volume fraction 80%) was accurately added, weighed, and ultrasonicated at room temperature, 800 W, 40 kHz for 20 minutes. After cooling, the mixture was weighed again, and the lost weight was compensated for using 80% acetonitrile aqueous solution. Shaking uniformly and filtering were performed, and 15 mL of the filtrate was accurately measured and evaporated to dryness. The residue was dissolved in 10% methanol aqueous solution, transferred to a 10 mL measuring flask, and the volume was adjusted to the specified volume with 10% methanol aqueous solution. Shaking uniformly, the mixture was filtered through a microporous membrane, and the resulting filtrate was used as the test substance solution.

[0047] In the present invention, a control substance solution is obtained by mixing a paeoniflorin control substance, a harpagoside control substance, and methanol. In the present invention, the concentration of the paeoniflorin control substance in the control substance solution is preferably 180 to 200 μg / mL, and the concentration of harpagoside in the control substance solution is preferably 10 to 30 μg / mL, more preferably 20 μg / mL.

[0048] In the present invention, after obtaining the test substance solution and the control substance solution, HPLC detection is performed on the test substance solution and the control substance solution respectively to generate reference characteristic spectra, and based on the retention time of the chromatographic peak of the control substance, the relative retention times of other common peaks are calculated to obtain the HPLC characteristic spectra of the two Reitang preparations.

[0049] In the present invention, the HPLC detection conditions include the following: a chromatography column using octadecylsilane-bonded silica as a packing material; mobile phase A is methanol, mobile phase B is 0.05 v / v% phosphoric acid solution, and the mobile phase flow rate is 0.8 to 1.2 mL / min, preferably 1 mL / min; the elution method is gradient elution, and the gradient elution procedure is shown in Table 1; the detection wavelength is 254 nm; the column temperature is 28°C, and the sample load volume is 10 μL; and the theoretical plate number should be 2000 or more based on the paeoniflorin peak.

[0050] JPEG2025532254000001.jpg50170

[0051] In the present invention, as shown in Figure 2, the characteristic spectrum of the test substance shows 10 characteristic peaks, of which two peaks should match the peak retention times of the corresponding reference substances (paeoniflorin reference substance and harpagoside reference substance), and the peak corresponding to the paeoniflorin reference substance peak is the S peak. The relative retention times of each characteristic peak and the S peak are calculated, and the relative retention times should be within ±10% of the specified value, and the specified value is preferably Preferably, the peaks are 0.22 (peak 1), 0.26 (peak 2), 0.31 (peak 3), 0.33 (peak 4), 1.00 (peak 5), 1.47 (peak 6), 1.54 (peak 7), 1.82 (peak 8), 1.91 (peak 9), and 2.22 (peak 10), of which peaks 1 to 3 are unknown components, peak 4 is gallic acid, peak 5 is paeoniflorin, peak 7 is verbascoside, peak 8 is angoloside C, peak 9 is cinnamic acid, and peak 10 is harpagoside.

[0052] In the present invention, the dry extraction rate is preferably tested with reference to General Rule 0104 of the 2020 edition of the Chinese Pharmacopoeia.

[0053] The two test preparations were finely pulverized, and 4 g of the powder was added to 30 mL of methanol, sonicated for 30 minutes, filtered, and the filtrate was concentrated to 5 mL for use as the test substance solution. As a control, 1.5 g of dijupi was added to 30 mL of methanol, sonicated for 30 minutes, filtered, and the filtrate was concentrated to 5 mL for use as the control solution. 10 μL of each of the test substance solution and control solution was spotted onto the same silica gel G thin-layer plate, using cyclohexane-ethyl acetate-formic acid (volume ratio 5:5:0.4) as the developing solvent. After drying, the plate was examined under ultraviolet light (365 nm). In the chromatogram of the test substance, the fluorescent spot of the same color appearing at the corresponding position on the chromatogram of the control was dijupi.

[0054] In the present invention, the method for testing the moisture content is not particularly limited, and any method for testing the moisture content that is well known to those skilled in the art, such as a drying method, can be used.

[0055] In the present invention, the testing method for the content of paeoniflorin and harpagoside is the same as the method for obtaining the HPLC characteristic spectra of the above-mentioned two decoction preparations, so it will not be further described here.

[0056] In the present invention, the method for testing the content of L-hydroxyproline and glycine preferably includes the following steps: (1) Finely crush the test preparations, place 1.5 g of the powder in a 25 mL measuring flask, add 20 mL of 0.1 mol / L hydrochloric acid, and sonicate for 30 minutes. After cooling, add 0.1 mol / L hydrochloric acid up to the line and shake evenly to obtain a hydrochloric acid solution of the test preparations. Place 2 mL of the hydrochloric acid solution of the test preparations in a 10 mL ampoule, add 2 mL of 0.08-0.12 mol / L hydrochloric acid, and hydrolyze at 150°C for 1 hour. Cool, transfer to an evaporating dish, wash with 10 mL of water in batches, and evaporate the washings into the evaporating dish to dryness. Dissolve the residue in 0.1 mol / L hydrochloric acid, transfer to a 25 mL measuring flask, add 0.1 mol / L hydrochloric acid up to the line, and shake evenly to obtain a test substance solution. (2) Mix the L-hydroxyproline control substance, the glycine control substance, and an aqueous hydrochloric acid solution to obtain a control substance solution, in which the concentration of L-hydroxyproline in the control substance solution is 60 μg / mL and the concentration of glycine in the control substance solution is 130 μg / mL. (3) Accurately measure 1 mL of the test substance solution into a 25 mL stoppered test tube, add 0.5 mL of 0.1 mol / L phenylisothiocyanate (PITC) in acetonitrile and 0.5 mL of 1 mol / L triethylamine in acetonitrile, shake evenly, and leave at room temperature for 1 hour. Then add 3 mL of 50% (v / v) aqueous acetonitrile solution, shake evenly, add 5 mL of n-hexane, shake, and leave for 10 minutes. Remove the lower layer of the solution and filter it through a microporous membrane. The resulting filtrate is the derivatized test substance solution. (4) The test substance solution of step (3) is replaced with the control substance solution to obtain a derivatized control substance solution. (5) The derivatized test substance solution and the derivatized control substance solution are subjected to HPLC detection to obtain the contents of L-hydroxyproline and glycine. Steps (1) and (2) have no chronological order. Steps (3) and (4) have no chronological order.

[0057] In the present invention, the detection conditions for the HPLC detection preferably include the following: a chromatography column using octadecylsilane-bonded silica as a packing material is used; acetonitrile-0.1 mol / L sodium acetate solution (adjusted to pH 6.5 with acetic acid) (7:93) is used as mobile phase A; acetonitrile-water (volume ratio 4:1) is used as mobile phase B; and the elution method is gradient elution, the gradient elution procedure of which is shown in Table 2. The mobile phase flow rate is 0.8 mL / min. The detection wavelength is 254 nm. The column temperature is 43°C, and the sample load amount is 5 μL. The theoretical plate number is 4,000 to 10,000, calculated based on the L-hydroxyproline peak.

[0058] JPEG2025532254000002.jpg59170

[0059] The following will clearly and completely describe the technical solutions of the present invention with reference to the embodiments of the present invention. Obviously, the described embodiments are only some of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, any other embodiments obtained by those skilled in the art without any creative efforts fall within the scope of protection of the present invention.

[0060] In the following examples of the present invention, each herbal medicine is prepared by pouring prior to use. According to the pouring requirements of each herbal ingredient contained in Liangjiang Tang in pharmacological research, the pouring techniques are studied through testing, and detailed pouring technique parameters such as soaking, cutting, drying, and the amount of excipients added are determined. The pouring method for each herbal medicine is as follows: Wine Rehmannia: (1) Selection: Remove impurities. (2) Spray cleaning: 10-15 minutes. (3) Moisturizing: Moisturize at 15-35°C for 8-12 hours. (4) Cutting: Cut into 2-4mm thick slices with a 3mm gap between the blade and the tip. (5) Drying: Dry at 60-70°C for 6-7 hours to a thickness of 2-3cm. (6) Wine-roasting: Add 15% wine and fry at 220°C for 24 minutes until the surface temperature of the rehmannia reaches 130-135°C. (7) Careful selection: Sift the fragments through a 3mm sieve, and manually select any burnt or unacceptable pieces. Xuanjin: (1) Selection: Remove impurities. (2) Spray cleaning: 15-20 minutes. (3) Moisturizing: Moisturize at 5-35°C for 4-26 hours. (4) Cutting: Cut into 1-2 mm slices with a 1.5 mm gap between the blade and the cutting edge. (5) Drying: Dry to a thickness of 2-3 cm at a temperature of 50-60°C for 3.5-4.5 hours. (6) Careful selection: Sift the fragments through a 2 mm sieve, and manually select any burnt or unacceptable pieces. Rice wine white peony: (1) Selection: Remove impurities and classify (small class: root apical diameter <1.0cm, large class: root apical diameter ≥1.0cm). (2) Spray washing: 10-15 minutes. (3) Infiltration: 15-35℃, 12-40 hours for small class, 24-56 hours for large class. (4) Cutting: Cut into 1-2mm thick slices with a 1.5mm gap between the blade and the tip. (5) Drying: Dry at 70-80℃ for 4-5 hours to a thickness of 2-3cm. (6) Wine roasting: Add 20% yellow wine and roast at 230℃ for 20 minutes until the surface temperature of the peony reaches 140-145℃. (7) Careful selection: Sift the fragments through a 3mm sieve, and manually select any burnt or unacceptable pieces. Barley winter: (1) Selection: Remove impurities. (2) Spray cleaning: 10-15 minutes. (3) Moisturizing: Moisturize at 5-35°C for 6-24 hours. (4) Flattening: Compress into a flat section with a 2-3mm shaft gap. (5) Drying: Dry to a thickness of 2-3cm at a temperature of 50-60°C for 6-7 hours. (6) Careful selection: Manually select unqualified products that have not been flattened. Jigokupi: (1) Selection: Remove impurities and residual wood core. (2) Cleaning: The standard is to show the original color and be free of dust and impurities. (3) Drying: Dry for 4-5 hours at a temperature of 40-50°C to a thickness of 2-3cm. (4) Sift through a 3mm sieve to remove debris.

[0061] In the following examples, the preparation method of the accompanying decoction (reference sample of the same batch) is as follows: Five herbal medicines, namely, Rehmannia glutinosa 37.3g, Xanthan Ginseng 37.3g, Peony Root 18.6g, Wheat Flour 18.6g, and Dijupi Root 11.2g, are taken, added with water and steeped for 40 minutes, added with water 7 times the weight of the five herbal medicines, boiled for 30 minutes, and filtered to obtain a first extract and a first herbal medicine residue, added with water 5 times the weight of the five herbal medicines to the obtained first herbal medicine residue, boiled for 20 minutes, and filtered to obtain a second extract, combined with the first extract and the second extract, concentrated to about 500mL, added with 11.2g of Ejiao to dissolve, concentrated under reduced pressure to about 250mL, and freeze-dried to a constant weight, and the resulting freeze-dried powder is the accompanying decoction (reference sample of the same batch).

[0062] Example 1 Research on prescription and dosage of Ryojito Liangchi Tang originates from the Fu Qingzhu Nuke, written by Fu Shan (also known as Qingzhu), a physician in the late Ming and early Qing dynasties. The prescription (daily dosage) listed in the original text was: dadiji (1 liang, stir-fried with wine), yuanseng (1 liang), white peony (5 sen, stir-fried with wine), maize winter radish (5 sen), digupi (3 sen), and ejiao (3 sen).

