Preparation method of Jindan attached extension particles

Abstract

The invention provides a preparation method of Jindan attached extension granules, and belongs to the technical field of traditional Chinese medicine preparations. According to the preparation method disclosed by the invention, an existing preparation process of the Jindan attached-extension granules is further improved and optimized, after a volatile oil inclusion compound of beta-cyclodextrin is formed, gelled silicon dioxide is added, and the gelled silicon dioxide is adhered to the beta-cyclodextrin by adopting high-substituted hydroxy propyl cellulose to form a composite inclusion. Through optimization and combination of various methods such as optimization of the variety and the addition amount of the gelled silicon dioxide, the addition amount, the variety and the solvent of the hydroxy propyl cellulose, nitrogen filling and the like, overflowing volatile components can be fully adsorbed in the gelled silicon dioxide, so that the stability of the volatile components in the Jindan attached-extension particles is greatly improved, and the stability of the volatile components in the Jindan attached-extension particles is improved. And the loss of volatile oil in the shelf life of the product is obviously reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a method for preparing Jindan Fuyan granules. Background Art

[0002] Jindan Fuyan Granules are Chinese medicine granules with national medicine standard Z20120018. They are made of 13 herbs: Peony Bark, Honeysuckle, Red Peony Root, Curcuma Atractylodes, Vinegar Corydalis, Patrinia, Sargentodoxae, Peach Kernel, Angelica Sinensis, Vinegar Myrrh, Cinnamon Twig, Cyperus Rhizome, and Coix Seed. The medicine has the effects of clearing away heat and dampness, promoting blood circulation and removing blood stasis, promoting qi and relieving pain. It can be used for chronic pelvic inflammatory disease, and has good clinical effects for patients with internal dampness and heat according to Chinese medicine dialectics. There are many herbs containing volatile components in the prescription, and the production process is complicated. The current production process uses steam distillation to collect volatile oils from herbs such as Cinnamon Twig, Curcuma Atractylodes, Myrrh, Cyperus Rhizome, and Angelica Sinensis. Modern pharmacological studies have shown that volatile oil components play an important anti-inflammatory and analgesic role in the treatment of chronic pelvic inflammatory disease with Jindan Fuyan Granules.

[0003] β-cyclodextrin is a special structure of a hollow ring-shaped cylinder, which is formed by enzymatic cyclization of starch. Its inner cavity provides a space suitable for oil molecules to enter. Volatile components, as a kind of substances that are more volatile at room temperature, can be encapsulated in the ring structure of β-cyclodextrin through inclusion with β-cyclodextrin to form ultrafine particle dispersions. This inclusion mechanism not only improves the solubility and stability of volatile oils, but also effectively reduces the volatilization of volatile components. However, through research, it was found that the content of volatile components in Jindan Fuyan granules prepared by β-cyclodextrin inclusion will gradually decrease over time during the shelf life, but the speed and magnitude of the decrease are lower than those of products produced without inclusion technology. The reason is that some volatile components may overflow from the ring structure of β-cyclodextrin, be adsorbed on the packaging material or pass through the packaging material, and be oxidized by air and cause losses. Therefore, in order to improve the stability of volatile oil components and ensure the efficacy of Jindan Fuyan granules, it is necessary to further study and improve its preparation process. Summary of the invention

[0004] In view of the background technology, the purpose of the present invention is to provide a method for preparing Jindan Fuyan granules. In order to further improve the stability of volatile oil in Jindan Fuyan granules, the present invention adopts a new process of β-cyclodextrin inclusion, gelled silica adsorption, hydroxypropyl cellulose adhesion combination to form inclusions and nitrogen filling, which ultimately makes the volatile components of Jindan Fuyan granules more stable during the shelf life and reduces the loss rate from 23.03% to 1.90%. The effect of the new preparation process on the efficacy of the product is further verified through animal experiments.

[0005] To achieve the above purpose, the present invention specifically adopts the following technical solutions: A method for preparing Jindan Fuyan granules comprises the following steps: Step 1: Take the vinegar and alcohol extract of Corydalis yanhusuo, and concentrate the extract into a thick paste for later use; Step 2: extract paeonol from peony bark by steam distillation, and then encapsulate it with β-cyclodextrin to obtain paeonol inclusion complex; collect the water after distillation in another device; Step 3: extract volatile oil from Curcuma zedoaria, Angelica sinensis, Myrrha acetophylla, Cinnamomum cassiae and Cyperus rotundus by steam distillation, then include it with β-cyclodextrin to obtain volatile oil inclusion compound, add gelled silicon dioxide to the volatile oil inclusion compound and mix well, then continue to add hydroxypropyl cellulose and mix well to obtain volatile oil inclusion body; collect the water liquid after distillation in another device; Step 4: Mix the residues after extraction in steps 2 and 3 with honeysuckle, red peony root, Patrinia chinensis, Sargentodoxae, peach kernel, and coix seed, add water and boil, combine the filtrate after filtering with the water solution after distillation in steps 2 and 3, concentrate into a clear paste, precipitate with alcohol, concentrate and combine with the thick paste obtained in step 1, add sucrose and dextrin, and finally add the paeonol inclusion compound obtained in step 2 and the volatile oil inclusion body obtained in step 3, mix well, and granulate to obtain the product.

[0006] Furthermore, the raw materials of the Jindan Fuyan granules are as follows by weight: 260 parts of peony bark, 260 parts of honeysuckle, 220 parts of red peony root, 160 parts of Curcuma, 200 parts of vinegar Corydalis, 200 parts of Patrinia, 200 parts of Sargentodoxa, 160 parts of peach kernel, 160 parts of Angelica, 160 parts of vinegar Myrrh, 160 parts of cinnamon twig, 160 parts of Cyperus, and 260 parts of Coix seeds.

[0007] Furthermore, the colloidal silica in step 3 is SYLOID ® XDP、SYLOID ® 244P or SYLOID ® At least one of 63FP, preferably SYLOID ® XDP.

[0008] Furthermore, in step 3, the mass ratio of the volatile oil inclusion compound to the colloidal silica is (5-20):1, preferably 10:1.

[0009] Furthermore, the viscosity of the hydroxypropyl cellulose in step 3 is 50-4000 mPa·s, and the molar substitution degree is 3.5-4.5; preferably, the viscosity is 150 mPa·s.

[0010] Furthermore, in step 3, the hydroxypropyl cellulose is added after being prepared into a solution, the concentration of which is 3%-10%, and the solvent used is ethanol, water or a mixture of the two; preferably, the concentration of the solution is 5%, and the solvent is a 50% by