[0063] Pharmacognosy research was conducted on each herb, and since the prescription was written by Fu Shan, a physician from the late Ming and early Qing dynasties, this study mainly studied the relevant descriptions of each herb in pharmacological literature from the Ming and Qing dynasties onwards, and compared them with the 2020 editions of the Chinese Pharmacopoeia, Chinese Materia Medica and related literature to determine the original plant variety and preparation method of each herb. Da dǎo ​​(stir-fried with wine) is Rehmannia (roasted with wine), Yuan shang (sage) is Xanthan Ginseng, White Peony (stir-fried with wine) is White Peony in Wine, Wheat Dong Rou (wheat dong), Di Bù Pi (di bone skin) is Di Bù Pi, and Er Jiao (er jiao). By studying weights and measures from the Ming and Qing dynasties, the relationship between the measurement and weight systems of the Ming and Qing dynasties and modern conversion systems was clarified. Volumes in the Ming and Qing dynasties were calculated in liters, dou, and hoku, with 1 hoku = 5 dou and 1 dou = 10 L, equivalent to 1 L of 1,035 mL today. Weights were calculated in liang and sen, with 1 liang = 10 sen, equivalent to 3.73 g today. Therefore, the above prescription for the two decoctions (daily dosage) was calculated as follows: Rehmannia Root 37.30 g, Xianthus Root 37.30 g, Peony Root 18.65 g, Wheat Dong Root 18.65 g, Digupi Root 11.19 g, and Er Jiao 11.19 g.

[0064] Example 2 Production of Ryochiyu standard samples and quality standards The original method for producing the two decoctions was "decocting with water," but there was no clear decoction method. This invention refers to the "Medical Institution Chinese Medicine Decoction Room Management Standards" and combines traditional decoction practices, considers and determines the soaking time, amount of water added, decoction strength, decoction time, concentration strength, and other important process parameters, determines the manufacturing process for the two decoctions reference sample, and establishes the quality standard for the two decoctions reference sample.

[0065] 1. Production of reference samples of Ryochiyu A 2L decoction pot was filled with five kinds of Chinese herbs: Rehmannia glutinosa 37.30g, Xanthan Ginseng 37.30g, Peony Root 18.65g, Wheat Flour 18.65g, and Dioscorea Root 11.19g. The pot was then covered and boiled twice. Seven times the amount of water as the five herbs was added to the first decoction, which was then steeped for 40 minutes. The pot was heated to a boil (power 2200W, 7 minutes), and then kept at a slight boil (power 400W) for 30 minutes (stirring once every 10 minutes, for at least 10 seconds each time). The pot was then filtered while still hot (through a 200-mesh sieve) to obtain the medicinal residue and the first decoction, ready for use. The resulting residue was added with five times the amount of water used for the five herbal medicines. The mixture was heated to a boil (2200W, approximately 5 minutes), then gently boiled (400W) for 20 minutes (stirring every 10 minutes for at least 10 seconds each time). The mixture was then filtered through a 200-mesh sieve while still hot to obtain the second decoction, which was then ready for use. The first and second decoctions were combined in a 2-L decoction pot and concentrated at atmospheric pressure (1200W) until the mixture reached approximately 500mL (approximately 20 minutes). 11.19g of Erjiao was added while still hot, and the mixture was stirred to dissolve, yielding a Liangchi Tang reference sample solution (20°C, density 1.05±0.01g / mL). The reference sample solution of both jiyuan was concentrated to approximately 250 mL under reduced pressure (T≦60°C) (concentration time: approximately 1 hour) to obtain a concentrated reference sample solution of both jiyuan (20°C, density: 1.07±0.01 g / mL). The concentrated reference sample solution of both jiyuan was freeze-dried for 48 hours (T≦-40°C, Vac≦20 Pa) to obtain a reference sample of both jiyuan (freeze-dried powder, moisture content: 4-7% by weight), which was sealed in a zippered storage bag and stored at room temperature in a desiccator.

[0066] 2 Quality standards for the Ryochiyu standard samples 2.1 Prescription (daily dose): Sake Dihuang 37.30g, Xuan Zheng 37.30g, Sake Baishai 18.65g, Mugidong 18.65g, Jiohide 11.19g and Agiao 11.19g. Description: This product is a tan to brown powder with a sweet, slightly bitter taste. 2.2 Characteristic spectrum Preparation of control substance solution: Appropriate amounts of paeoniflorin control substance and harpagoside control substance were taken, accurately measured, dissolved in methanol, and adjusted to volume to obtain control substance solutions, of which the concentration of paeoniflorin control substance was 200 μg / mL and the concentration of harpagoside control substance was 30 μg / mL. Preparation of test substance solution: The reference sample of both redigenous decoctions prepared in step (1) was collected and finely crushed. 0.5 g of the powder of both redigenous decoctions was accurately weighed and placed in a stoppered Erlenmeyer flask. 5 mL of water was accurately added to dissolve the powder. 20 mL of acetonitrile was accurately added and the flask was shaken while adding the water. After the addition, the flask was sealed and weighed. The flask was sonicated for 20 minutes (frequency 40 kHz, output 800 W), cooled (20°C), weighed again, and the weight loss was compensated for using 80% (v / v) aqueous acetonitrile solution. The filtrate was then accurately weighed and evaporated to dryness. 10% (v / v) aqueous methanol solution was added to the resulting residue and dissolved. The volume was adjusted to 10 mL to obtain the test substance solution.

[0067] Measurement: 10 μL of each of the control substance solution and the test substance solution was accurately taken and injected into a high performance liquid chromatograph for measurement, and the chromatogram was recorded for 60 minutes to obtain a control characteristic spectrum (see FIG. 1).

[0068] Chromatography conditions and system suitability test: Octadecylsilane-bonded silica was used as the packing material, and gradient elution was performed using methanol as mobile phase A and 0.05 v / v% aqueous phosphoric acid solution as mobile phase B according to the gradient elution procedure shown in Table 1. The detection wavelength was 254 nm, and the column temperature was 28 °C. The theoretical plate number was 20,000 to 30,000, calculated based on the paeoniflorin peak.

[0069] As shown in Figure 1, the characteristic spectrum of the test substance contains 10 characteristic peaks, two of which must match the retention times of the corresponding reference peaks. The peak corresponding to the paeoniflorin reference peak is the S peak. The relative retention times of each characteristic peak and the S peak are calculated, and the relative retention times must be within ±10% of the specified values: 0.22 (Peak 1), 0.26 (Peak 2), 0.31 (Peak 3), 0.33 (Peak 4), 1.00 (Peak 5), 1.47 (Peak 6), 1.54 (Peak 7), 1.82 (Peak 8), 1.91 (Peak 9), and 2.22 (Peak 10). Peaks 1–3 are unknown components, Peak 4 is gallic acid, Peak 5 is paeoniflorin, Peak 7 is verbascoside, Peak 8 is angoroside C, Peak 9 is cinnamic acid, and Peak 10 is harpagoside.

[0070] 2.3 Identification, moisture content and dry extractability Identification: (1) 4 g of this product powder was taken, 30 mL of methanol was added, and the mixture was sonicated for 30 minutes (400 W output), filtered, and the filtrate was concentrated to 5 mL and used as the test substance solution. 1.5 g of rhododendron bark was taken separately as a control, and a control solution was prepared in the same manner. 10 μL of each of the above two solutions was spotted on the same silica gel G thin-layer plate and developed using a 5:5:0.4 volumetric mixture of cyclohexane, ethyl acetate, and formic acid as the developing solvent. The plate was then removed, dried, and examined under a UV lamp (365 nm). The chromatogram of the test substance displayed fluorescent spots of the same color at the corresponding positions on the chromatogram of the control. Check: Moisture content is within the range of 3-8 wt%. The dry extraction rates ranged from 40.0 to 55.0%.

[0071] 2.4 Determination of harpagoside and paeoniflorin contents Chromatography conditions and system suitability test: A chromatography column packed with octadecylsilane-bonded silica was used, with methanol as mobile phase A and 0.05% v / v phosphoric acid aqueous solution as mobile phase B. The elution gradient procedure was as shown in Table 1. The detection wavelength was 254 nm, the column temperature was 28°C, and the theoretical plate number was 20,000 to 35,000, calculated based on the paeoniflorin peak.

[0072] Preparation of control substance solution: Paeoniflorin control substance and harpagoside control substance were accurately weighed, dissolved in methanol, and adjusted to volume to obtain a control substance solution, of which the harpagoside concentration was 20 μg / mL and the paeoniflorin concentration was 180 μg / mL. The test substance solution prepared in step 2.2 was used. Measurement: 10 μL of the control substance solution and the test substance solution were accurately taken and injected into a high performance liquid chromatograph for HPLC measurement to obtain the contents of harpagoside and paeoniflorin. Each dose (daily dose) contained 24-47 mg of Xianjin in terms of harpagoside and 198-370 mg of Jiangxiangxiangxiang in terms of paeoniflorin.

[0073] 2.5 Measurement of L-hydroxyproline and glycine content Chromatography conditions and system suitability test: A chromatography column using octadecylsilane-bonded silica as the packing material was used. Mobile phase A was acetonitrile-0.1 mol / L sodium acetate solution (adjusted to pH 6.5 with acetic acid) (acetonitrile:sodium acetate solution volume ratio = 7:93), and mobile phase B was 80% (v / v) acetonitrile aqueous solution. Gradient elution was performed according to the gradient elution procedure in Table 2. The flow rate was 0.8 mL / min, the detection wavelength was 254 nm, and the column temperature was 43 °C. The theoretical plate number based on the L-hydroxyproline peak was 56,000-10,000.

[0074] Preparation of control substance solution: Appropriate amounts of L-hydroxyproline control substance and glycine control substance were taken and accurately measured, and 0.1 mol / L hydrochloric acid solution was added to prepare a mixed solution containing 60 μg of L-hydroxyproline and 130 μg of glycine per mL.

[0075] Preparation of test substance solution: Approximately 1.5 g of this product powder was accurately weighed and placed in a 25 mL measuring flask. 20 mL of 0.1 mol / L hydrochloric acid solution was added. Sonication was performed for 30 minutes. Cooling was performed. 0.1 mol / L hydrochloric acid solution was added up to the line and the mixture was shaken uniformly. 2 mL was accurately weighed and placed in a 10 mL ampoule. 2 mL of hydrochloric acid was added. Hydrolysis was performed at 150°C for 1 hour. Cooling was performed. The mixture was then transferred to an evaporating dish and washed with 10 mL of water in batches. The washings were then evaporated to dryness in an evaporating dish. The residue was dissolved in 0.1 mol / L hydrochloric acid solution and transferred to a 25 mL measuring flask. 0.1 mol / L hydrochloric acid solution was added up to the line and the mixture was shaken uniformly.

[0076] Derivatization: Accurately measure 1 mL of each of the above control substance solution and test substance solution and place them in 25 mL stoppered test tubes. Add 0.5 mL of 0.1 mol / L phenylisothiocyanate (PITC) in acetonitrile and 0.5 mL of 1 mol / L triethylamine in acetonitrile, shake evenly, and leave at room temperature for 1 hour. Then add 3 mL of 50% (v / v) aqueous acetonitrile solution and shake evenly. Add 5 mL of n-hexane to each container, shake, and leave for 10 minutes. Filter the lower layer solution through a microporous membrane. The filtrates were the control substance derivatization solution and the test substance derivatization solution, respectively.

[0077] Measurement: 5 μL of the control substance derivatized solution and the test substance derivatized solution were accurately taken and injected into a high performance liquid chromatograph for measurement to obtain the contents of L-hydroxyproline and glycine.

[0078] Each dose of this product contains 0.64-1.18g of L-hydroxyproline and 1.26-2.33g of glycine.

[0079] 2.6 Functions and Indications: Nourishes yin, clears heat, nourishes blood, and regulates menstruation. Indicated for yin deficiency and blood-heat syndrome. Symptoms include early menstruation, redness, small amount, fever in the hands and feet, irritability, decreased sleep, red tongue with little tongue coating, and thready and rapid pulse.

[0080] Caution: Do not use in patients with severe fever or blood stasis or early menstruation, and should be used with caution in patients with weak spleen or stomach and soft stools. Storage: Keep tightly closed and store in a dry place.

[0081] Example 3 Research on the manufacturing process of Ryojito granules Ryoji tea drink pieces: Sake jiang 37.30g, Gensin 37.30g, Sakebaishai 18.65g, Mufufu 18.65g, Jibone skin 11.19g. 1. Research into the extraction process 1.1 Consideration of immersion time Seven portions of the two tea-drinking pieces were weighed and placed in seven 2-liter Erlenmeyer flasks. 750 mL of water was added to each flask, which were then tightly sealed and soaked at room temperature for 0, 30, 40, 50, 60, 70, 80, and 90 minutes. After soaking, the volume of the liquid remaining in each Erlenmeyer flask was measured and recorded as V. Water absorption curves were plotted, and the experiment was repeated twice. The water absorption results for the two tea-drinking pieces are shown in Table 3 and Figure 3.

[0082] Table 3 and Figure 3 show that the water absorption of the two geode tea pieces reaches an inflection point at 40 minutes, meaning that the water absorption is relatively high during the first 40 minutes, but decreases and stabilizes after 40 minutes. While a longer soaking time is beneficial for complete penetration of the geode tea pieces and for the release of ingredients during decoction, too long a soaking time can increase the risk of enzymatic hydrolysis of the medicinal liquid and mold and deterioration. After comprehensive consideration, a 40-minute soaking time was determined for the two geode tea pieces before decoction.

[0083] JPEG2025532254000003.jpg89170

[0084] 1.2 Consideration of number of extractions and extraction time Test method (two parallel tests were conducted for each test group): 1.5 times the weight of Liangjitang drinking pieces (Liangjitang Rehmannia Batch No. 20181210, Xuanzhen Batch No. 200501CP0184, Liangjiatang White Peony Batch No. 191101CP159, Mutiandu Batch No. 201909-001, Digupi Batch No. 20190312) were weighed and placed in a round-bottom flask. Water was added and the pieces were steeped at room temperature for 40 minutes, then heated to reflux and extracted three times (the water addition ratios and extraction times for the three extractions are shown in Table 4). The extracts were combined and sieved through a 200-mesh sieve for preparation. The extraction times and extraction times for the Liangjitang granules are shown in Table 5.

[0085] JPEG2025532254000004.jpg75170

[0086] JPEG2025532254000005.jpg105170

[0087] As shown in Table 5, (1) adding a third extraction to the extraction process of both granules increased the quality indicators, dry extraction rate, harpagoside, and paeoniflorin transfer rates, by 14.86%, 44.16%, and 29.98%, respectively. (2) Extending the extraction time increased the dry extraction rate, harpagoside, and paeoniflorin transfer rates by 12.31%, 19.72%, and 15.67%, respectively. (3) Increasing the number of extractions significantly improved the extraction rate, and extending the extraction time also improved the extraction rate. The dry extraction rate increased with the number of extractions, but the contents of both harpagoside and paeoniflorin decreased. With longer decoction times, harpagoside and paeoniflorin were partially destroyed by heat. (4) Preliminary analysis showed that adding a third extraction to both granules was superior, resulting in higher dry extraction rates, harpagoside, and paeoniflorin extraction rates.

[0088] 1.3 Consideration of extraction time and amount of water added Test method: The pieces of both tea leaves were weighed out in 80, 70, and 65 times the amount of each tea, and placed in a 100L integrated extractor / concentrator. Water was added, and the tea leaves were soaked at room temperature for 40 minutes, then heated and extracted at 99.5±0.5°C (extraction conditions are shown in Table 6). The extraction time and water addition test results are shown in Tables 7 and 8.

[0089] JPEG2025532254000006.jpg64170

[0090] JPEG2025532254000007.jpg62170

[0091] JPEG2025532254000008.jpg83170

[0092] As shown in Tables 7 and 8, (1) when the number of extractions was increased, the extraction time was extended (calculated from the results of the first three extractions), and the amount of water was increased, the main quality indicators, such as the dry extraction rate and the contents of harpagoside and paeoniflorin, all improved compared to the accompanying decoction (the reference sample from the same batch), but the differences between batches were not significant (RSD less than 5%). (2) When the extraction time was extended, the main quality indicators of the fourth extraction results, such as the dry extraction rate, harpagoside content, and paeoniflorin content, improved, but the extraction amount was only about 5%. (3) Verification tests were conducted with increased extraction times. Analysis of the results of the accompanying decoction (reference sample from the same batch) and the reference sample standard showed that the extraction rates (dry extraction rate 45.57%, harpagoside migration rate 70.10%, paeoniflorin migration rate 63.09%), which are the main quality indicators, reached the extraction targets of the two granules, were within the standard range of the reference sample, and were higher than those of the accompanying decoction (reference sample from the same batch).

[0093] 1.4 Dissolution test of paeoniflorin and harpagoside The extraction process of traditional Chinese medicine involves dissolving each component in a solvent. To further verify the rationality of the extraction process of Liangjitang Granules, paeoniflorin and harpagoside were used as representative components, and the tendency of the content changing with the extraction time was tracked and considered. An extraction time-integral area curve was drawn to determine the optimal extraction time.

[0094] Test method: Two portions of the two decoction pieces, each 1.5 times the weight of the original, were weighed and placed in a round-bottom flask. Seven times the volume of water was added and heated using an electric heating jacket. Once boiling, the time was measured and the mixture was maintained at a gentle boil for 120 minutes. Five milliliters of the solution were sampled every 10 minutes using a pipette and filtered through a microporous membrane. The paeoniflorin and harpagoside contents in the two decoction reference samples were then measured. Based on the results, the dissolution curves (extraction time-integrated area) of the two components, which varied with extraction time, were plotted. The changes in the paeoniflorin and harpagoside contents in the decoction at different extraction times were compared to determine the trends of change. The dissolution curves for harpagoside and paeoniflorin are shown in Figure 4 and Figure 5, respectively.

[0095] As shown in Figures 4 and 5, when extracted from Liangji Tang for 30 minutes, harpagoside and paeoniflorin accounted for 90% and 78% of the total extracted amount, respectively. The extraction process (3.3.4.4.1) preferably selected based on the extraction time considerations for Liangji Granules (batch number LDTXS2102003) had a higher extraction rate than the optimal extraction time (single extraction, adding 7 times the amount of water, 70 minutes), further verifying that the extraction parameters were reasonable.

[0096] 2. Concentration process research Vacuum evaporation is a commonly used concentration method for modern traditional Chinese medicine formulations, with the advantages of low temperature and high efficiency. Liangji granules were concentrated by vacuum evaporation, and a reasonable vacuum concentration temperature range for Liangji Tang formulations was determined through rotary evaporation experiments.

[0097] Test method: Weigh out 2.6 kg of Rehmannia glutinosa liquor, 2.6 kg of Xanthan Ginseng liquor, 1.3 kg of Peony liquor, 1.3 kg of Wheat Flour, and 0.78 g of Dioscorea Root Bark into a 100L extraction tank, add water, and boil three times. The first time, add 7 times the amount of water and steep for 40 minutes, then heat and extract for 30 minutes. The second time, add 5 times the amount of water and steep for 20 minutes. The third time, add 5 times the amount of water and steep for 20 minutes. The extracts are combined and ready for use.

[0098] Approximately 4 kg of the combined extract was taken and divided into four portions. Each portion was weighed and placed in a rotary evaporator. The liquid was concentrated to a relatively high concentration (approximately 50 g) at temperatures of 60°C, 70°C, 80°C, 90°C, and 95°C. The concentration time and degree of vacuum were recorded, and the contents of harpagoside and paeoniflorin in the concentrated liquid were determined. The results showed that the optimal temperature for vacuum concentration of both granules was between 70°C and 90°C. The test results are shown in Table 9.

[0099] JPEG2025532254000009.jpg59170

[0100] As shown in Table 9, when the extract of Ryoji Tang granules was concentrated under reduced pressure at temperatures between 60 and 90°C, there was almost no difference in the quality indicators of harpagoside and paeoniflorin (RSD < 3.0%). However, as the concentration temperature and concentration efficiency increased, the optimal temperature for reduced-pressure concentration of Ryoji granules was in the range of 70 to 90°C. When concentrated under reduced pressure at 95°C, the temperature was too high, making it prone to severe boiling and backflow.

[0101] 3 Drying process research Vacuum belt drying has the advantages of low drying temperature, short drying time, minimal loss, and high efficiency. It is particularly suitable for materials that are viscous, prone to aggregation, and heat-sensitive. The formulation of the two-layer granules contains Rehmannia glutinosa, Barley Winter, and Xanthan Ginseng, which contain relatively high amounts of polysaccharides and have a high viscosity. Therefore, vacuum belt drying was selected as the drying method for the two-layer granules.

[0102] There are many factors that affect vacuum belt drying, mainly the density of the chemical solution, the feed rate, the drying time (i.e., the speed of the crawler belt), and the temperature of the heating zone. Therefore, these important parameters were considered to determine the drying process of both granules.

[0103] 3.1 Consideration of chemical solution density Mixed concentrates of the two granules with different concentrations (density 1.26-1.35 g / mL) were taken and dried and powdered in a vacuum belt dryer. The test process was observed, the powder output amount was measured, and the powder output rate was calculated.

[0104] The results showed that when the density of the dry liquid for the double-ground granules was in the range of 1.26 to 1.35 g / mL, smooth dispensing, uniform spraying, minimal adhesion, and good dry powder quality were achieved. When the density of the dry liquid was 1.26 g / mL, the coating of the material liquid was thin, resulting in a slight decrease in drying efficiency. When the density of the liquid was 1.36 g / mL, the viscosity of the material liquid was high, allowing for dispensing, but swelling during the drying process resulted in losses. Therefore, the optimal density for the dry liquid for the double-ground granules was 1.26 to 1.35 g / mL.

[0105] 3.2 Drying temperature considerations Due to the characteristics of the vacuum belt dryer, the drying temperatures for the herbal extracts were 95±5°C in the first stage, 96±5°C in the second stage, and 96±5°C in the third stage. A drying test was conducted on the concentrate of Liangji Tang (batch number). The results showed that under these drying temperature conditions, the main quality indicators of Liangji Granules, paeoniflorin and harpagoside, were relatively stable. Therefore, the belt drying temperature conditions for Liangji Granules were set at 95±5°C in the first stage, 96±5°C in the second stage, and 96±5°C in the third stage.

[0106] Example 4 Experimental study on the stability of paeoniflorin 1. Purpose of the experiment The problem of paeoniflorin content loss was investigated based on the Liangjiang formulation process. Analysis revealed that the loss of paeoniflorin was related to the decomposition of paeoniflorin due to prolonged vacuum concentration, and the influence of the addition of ermine gelatin and dextrin on the dissolution of paeoniflorin. In this experiment, single-factor experiments were used to examine the effects of heating time and paeoniflorin concentration on paeoniflorin content. At the same time, comparative experiments were used to examine the effects of the addition of ermine gelatin and dextrin on the dissolution of paeoniflorin.

[0107] II. Experimental Method 2.1 Experimental study on the thermal stability of paeoniflorin 2.1.1 Preparation of sample solution A certain amount of paeoniflorin control substance was taken, accurately weighed, and dissolved in methanol to obtain a paeoniflorin solution with a concentration of 1.8905 mg / mL, which was used as the mother solution. An appropriate amount of the mother solution was weighed and purified water was added to prepare paeoniflorin solutions with different concentrations (serial numbers 1 to 5) as shown in Table 10. Paeoniflorin solutions 1 to 5 were placed in a water bath and heated to 85±5°C. After heating for 0.5 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 3.5 hours, and 4.0 hours, sampling was performed to obtain sample solutions.

[0108] JPEG2025532254000010.jpg43170

[0109] 2.1.2 Preparation of test substance solutions An appropriate amount of each sample solution prepared in 2.1.1 was taken and purified water was added to prepare test substance solutions No. 1 to No. 5, which were then sampled and detected. The method for preparing test substance solutions No. 1 to No. 5 is as follows. Series 1: Paeoniflorin solution with a concentration of 0.05 mg / mL was collected at different heating times (0.5 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 3.5 hours, and 4.0 hours) and filtered through a 0.45 μm filter membrane. Series 2: 0.3 mg / mL paeoniflorin solution was accurately taken at 0.3 mL each at different heating times (0.5 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 3.5 hours, and 4.0 hours), and 0.7 mL of purified water was added. The mixture was shaken evenly and filtered through a 0.45 μm filter membrane. Series 3: 0.2 mL of 0.5 mg / mL paeoniflorin solution was accurately taken at different heating times (0.5 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 3.5 hours, and 4.0 hours), and 0.8 mL of purified water was accurately added. The mixture was shaken evenly and filtered through a 0.45 μm filter membrane. Series 4: 0.1 mL of 0.8 mg / mL paeoniflorin solution was accurately taken, 0.9 mL of purified water was accurately added, the solution was shaken evenly, and filtered through a 0.45 μm filter membrane. Series 5: 0.1 mL of 1.0 mg / mL paeoniflorin solution was accurately taken at different heating times (0.5 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 3.5 hours, and 4.0 hours), placed in a 2 mL measuring flask, and purified water was added up to the mark. The flask was shaken evenly and filtered through a 0.45 μm filter membrane.

[0110] 2.1.3 Detection Method Chromatography conditions and system suitability test: A chromatography column using octadecylsilane-bonded silica as the packing material was used, and acetonitrile-0.1% phosphoric acid solution (volume ratio 14:86) was used as the mobile phase. The detection wavelength was 230 nm, the column temperature was 27°C, the mobile phase flow rate was 1.0 mL / min, and the theoretical plate number calculated based on the paeoniflorin peak was 28,000. Measurement: 10 μL of each of the test substance solutions from Series 1 to Series 5 prepared in 2.1.2 was accurately sampled and injected into a high-performance liquid chromatograph for measurement, resulting in the liquid phase spectra of paeoniflorin in the test substance solutions from Series 1 to Series 5. The liquid phase spectra of paeoniflorin in the test substance solutions from Series 1 to Series 5 are shown in Figure 6, Figure 7, Figure 8, Figure 9, Figure 10, Figure 11, Figure 12, Figure 13, Figure 14, Figure 15, Figure 16, Figure 17, Figure 18, Figure 19, Figure 20, Figure 21, Figure 22, Figure 23, Figure 24, Figure 25, Figure 26, Figure 27, Figure 28, Figure 29, Figure 30, Figure 31, Figure 32, Figure 33, Figure 34, Figure 35, Figure 36, Figure 37, Figure 38, Figure 39, Figure 40, Figure 41, Figure 42, Figure 43, Figure 44, Figure 45, Figure 46, Figure 47, Figure 48, Figure 49, Figure 50, Figure 51, Figure 52, Figure 53, Figure 54, Figure 55, Figure 56, Figure 57, Figure 58, Figure 59, Figure 60, Figure 61, Figure 62, Figure 63, Figure 64, Figure 65, Figure 66, Figure 67, Figure 68, Figure 69, Figure 70, Figure 71, Figure 72, Figure 73, Figure 74, Figure 75, Figure 76, Figure 77, Figure 78, Figure 79, Figure 80, Figure 81, Figure 82, Figure 83, Figure 84, Figure 85, Figure 86, Figure 87, Figure 88,

[0111] 2.1.4 Data analysis The horizontal axis represents heating time and the vertical axis represents paeoniflorin area %. Curves of different concentrations of paeoniflorin solutions that change with heating time were created to analyze the effects of concentration and heating time on the stability of paeoniflorin. 2.2 Consideration of the effect of dextrin and gelatin on the dissolution of paeoniflorin 2.2.1 Experimental method Preparation of sample solution: 0.53 mL of paeoniflorin mother solution (1.890 mg / mL) was accurately measured and placed in a 50 mL volumetric flask. Water was added up to the measuring line and the flask was shaken evenly to obtain a paeoniflorin solution with a concentration of 0.02 mg / mL. Appropriate amounts were placed in stoppered test tubes, and dextrin and gelatin were added. Each tube was treated using different methods (see Table 11). The mixture was mixed evenly, the pH was measured, the tube was sealed, and the mixture was heated at 85°C for 48 hours to obtain the sample solution. Samples were taken at 4, 22, 32, and 48 hours after heating to determine the paeoniflorin content.

[0112] JPEG2025532254000011.jpg80170

[0113] Preparation of test substance solution: 2 mL of sample solution was accurately taken and placed in a stoppered Erlenmeyer flask, 8 mL of acetonitrile was accurately added, the flask was shaken evenly, weighed, sealed, and ultrasonically treated (800 W, 40 kHz) for 20 minutes. After cooling, the flask was weighed again, and the lost weight was compensated for using an 80% (v / v) aqueous acetonitrile solution. The flask was shaken evenly and filtered through a 0.45 μm filter membrane, and the resulting filtrate was used as the test substance solution.

[0114] 2.2.3 Detection method: The detection was carried out according to 2.1.3. 2.2.4 Data analysis The concentration of paeoniflorin in the samples was calculated using equation (1), and the concentrations of paeoniflorin in different samples were compared to calculate the RSD%. Paeoniflorin concentration in sample C = measured amount (μg) / test substance loading amount (μL) × constant volume of test substance solution (mL) / volume of test substance solution (mL) Formula (1).

[0115] 3. Experimental results 3.1 Consideration test on the thermal stability of paeoniflorin The peak area of ​​each sample was obtained by detecting samples of a series of paeoniflorin solutions with different concentrations at different heating times. The initial (0 h) peak area of ​​each series was set to 100, and the peak integral relative value of each sample was calculated by dividing the initial peak area of ​​the other samples by the initial peak area of ​​that series. The detection results are shown in Table 12. A time-relative peak area change graph was drawn, with the heating time on the horizontal axis and the relative value of the peak integral area of ​​paeoniflorin in each concentration of paeoniflorin solution on the vertical axis. The results are shown in Figure 11, which shows the test results of paeoniflorin solutions with different concentrations changing with heating time. To further investigate the relationship between the stability of paeoniflorin solution and its concentration and heating time, we analyzed the chromatograms of paeoniflorin solution with heating time and those of paeoniflorin solutions of different concentrations at the same heating time. The results are shown in Figures 12 and 13. Figure 12 shows the chromatogram of a 0.05 mg / mL paeoniflorin solution after heating for 4 hours. The heating times, from bottom to top, are 0 hours, 0.5 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, and 4 hours. Figure 13 shows the chromatogram of paeoniflorin solutions of different concentrations after heating for 4 hours. The concentrations, from bottom to top, are 0.05 mg / mL, 0.3 mg / mL, 0.5 mg / mL, 0.8 mg / mL, and 1.0 mg / mL.

[0116] JPEG2025532254000012.jpg108170

[0117] Table 12 and Figures 11-13 show that the stability of paeoniflorin during the heating process is related to the concentration of the paeoniflorin solution. After 4 hours of heating, the paeoniflorin peak areas for the 0.05 mg / mL, 0.3 mg / mL, 0.5 mg / mL, 0.8 mg / mL, and 1.0 mg / mL paeoniflorin solutions decreased to 73.43%, 85.66%, 95.39%, 96.69%, and 96.89% of their initial values, respectively. Two impurity peaks were clearly observed in the low-concentration (<0.5 mg / mL) chromatograms. The areas of the impurity peaks increased with increasing heating time, and the areas of the impurity peaks decreased with increasing paeoniflorin solution concentration. This indicates that the degradation of paeoniflorin during the heating process is related to the heating time and paeoniflorin concentration. The longer the heating time, the lower the concentration and the more severe the degradation.

[0118] 3.2 Consideration of the effect of dextrin and gelatin on the dissolution of paeoniflorin The concentration of paeoniflorin in the samples was calculated, and the concentrations of paeoniflorin in different samples were considered in parallel, and the RSD% was calculated, and the results were shown in Table 13.

[0119] JPEG2025532254000013.jpg98170

[0120] As shown in Table 13, there was no significant difference between the paeoniflorin concentration in the sample to which PEG-1444 and dextrin had been added and the paeoniflorin concentration in the undiluted sample (RSD%<5.0).

[0121] 4. Experimental conclusions 1. The decomposition of paeoniflorin during the heating process of paeoniflorin solution is related to heating time and paeoniflorin concentration. The longer the heating time, the lower the paeoniflorin concentration and the more severe the paeoniflorin degradation. Further analysis revealed that paeoniflorin concentration is related to pH. Paeoniflorin contains a large number of hydroxyl groups, and the higher the concentration, the lower the pH. Research has shown that paeoniflorin is more stable in an acidic environment with a pH of 3 to 5. Above a pH of 6, paeoniflorin stability significantly decreases. The pH of the extract of the two herbs is approximately 6, and paeoniflorin becomes unstable during the concentration process. Therefore, paeoniflorin decomposes when concentrated under reduced pressure. 2. The addition of PEG-404 and dextrin did not significantly affect the release of paeoniflorin (RSD%<5), indicating that the embedding of PEG-404 and dextrin did not affect the release of paeoniflorin. 3. The decomposition of paeoniflorin is mainly related to the pH of the medicinal solution, and the pH of the medicinal solution needs to be controlled during the extraction and concentration process of the two decoctions. Further research is needed on the specific methods for controlling pH.

[0122] Example 5 Manufacturing and quality standards for Ryojito granules 1. Manufacturing of Ryojito granules Soaking: 208.0 kg of Rehmannia glutinosa root, 208.0 kg of Xanthan Ginseng root, 104.0 kg of White Peony root, 104.0 kg of Wheat Flour, and 62.4 kg of Dioscorea Root were weighed and placed in an extraction tank. Water 7 times the amount of the pieces was added, and the pieces were soaked for 40 minutes. Extraction: For the first extraction, the tea was heated to 100°C and extracted for 30 minutes to release the medicinal liquid. For the second extraction, five times the amount of water was added, the tea was heated to 100°C and extracted for 20 minutes to release the medicinal liquid. For the third extraction, five times the amount of water was added, the tea was heated to 100°C and extracted for 20 minutes to release the medicinal liquid. Centrifugation: Each chemical solution was centrifuged using a centrifuge (separation speed: 60 L / min, slug discharge: 10 min / time). Microfiltration: The centrifuged solution from each run was filtered through a plate and frame filter (membrane pore size: 5-15 μm). Dissolving the gelatin glue: Weigh out 62.4 kg of gelatin glue and place it in a sandwich pot. Add water four times the mass of the gelatin glue and heat until it boils. Stir constantly to dissolve the gelatin glue until it becomes a homogeneous liquid. Remove the floating bubbles and impurities on top and pass it through a 100-mesh drying net to obtain the gelatin glue liquid, which is ready for use. Dissolving dextrin: 40 kg of dextrin was weighed and poured into a stainless steel barrel, and water four times the mass of the dextrin was added and stirred until completely dissolved to obtain an aqueous dextrin solution, which was then ready for use. Batch concentration of extract: Turn on the vacuum and suck the filtered liquid into the vacuum concentrator of the concentrator in each batch. Open the steam valve to start heating, control the temperature to 85±5℃, vacuum to 0.045±0.015MPa, and steam pressure to 0.04±0.01MPa. Monitor the boiling condition and temperature from time to time, adjust the vacuum and steam pressure, and concentrate the liquid density to 1.10g / mL or more but less than 1.20g / mL. Then, put them into liquid storage tanks / transport container barrels and weigh them for use. Mixing and concentration: Turn on the vacuum and suck the triple-extracted concentrate, glue solution, and dextrin aqueous solution into the single-effect concentrator. Continue concentrating in batches until a thick paste with a density of 1.25-1.36 g / mL (20°C) is obtained. Then, place the mixture in a transport container barrel, weigh it, and prepare it for use. Drying: The vacuum belt dryer was turned on to dry and powderize the powder. The parameters were as follows: the feed temperature was 80°C, the feed rate was 18 L / h, the crawler belt speed was 16±5 cm / min, and the swing arm angle was 70°. The cloth motor speed was 40 r / min. The first, second, third, and fourth heating temperatures were 95±5°C, 96±5°C, 96±5°C, and 30±5°C (cooling), respectively. The cutter was turned on for 10 seconds, and the vacuum was 0.097-0.1 MPa. The powder was collected in a transport container barrel, sealed, and ready for use. Granulation: The dry powder was taken, weighed and transferred to the granulator. The roller pressure of the granulator was adjusted to 16 MPa, and the screw feeder rotation speed was adjusted to a vertical rotation speed of 24 r / min, a horizontal rotation speed of 120 r / min, a roller rotation speed of 6 r / min and a granulation speed of 130 r / min to operate the granulation. During operation, the state of the equipment and the properties of the granules were observed, the granulation parameters were adjusted appropriately, powder was added in a timely manner to collect the granules, and the adjustment status of the granulation parameters was recorded. The vibrating screen was turned on, and the granulated granules were placed in the hopper. The hopper valve was adjusted so that the feed rate was fast enough to completely separate the fine powder from the granules. The granules were placed in a transfer container barrel, and the fine powder was re-granulated in preparation for use, producing a total of 400 kg of Liangjitang granules. Packaging: Packaged using medicinal composite film at 12g per bag, a total of 330,000 bags have been produced.

[0123] 2. Quality standards for Ryochito granules 2.1 Prescription: Sake Dihuang 520g, Genshen 520g, Sake Baishai 260g, Mugidong 260g, Jibone Peel 156g and Agiao 156g. Description: The product appears as light to dark gray granules. It has a sweet and slightly bitter taste. 2.2 Characteristic spectrum Preparation of control substance solution: Appropriate amounts of paeoniflorin control substance and harpagoside control substance were taken, accurately measured, dissolved in methanol, and adjusted to volume to obtain control substance solutions, of which the concentration of paeoniflorin control substance was 200 μg / mL and the concentration of harpagoside control substance was 30 μg / mL. Preparation of test substance solution: Finely crush the Liangchi Tang granules prepared in step (1) and weigh approximately 1 g. Place in a stoppered Erlenmeyer flask, add 1 g of diatomaceous earth, mix thoroughly, add 25 mL of 80% v / v acetonitrile, and weigh. Sonicate for 20 minutes (power output 800 W, frequency 40 kHz). Cool and reweigh. Add 80% v / v acetonitrile to replace the lost weight. Shake thoroughly and filter. Accurately measure 15 mL of the filtrate, evaporate to dryness, and dissolve the residue in 10% methanol. Transfer to a 10 mL volumetric flask, add 10% methanol (v / v) up to the line, shake thoroughly, and filter. The filtrate was used to obtain the test substance solution. Measurement: 10 μL of each of the control substance solution and the test substance solution was accurately taken and injected into a high performance liquid chromatograph for measurement, and the chromatogram was recorded for 60 minutes to obtain a control characteristic spectrum (see FIG. 2). Chromatography conditions and system suitability test: Octadecylsilane-bonded silica was used as the packing material, and gradient elution was performed using methanol as mobile phase A and 0.05 v / v% aqueous phosphoric acid solution as mobile phase B according to the gradient elution procedure shown in Table 1. The detection wavelength was 254 nm, and the column temperature was 28°C. The theoretical plate number was 30,100, calculated based on the paeoniflorin peak.

[0124] As shown in Figure 2, the characteristic spectrum of the test substance showed 10 characteristic peaks, two of which must match the retention times of the corresponding reference peaks. The peak corresponding to the paeoniflorin reference peak was the S peak. The relative retention times of each characteristic peak and the S peak were calculated, and the relative retention times must be within ±10% of the specified values. The specified values ​​were 0.22 (Peak 1), 0.26 (Peak 2), 0.31 (Peak 3), 0.33 (Peak 4), 1.00 (Peak 5), 1.47 (Peak 6), 1.54 (Peak 7), 1.82 (Peak 8), 1.91 (Peak 9), and 2.22 (Peak 10). Peaks 1–3 were unknown peaks, Peak 4 was gallic acid, Peak 5 was paeoniflorin, Peak 7 was verbascoside, Peak 8 was angoroside C, Peak 9 was cinnamic acid, and Peak 10 was harpagoside.

[0125] 2.3 Moisture content and dry extraction rate Identification: (1) 4 g of this product powder was added to 30 mL of methanol, sonicated for 30 minutes (power output 400-600 W), filtered, and the filtrate was concentrated to 5 mL and used as the test substance solution. 1.5 g of rhododendron bark was used as a control, and a control solution was prepared according to the method for preparing the test substance solution. 10 μL of each of the above two solutions was spotted on the same silica gel G thin-layer plate and developed using a 5:5:0.4 volumetric mixture of cyclohexane, ethyl acetate, and formic acid as the developing solvent. The plate was then removed, dried, and examined under a UV lamp (365 nm). The chromatogram of the test substance displayed fluorescent spots of the same color at the corresponding positions on the chromatogram of the control. Check: The dry extraction rate was in the range of 40.0-55.0%.

[0126] 2.4 Determination of harpagoside and paeoniflorin contents Chromatography conditions and system suitability test: A chromatography column using octadecylsilane-bonded silica as the packing material was used, with methanol as mobile phase A and 0.05 v / v% aqueous phosphoric acid as mobile phase B. The elution gradient procedure was as shown in Table 1. The detection wavelength was 254 nm, the column temperature was 28°C, and the theoretical plate number was 2910, calculated based on the paeoniflorin peak. Preparation of control substance solution: Paeoniflorin control substance and harpagoside control substance were accurately weighed, dissolved in methanol, and adjusted to volume to obtain a control substance solution, of which the harpagoside concentration was 30 μg / mL and the paeoniflorin concentration was 200 μg / mL. The test substance solution prepared in step 2.2 was used. Measurement: 10 μL of the control substance solution and the test substance solution were accurately taken and injected into a high performance liquid chromatograph for HPLC measurement to obtain the contents of harpagoside and paeoniflorin. One bag (12g) of Ryojito granules contained 4.0-8.0mg of Xianjin in terms of harpagoside and 33.0-62.0mg of Jiangxiangxiang in terms of paeoniflorin.

[0127] 2.5 Measurement of gelatin gelatin (L-hydroxyproline and glycine) content Chromatography conditions and system suitability test: A chromatography column using octadecylsilane-bonded silica as the packing material was used. Mobile phase A was acetonitrile-0.1 mol / L sodium acetate solution (adjusted to pH 6.5 with acetic acid) (acetonitrile:sodium acetate solution volume ratio = 7:93), and mobile phase B was 80% (v / v) acetonitrile aqueous solution. Gradient elution was performed according to the gradient elution procedure in Table 2. The flow rate was 0.8 mL / min, the detection wavelength was 254 nm, and the column temperature was 43 °C. The theoretical plate number based on the L-hydroxyproline peak was 6,100. Preparation of control substance solution: Appropriate amounts of L-hydroxyproline control substance and glycine control substance were taken and accurately measured, and 0.1 mol / L hydrochloric acid solution was added to prepare a mixed solution containing 60 μg of L-hydroxyproline and 130 μg of glycine per mL. Preparation of test substance solution: Approximately 1.5 g of this product powder was accurately weighed and placed in a 25 mL measuring flask. 20 mL of 0.1 mol / L hydrochloric acid solution was added. Sonication was performed for 30 minutes. Cooling was performed. 0.1 mol / L hydrochloric acid solution was added up to the line and the mixture was shaken uniformly. 2 mL was accurately weighed and placed in a 10 mL ampoule. 2 mL of hydrochloric acid was added and the mixture was hydrolyzed at 150°C for 1 hour. Cooling was performed. The mixture was transferred to an evaporating dish and washed with 10 mL of water in batches. The washings were then evaporated to dryness in an evaporating dish. The residue was dissolved in 0.1 mol / L hydrochloric acid solution and transferred to a 25 mL measuring flask. 0.1 mol / L hydrochloric acid solution was added up to the line and the mixture was shaken uniformly. Derivatization: Accurately measure 1 mL of each of the above control substance solution and test substance solution and place them in 25 mL stoppered test tubes. Add 0.5 mL of 0.1 mol / L phenylisothiocyanate (PITC) in acetonitrile and 0.5 mL of 1 mol / L triethylamine in acetonitrile, shake evenly, and leave at room temperature for 1 hour. Then add 3 mL of 50% (v / v) aqueous acetonitrile solution and shake evenly. Add 5 mL of n-hexane to each solution, shake, and leave for 10 minutes. Filter the lower layer solution through a microporous membrane. The filtrates were the control substance derivatization solution and the test substance derivatization solution, respectively. Measurement: 5 μL of the control substance derivatized solution and the test substance derivatized solution were accurately taken and injected into a high performance liquid chromatograph for measurement to obtain the contents of L-hydroxyproline and glycine. The content of Erjiao in one bag (12 g) of Ryochito granules was 0.10-0.20 g in terms of L-hydroxyproline and 0.21-0.39 g in terms of glycine.

[0128] 2.6 Functions and Indications: Nourishes yin, clears heat, nourishes blood, and regulates menstruation. Indicated for yin deficiency and blood-heat syndrome. Symptoms include early menstruation, redness, small amount, fever in the hands and feet, irritability, decreased sleep, red tongue with little tongue coating, and thready and rapid pulse. Dosage and Administration: Take two packets three times a day with hot water. Specifications: 12g per bag. Storage: Keep tightly closed and store in a dry place.

[0129] Example 6 Three consecutive batches of commercial-scale production trials of Ryojito granules 1 Testing process (1) Soaking, extraction 208.0 kg of liquor Rehmannia glutinosa, 208.0 kg of Xanthan Ginseng, 104.0 kg of liquor White Peony, 104.0 kg of Wheat Flour, and 62.4 kg of Dioscorea officinalis were weighed, divided into two equal parts, and placed in two extraction tanks. Each tank was filled with 7 times the amount of water the pieces had, and the pieces were steeped for 40 minutes. In the first batch, 5 times the amount of water the pieces had was added, and the pieces were heated to 100°C and steeped for 20 minutes to release the medicinal liquid. In the second batch, 5 times the amount of water the pieces had been added, and the pieces were heated to 100°C and steeped for 20 minutes to release the medicinal liquid. In the third batch, 4 to 5 times the amount of water the pieces had been added, and the pieces were heated to 100°C and steeped for 20 minutes to release the medicinal liquid. (2) Centrifugation and microfiltration Each chemical solution was centrifuged (separation speed 60 L / min, slug discharge 10 min / cycle), and the number of slug discharges was recorded. Each centrifuged solution was filtered through a plate-frame filter (membrane pore size 5-15 μm), and the number of plate changes was recorded (the fine filter plate was replaced when the pressure exceeded 0.2 MPa). (3) Concentration The vacuum was turned on, and the filtered liquid was sucked into the vacuum concentrator of the concentrator in each batch. The steam valve was opened to start heating, and the temperature was controlled at 85±5°C, the vacuum at 0.055±0.015MPa, and the steam pressure at ≦0.09MPa. The boiling condition and temperature were monitored at regular intervals, and the vacuum and steam pressure were adjusted to concentrate the liquid until its density reached ≧1.10g / mL (20°C), which was then placed in turn into a liquid storage tank / transport container barrel and weighed for use. (4) Dissolving glue and dextrin Weigh out 62.4 kg of Egashira and put it into a sandwich pot. Add water in an amount six times the mass of Egashira. Heat to a boil and stir constantly to dissolve into a homogeneous Egashira solution. Remove the floating bubbles and impurities on top and pass it through a 120 mesh drying net to obtain the Egashira solution, which is ready for use. 40 kg of dextrin (10% of the dry extract) was weighed out, placed in a stainless steel barrel, and water four times the mass of the dextrin was added and stirred until completely dissolved to obtain a dextrin solution, which was then ready for use. (5) Mixing, concentration The three concentrates, the glue solution, and the dextrin solution were sucked into the single-effect concentrator and continued to be concentrated using the concentration conditions in step (3) until a thick paste with a density of 1.25-1.36 g / mL (20°C) was obtained, which was then placed in a transport container barrel, weighed, and prepared for use. (6) Drying The mixed concentrate was placed in a liquid storage tank, heated to 90±5°C, sterilized for 60 minutes, and cooled to 80±5°C. A vacuum belt dryer was turned on to dry and powderize the mixture. The parameters were as follows: feed temperature: 83±2°C, feed rate: 17.5±2.5 L / h, crawler belt speed: 16±5 cm / min, swing arm angle: 80°. The fabric motor speed was 40 r / min. The first, second, third, and fourth heating temperatures were 95±5°C, 96±5°C, 96±5°C, and 30±5°C (cooling), respectively. The cutter was used for 20 seconds per cycle, and the vacuum pressure was 0.097-0.1 MPa. The powder was collected in a transport container barrel, sealed, and ready for use. (7) Granulation The dry powder was taken, weighed, and transferred to a granulator. The roller pressure was adjusted to 17 MPa, and the screw feeder rotation speeds were adjusted to 20 r / min vertically, 100 r / min horizontally, 5.5 r / min rollers, and 130 r / min granulation speed. During operation, the equipment status and granule characteristics were observed, and the granulation parameters were adjusted appropriately. Powder was added and collected as needed. The vibrating screen was turned on, and the granules were placed in the hopper. The hopper valve was adjusted to ensure that the feed rate was sufficient to completely separate the fine powder from the acceptable granules. The granules were placed in the transfer container barrel for use, and the fine powder was re-granulated. The granulation parameter adjustments were recorded. Granules were sampled to determine their moisture content, paeoniflorin, and harpagoside content. (8) Individual packaging Each 12g bag was packaged using a composite film, and the filling amount for each 12g bag was controlled between 12.00g and 12.50g (the packaging yield was calculated, and samples were taken to detect particle size, moisture content, paeoniflorin content, harpagoside content, characteristic spectrum, etc.).

[0130] 2. Test Results In three consecutive batches of commercial-scale production testing of Liangjitang granules, the process route was reasonable, the main equipment such as extraction, centrifugation, microfiltration, concentration, drying and granulation were properly combined, each major process parameter was within the control range, the entire testing process was relatively smooth, and the production volume and quality indicators of the three batches of Liangjitang granules produced all met the quality control standards of Liangjitang preparations, with small differences (RSD<5% for dry extraction rate, paeoniflorin content and harpagoside content of the three batches of finished product). The results are shown in Table 14.

[0131] JPEG2025532254000014.jpg179170

[0132] Example 7 1. Pilot test of 3 batches (mixed concentration test) (batch numbers 2105012, 2105013, 2106014) 1.1 Three-batch pilot test method 12.309 kg of Rehmannia glutinosa root, 12.309 kg of Xanthan Ginseng root, 6.138 kg of White Peony root, 6.138 kg of Wheat Flour, and 3.696 kg of Dioscorea Root were weighed out, and water seven times the volume of the pieces was added. The pieces were then steeped for 40 minutes. The first steeping was performed by heating at 100°C for 30 minutes to extract the medicinal liquid. The second steeping was performed by adding water five times the volume of the pieces, heating at 100°C for 20 minutes to extract the medicinal liquid. The third steeping was performed by adding water five times the volume of the pieces, heating at 100°C for 20 minutes to extract the medicinal liquid. The three steepings were measured, and the density, dry extractability, harpagoside, and paeoniflorin were detected. Each drug solution was filtered through a plate and frame filter (membrane pore size: 5-15 μm), the density of the drug solution was detected (the fine filter plate was replaced when the pressure exceeded 0.02 MPa), and the number of times the plate was replaced was recorded. 3.696 kg of gelatinous gelatin was weighed out, powdered, and six times the amount of water as the gelatinous gelatin was added. The mixture was heated to a boil and stirred constantly to dissolve the gelatinous gelatin into a homogeneous solution. The concentration was continued until the densities reached 1.16 g / mL, 1.18 g / mL, and 1.17 g / mL. 2.4 kg of dextrin was weighed out and placed in a stainless steel barrel, and water four times the mass of the dextrin was added and stirred until completely dissolved, to obtain a dextrin solution. The vacuum was turned on and the filtered solution was sucked into the vacuum concentrator. The steam valve was opened to begin heating, controlling the temperature at 85±5°C, the vacuum at 0.045±0.015 MPa, and the steam pressure at 0.025±0.005 MPa. The boiling and temperature were monitored as needed, and the vacuum and steam pressure were adjusted to concentrate the solution to a density of approximately 1.20 g / mL. The gelatin and dextrin solutions were then sucked in, and the concentration continued until a thick paste with a density of 1.30-1.35 g / mL (20°C) was obtained. The concentrate was then placed in a transport container barrel for use. The concentrate was weighed and analyzed for density, dry extractability, harpagoside, and paeoniflorin. The induction cooker was turned on and the solution was slowly heated (to prevent gelatinization and breakdown of the ingredients). The solution was stirred continuously until sufficiently fluid, and the heating power was adjusted to keep the solution warm. The vacuum belt dryer was turned on to dry and powderize the solution. The parameters were as follows: feed rate 1.5 L / h, crawler belt speed 160 mm / min, swing arm angle 26°, cloth motor 100 r / min, and heating temperatures for the first, second, third, and fourth heaters 95±5°C, 95±5°C, 95±5°C, and 30±5°C, respectively. The cutter was turned on for 10 seconds, and the vacuum pressure was 0.0985±0.0015 MPa. The powder was collected in a transport container barrel, sealed, and ready for use. The dried powder was weighed to determine its harpagoside and paeoniflorin content. Approximately 1.5 kg of dry powder was weighed and placed in the hopper of the granulator. The pressure of the pressure roller was set to 10.0 MPa, the screw feeder rotation speed was adjusted to 45 r / min vertically and 120 r / min horizontally, the pressure roller rotation speed was set to 8.0 r / min, and the mesh size of the drying net was set to 16 mesh. During operation, the condition of the equipment and the properties of the granules were observed, and the granulation parameters were adjusted appropriately. Powder was added as needed, and the granules were collected. After screening, the single-step compaction rate and fineness rate were calculated.

[0133] 1.2 Test results Based on the pilot test conditions and detection results of three batches of Liangjitang Granules, mass transfer analysis was carried out for each batch test, and the mass transfer data of the three batches of pilot tests are shown in Table 15 and Figure 10.

[0134] JPEG2025532254000015.jpg56170

[0135] Analysis results: (1) The average extraction yield during the extraction process was 54.72%, the transfer rate of paeoniflorin was 73.08%, and the transfer rate of harpagoside was 68.91%, all of which were equal to or exceeded those of the accompanying decoction (reference sample from the same batch) and the reference sample, indicating that this extraction met the development requirements of the classical prescription. (2) During the processes from extraction to concentration and drying, the dry extraction rate, harpagoside, and paeoniflorin decreased to various degrees, and the component loss rate was greater than the loss in extraction yield. Analysis revealed that losses during the process were mainly concentrated in the vacuum concentration and drying processes. (1) Vacuum concentration process: The losses of paeoniflorin and harpagoside were 16.35% and 6.90%, respectively, greater than the losses in the dry extraction rate. This indicated that the indicator components, especially paeoniflorin, were partially destroyed during the concentration process, which was the main loss step for the indicator components. (2) Drying process: The losses of paeoniflorin and harpagoside were consistent with the dry extraction rate, being 12.52%, 11.49%, and 11.98%, respectively. The losses were mainly due to residues in the container, pipeline, and equipment. (3) Analysis of the main parameters of vacuum concentration, temperature, and time, combined with the study of paeoniflorin stability, revealed that paeoniflorin in Liangji Tang medicinal liquid is susceptible to decomposition when the pH reaches >6 during the vacuum concentration process (temperature 80-90°C). Detection showed that the higher the extract concentration, the lower the pH. The pH of the first and second extracts was low, while the pH of the third extract was higher, >6. Therefore, in the manufacturing process of Liangji Tang granules, it is recommended to concentrate the three extracts in batches and then mix and concentrate again to minimize the heating time for low-concentration paeoniflorin.

[0136] 2. Pilot tests of three batches after optimization (batch concentration + mixed concentration test) (batch numbers 2107015, 2107016, 2107017) 2.1 Test Method 12.309 kg of Rehmannia glutinosa, 12.309 kg of Xanthan Ginseng, 6.138 kg of White Peony Root, 6.138 kg of Wheat Flour, and 3.696 kg of Dioscorea Root were weighed. Seven times the volume of the pieces were added with water and steeped for 40 minutes. The first time, the pieces were heated at 100°C for 30 minutes to extract, the medicinal liquid was released, and then microfiltered. The second time, five times the volume of the pieces were added with water, heated at 100°C for 20 minutes to extract, the medicinal liquid was released, and then microfiltered. The third time, five times the volume of the pieces were added with water, heated at 100°C for 20 minutes to extract, the medicinal liquid was released, and then microfiltered. The volume of each medicinal liquid was measured, and the density, dry extractability, harpagoside, and paeoniflorin were detected. Each drug solution was filtered through a plate and frame filter (membrane pore size: 5-15 μm), and the density of the drug solution was measured (the fine filter plate was replaced when the pressure exceeded 0.02 MPa). The number of times the plate was replaced was recorded. The vacuum was turned on and the first filtrate was sucked into the vacuum concentrator. The steam valve was opened and heating was initiated. The temperature was controlled at 85±5°C, the vacuum at 0.04±0.01 MPa, and the steam pressure at 0.025±0.005 MPa. The drug solution was concentrated to approximately 20 L (density approximately 1.18 g / mL) and then transferred to a liquid storage tank for use. The second filtrate was placed into the vacuum concentrator, the steam valve was opened and heating was initiated. The concentration conditions were the same as the first filtrate. The drug solution was concentrated to approximately 20 L (density approximately 1.10 g / mL) and then transferred to a liquid storage tank for use. The third filtrate was placed into the vacuum concentrator, the steam valve was opened and heating was initiated. The concentration conditions were the same as the first filtrate. The drug solution was concentrated to approximately 20 L (density approximately 1.04 g / mL) and then transferred to a liquid storage tank for use. Weigh out 3.696 kg of gelatinous glue, powderize it, add water three times the mass of gelatinous glue, heat to boiling, and stir constantly to dissolve until a homogeneous gelatinous solution is obtained. Continue concentrating until the density is ≧1.16 g / mL to obtain gelatinous solution. Approximately 2.4 kg of dextrin was weighed and placed in a stainless steel barrel, and water 1 to 2 times the mass of the dextrin was added and stirred until completely dissolved, to obtain a dextrin solution. Turn on the vacuum and place the three concentrated solutions, the glue solution, and the dextrin solution into the vacuum concentration tank. Open the steam valve and control the temperature at 85±5°C, the vacuum at 0.04±0.01MPa, and the steam pressure at 0.025±0.005MPa. Continue concentrating until a thick paste with a density of approximately 1.30g / mL (20°C) is obtained. After filtering through a filter, the paste is placed into a transport container barrel for use. Requirements: The interval between discharging the solution and drying must not exceed 8 hours. The induction cooker was turned on and the solution was slowly heated (to prevent gelatinization and breakdown of the ingredients). The solution was stirred continuously until it was sufficiently fluid, and the heating power was adjusted to keep the solution warm. The vacuum belt dryer was turned on to dry and powderize the solution. The parameters were as follows: feed rate 1.5 L / h, crawler belt speed 160 mm / min, swing arm angle 26°, cloth motor 100 r / min, and heating temperatures for the first, second, third, and fourth heaters 95±5°C, 95±5°C, 95±5°C, and 30±5°C, respectively. The cutter was turned on for 10 seconds, and the vacuum pressure was 0.0985±0.0015 MPa. The powder was collected in a transport container barrel, sealed, and ready for use. Approximately 1.5 kg of dry powder was taken and weighed, and the powder was placed in the hopper. The roller pressure of the granulator was adjusted to 10.0 MPa, and the screw feeder rotation speeds were set to: vertical rotation speed 45 r / min, horizontal rotation speed 120 r / min, roller rotation speed 8.0 r / min, and the drying net mesh number was 16 mesh. The device was then operated. During operation, the state of the device and the properties of the granules were observed (if the granules were loose and finely divided, increase the roller pressure and decrease the feeder rotation speed; if the granules were hard, decrease the roller pressure and decrease the feeder rotation speed). Powder was added as appropriate to collect the granules.

[0137] 2.2 Test results Based on the pilot test conditions and detection results of three batches of Liangji Tang Granules, the dry extraction rate, harpagoside transfer rate, and paeoniflorin transfer rate were used as consideration indicators to analyze the mass transfer conditions during the test. The changes in these consideration indicators during the extraction, batch concentration, mixed concentration, and drying of Liangji Tang Granules were calculated and analyzed. The average mass transfer conditions for the three batches of pilot test are shown in Table 16 (all data are average values ​​for the three pilot test batches), and the loss conditions (loss rate is the amount lost divided by the total extracted amount; all data are average values ​​for the three pilot test batches) are analyzed in Table 17.

[0138] JPEG2025532254000016.jpg73170

[0139] JPEG2025532254000017.jpg51170

[0140] Explanation: (1) The dry extraction rates and paeoniflorin transfer rates of the three pilot batches (2107015, 2107016, 2107017) were basically consistent. The dry extraction rates were 49.66%, 51.43%, and 49.66%, respectively, and the paeoniflorin transfer rates were 60.63%, 63.47%, and 60.00%, respectively. The harpagoside transfer rates of batches 2107015 and 2107017 were consistent at 84.69% and 84.00%, respectively, while batch 2107016 was lower at 57.14%. It was analyzed that this was due to the heterogeneity of Xuanshenan decoction pieces. (2) Analysis of losses of key quality indicators from extract to dry powder in three pilot batches. (1) Dry extraction rate, harpagoside, and paeoniflorin all experienced partial losses (average losses of 21.12%, 17.68%, and 23.67%, respectively). The loss rates were essentially consistent, and analysis revealed that the main cause was material loss, such as pipeline and equipment residues. (2) During the concentration stage, the main quality indicators, dry extraction rate and paeoniflorin losses, were 4.45% and 5.42%, respectively, and the paeoniflorin destruction rate was 1.0%. This demonstrated that individual batch concentration resolved the problem of paeoniflorin destruction during the concentration process. (3) Three batches of pilot tests showed that the average dry extraction rate, harpagoside transfer rate, and paeoniflorin content transfer rate from the decoction pieces to the dry powder were essentially consistent with those of the accompanying decoction (the reference sample from the same batch), at 41.20%, 62.13%, 48.4%, 42.83%, 49.33%, and 48.97%, respectively. This indicates that the pilot test process for Liangji Tang Granules meets the requirements of classical prescriptions (the formulation quality indicators are consistent with the material standards). The optimized bench-scale process for Liangji Tang Granules was verified to be suitable for the production of modern herbal formulations.

[0141] 3. Comparison test results of mixed concentration and batch concentration + mixed concentration (1) Mixing and concentration: After three mixings, the pH of the solution was >6.0, the heating time was 2-4 hours, the concentration of the concentrated solution was low (<0.5 mg / mL), and the destruction rate of paeoniflorin was 18.6%. (2) Batch concentration + mixed concentration: The pH of the first and second extraction solutions was <6.0, and the pH of the third extraction solution was >6.0. The heating time was <2 hours, which was shortened, and the concentration of the concentrate increased (>0.5 mg / mL). The destruction rate of paeoniflorin was 1.2%.

[0142] Based on the principle of "retaining differences while exploring commonalities, respecting the past without clinging to it," this invention examines the formula, origin, baking, decoction process, and medication habits. Through experimental research, the manufacturing process and quality standards for a "Double Decoction Reference Sample" were established, and a "cup of Double Decoction" was solidified and controllable. Modern industrial preparation equipment was used for extraction, purification, concentration, drying, and granulation. Modern analytical detection tools such as HPLC were also used to establish detection methods for multi-component content and characteristic spectra, monitoring the changes in chemical composition before and after each process step in the commercial production process and analyzing the mass transfer state, ensuring the consistency of chemical composition between the industrial preparation and the decoction obtained by traditional techniques. Furthermore, compared with traditional decoctions, the Double Decoction Granules have the advantages of better stability, portability, and ease of administration.

[0143] Through experiments, the present invention determined the amount of decoction pieces and the amount of excipients added to make 1000g of granules in the modern formulation of Liangji Tang, and the formula is: Rehmannia glutinosa 520g, Xanthan Ginseng 520g, Peony Root 260g, Maltodextrin 260g, Dioscorea Root 156g, Euglena Root 156g, and dextrin 100-150g.

[0144] In optimizing the extraction process, the present invention first conducted a preliminary test under pilot production conditions using a reference sample of the two decoctions (using extraction yield, paeoniflorin and harpagoside contents, and characteristic spectra as detection indicators). It was found that the extraction rate (extraction yield, paeoniflorin and harpagoside contents) of the modern formulation was 20-30% higher than that of the accompanying decoction (prepared using the same batch of decoction pieces according to the reference sample manufacturing method). Only after further purification, concentration, drying, and other loss processes could the extraction rate reach the standard of the reference sample. The extraction process for the two decoctions was then screened based on the 20-30% higher extraction rate. The selected extraction process featured an appropriate amount of water, a short time, and high extraction efficiency (high extraction yield, paeoniflorin, and harpagoside). Compared to the over-extraction and high-energy consumption extraction methods used in modern traditional Chinese medicine, which only pursue extraction rates (each extraction usually takes 1-2 hours, and the excess is often impurities such as fiber and starch, which does not increase the extraction of active ingredients and destroys some ingredients even when heated for a long time), this method has improved the quality and medicinal effects of the product and reduced production costs.

[0145] The entire post-extraction process of this invention is carried out at a relatively low temperature (<100°C). The purification process uses physical methods such as centrifugation and plate-frame filtration to remove impurities. The concentration process uses a vacuum concentration method, and the drying process uses vacuum belt drying. The lower temperature helps protect and preserve the ingredients and avoids the destruction of ingredients due to high temperatures. This minimizes process destruction during industrial preparation production and ensures consistency between traditional decoctions and modern preparations.

[0146] The vacuum concentration process employed in this invention differs from traditional Chinese herbal medicines. Research has shown that paeoniflorin is easily destroyed in low-concentration Liangchi Tang medicinal solutions when heated for extended periods. The traditional method for concentrating Chinese herbal medicines involves concentrating the first extract under reduced pressure in a vacuum concentration tank. After the second and third extractions, the extracts are then sequentially concentrated in vacuum concentration tanks to reach a specified density. This method reduces the total concentration of the medicinal solution and extends the absolute heating and concentration time. This invention employs a gradient concentration method using batch individual concentration and blending. The first, second, and third extracts are individually concentrated to a specified density, then blended and concentrated to a specified concentration. This concentration method rapidly increases the paeoniflorin content in the three extracts and shortens the low-concentration concentration time (an environment in which paeoniflorin is easily destroyed). This method finds optimal concentration process conditions for Liangchi Tang granules and solves the problem of the destruction of paeoniflorin, the active ingredient in Liangchi Tang granules. Studies on the stability of paeoniflorin showed that when the paeoniflorin concentration in Liangchi Tang was <0.3 mg / mL and the pH value of the solution was >6.0, paeoniflorin was partially destroyed when concentrated under reduced pressure at a temperature of 80-90°C. However, the paeoniflorin concentration in the third extract of Liangchi Tang Granules was relatively low (<0.04 mg / mL). When the three extracts were mixed and concentrated according to the traditional method of concentrating traditional Chinese medicine, the paeoniflorin concentration in the mixed solution was low and the pH value was high (>6.0). Under the conditions of low concentration and high pH, ​​the paeoniflorin was heated and concentrated for a long time, resulting in partial destruction. After the test, the three extractions of Liangchi Tang Granules were centrifuged and microfiltered, respectively, and the filtrate was sucked into a vacuum concentrator and concentrated to a density of ≥ 1.10 g / mL under reduced pressure conditions (control temperature 80-90°C, vacuum degree 0.03-0.06 MPa, steam pressure 0.03-0.05 MPa). When the density reached ≥ 1.10 g / mL, the paeoniflorin in the concentrated solution was > 0.3 mg / mL, the pH value was < 6.0, and it reached a stable state.

[0147] In the manufacturing process of the Liangchi Tang preparation of the present invention, the excipient dextrin is added during the concentration process, that is, the excipient dextrin is dissolved in water, then sucked into a vacuum concentrator, and mixed with the concentrated medicinal solution and gelatin solution to concentrate. This solves the problems of large air bubbles and material loss caused by sticky bands due to the relatively high viscosity of the Liangchi Tang granule medicinal solution (containing gelatin and polysaccharides) during the vacuum belt drying process. At the same time, compared with the traditional method of adding excipients before granulation, it can solve the problem of uniform mixing between excipients and raw materials in modern granules.

[0148] Because the two decoctions contain heat-sensitive ingredients such as viscous gelatin, polysaccharides, and paeoniflorin, which are prone to aggregation, traditional high-temperature drying and spray drying methods tend to cause aggregation, destroying heat-sensitive active ingredients such as paeoniflorin. The drying method used in this invention is vacuum belt drying, which has the advantages of low drying temperature, short drying time, minimal loss, and high efficiency. The resulting two decoctions do not aggregate during the drying process, and heat-sensitive ingredients such as paeoniflorin are not destroyed. The two decoctions have uniformly distributed ingredients, a high content of active ingredients such as paeoniflorin, and good medicinal efficacy.

[0149] The quality standards for the Liangji Tang formulation provided by the present invention specify upper and lower limits for harpagoside and paeoniflorin. After completing research on the manufacturing process of the reference sample, a comprehensive quality control mode based on the reference sample was established. The overall chemical composition of the reference sample was comprehensively monitored using index component content + characteristic spectrum technology. The established multi-component control index was used to conduct product stability studies to ensure the marketability of the finished drug. "Liangji Tang" is a famous ancient classic formula. Using modern pharmaceutical equipment and technology, the modern formulation of the present invention is obtained by optimizing the extraction, concentration, drying, and granulation processes. At the same time, a quality control system was established for the entire process of "medicinal material-drinking pieces-preparation" to ensure that the resulting formulation maintains the consistency and stability of traditional decoctions in terms of their material basis. It also possesses the advantages of modern formulations, such as stable quality, portability, and ease of administration.

[0150] The above is only a preferred embodiment of the present invention. It should be pointed out that those skilled in the art can make some improvements and modifications without departing from the principle of the present invention, and these improvements and modifications shall also fall within the protection scope of the present invention.

Claims

1. A method for producing a water extract of both local decoctions, comprising: mixing five kinds of Chinese herbal medicines with first water, and sequentially carrying out first steeping and first extraction to obtain a first extract and a first medicine residue; the five kinds of Chinese herbal medicines are Rehmannia Root, Xanthan Ginseng, Peony Root, Wheat Flour, and Dioscorea Ricinus; Mixing the first drug residue with second water, and sequentially performing second immersion and second extraction to obtain a second extraction liquid and a second drug residue; Mixing the second drug residue with third water, and sequentially performing a third soaking and a third extraction to obtain a third extract; concentrating the first extract, the second extract, and the third extract to obtain a first concentrate, a second concentrate, and a third concentrate, respectively; wherein the concentration temperature is independently 70 to 90°C, the vacuum degree is independently 0.03 to 0.07 MPa, and the steam pressure is independently ≦0.09 MPa; and mixing the first concentrated solution, the second concentrated solution, the third concentrated solution and the gelatin solution to obtain a water extract of both the tea tangs, The mass ratio of the Rehmannia glutinosa root, Xanthan Ginseng root, Peony root root, Dioscorea officinalis root, and Euglena pith is 37.30:37.30:18.65:18.65:11.19:11.

19.

2. The mass of the first water is 6 to 8 times the mass of the five kinds of Chinese herbal medicines; The mass of the second water and the third water is independently 4 to 6 times the mass of the five kinds of Chinese herbal medicines; the temperatures of the first extraction, the second extraction, and the third extraction are independently 98 to 101°C; the first steeping and first extraction times are independently between 30 and 50 minutes; 2. The method of claim 1, wherein the second steeping, second extraction, third steeping and third extraction times are independently 20 to 40 minutes.

3. The manufacturing method described in claim 1 or 2, characterized in that after the first extraction, the obtained first extraction system is also post-treated, and the post-treatment includes centrifuging the obtained first extraction system to obtain a liquid component and a first drug residue, and microfiltrating the liquid component to obtain a first extract.

4. 4. The method according to claim 3, wherein the microfiltration is performed using a plate and frame filter, and the pore size of the filter membrane of the plate and frame filter is 5 to 15 μm.

5. 2. The method of claim 1, wherein the densities of the first concentrate, the second concentrate, and the third concentrate are independently 1.10 g / mL or more and less than 1.26 g / mL.

6. 2. The method for producing the gelatinous liquid according to claim 1, comprising the steps of dissolving the gelatinous liquid in water and then sieving the solution, the underside of the sieve being the gelatinous liquid, the mass of the water being 2 to 6 times the mass of the gelatinous liquid, and the sieve size of the sieve being 100 mesh.

7. A water extract of both local tea leaves produced by the method according to any one of claims 1 to 6.

8. A double-water extract paste, obtained by concentrating the double-water extract liquid described in claim 7, characterized in that the density of the double-water extract paste is 1.2 to 1.3 g / mL.

9. A dual-tang preparation, the manufacturing raw materials of which include a dual-tang aqueous extract and a pharmaceutically acceptable excipient, the dual-tang aqueous extract being the dual-tang aqueous extract liquid described in claim 7 and / or the dual-tang aqueous extract ointment described in claim 8, the mass ratio of the pharmaceutically acceptable excipient to the dual-tang aqueous extract is 1: (12.48-18.72), and the mass of the dual-tang aqueous extract is calculated based on the total mass of Rehmannia Root, Xanthan Ginseng, Peony Root, Magnolia Officinalis, Rehmannia Root and Erjiao.

10. The two-way decoction preparation according to claim 9, characterized in that, when calculated by parts by mass, the raw materials for producing 1,000 parts of the two-way decoction preparation include 468-572 parts of Rehmannia glutinosa, 468-572 parts of Xanthan Gum, 234-286 parts of Peony Root, 234-286 parts of Wheat Flour, 140-171 parts of Dioscorea Root, 140-171 parts of Egg Gluconate, and 100-150 parts of a pharmaceutically acceptable excipient.

11. The preparation according to claim 9 or 10, characterized in that the water content of the preparation is 3 to 8 wt %.

12. The two-decoction preparation according to claim 9 or 10, characterized in that the two-decoction preparation contains two-decoction powder or two-decoction granules, the particle size of the two-decoction powder being 150 to 850 μm, and the particle size of the two-decoction granules being 180 to 2000 μm.

13. mixing an aqueous solution of the two geotang extract and a pharmaceutically acceptable excipient, and concentrating the mixture to obtain a concentrated mixture; The method for producing a Ryotang preparation according to any one of claims 9 to 12, characterized in that it comprises a step of vacuum belt drying the mixed concentrated liquid to obtain a Ryotang preparation.

14. 14. The method according to claim 13, wherein the aqueous solution of the pharmaceutically acceptable excipient is obtained by dissolving the pharmaceutically acceptable excipient in water, and the mass ratio of the pharmaceutically acceptable excipient to water is 1:(2-6).

15. The method according to claim 13, wherein the concentration temperature is 70 to 90° C., the vacuum degree is 0.03 to 0.07 MPa, and the steam pressure is ≦0.09 MPa.

16. The method according to claim 13 or 15, wherein the density of the mixed concentrate is 1.25 to 1.36 g / mL.

17. 14. The method of claim 13, wherein the vacuum belt drying is performed using a vacuum belt dryer, and the vacuum belt drying parameters include a supply temperature of 75-85°C, a supply rate of 6-24 L / h, a crawler belt running speed of 16±5 cm / min, a swing arm angle of 65-85°C, a fabric motor speed of 30-50 r / min, heating temperatures of zone 1, zone 2, zone 3 and zone 4 of 95±5°C, 96±5°C, 96±5°C and 30±5°C, respectively, a cutter speed of 10-20 seconds / cycle and a vacuum intensity of 0.097-0.1 MPa.

18. When the two-way decoction preparation is two-way decoction granules, the two-way decoction granules are obtained by further granulating the two-way decoction granules after the vacuum belt drying. The method according to claim 13 or 17, wherein the granulation is carried out using a granulator, and the granulation parameters include a roller pressure of the granulator of 12 to 20 MPa, a vertical rotation speed of the screw feeder of 18 to 30 r / min, a horizontal rotation speed of the screw feeder of 90 to 140 r / min, a roller rotation speed of 4 to 10 r / min, and a sizing speed of 100 to 150 r / min.

19. A quality control standard for the two-tang preparation according to any one of claims 9 to 12 or the two-tang preparation produced by the production method according to any one of claims 13 to 18, comprising the HPLC characteristic spectrum, characteristic component content, water content, and dry extraction rate of the two-tang preparation; The content of the characteristic components includes 0.33 to 0.67 mg / g of harpagoside, 2.62 to 4.96 mg / g of paeoniflorin, 8.83 to 16.42 mg / g of L-hydroxyproline, and 17.42 to 32.46 mg / g of glycine. The water content is 3 to 8 wt %, A quality control standard characterized in that the dry extraction rate is 40-55%.

20. The method for obtaining the HPLC characteristic spectra of both the above-mentioned Jingtang preparations is as follows: Step (1): Mixing and concentrating the test subject Ryojito preparation, diatomaceous earth, and acetonitrile aqueous solution, and dissolving the resulting concentrate in a methanol aqueous solution to obtain a test substance solution; Step (2) mixing paeoniflorin control substance, harpagoside control substance and methanol to obtain a control substance solution; (3) subjecting the test substance solution and the control substance solution to HPLC detection, respectively, to obtain the control substance characteristic spectrum and the test substance characteristic spectrum; and calculating the relative retention times of other characteristic peaks in the test substance characteristic spectrum based on the retention time of the paeoniflorin control substance in the control substance characteristic spectrum, thereby obtaining the HPLC characteristic spectra of both Reitang preparations; 20. The quality control method of claim 19, wherein steps (1) and (2) have no chronological order.

21. The conditions for the HPLC detection were as follows: a chromatography column using octadecylsilane-bonded silica as a packing material; mobile phase A being methanol; mobile phase B being 0.05 v / v% phosphoric acid solution; a mobile phase flow rate of 0.8 to 1.2 mL / min; and gradient elution, the gradient elution procedure being as follows: 【Table 1】 The quality control standard of claim 20, comprising a detection wavelength of 254 nm, a column temperature of 28°C, and a sample load volume of 10 μL.

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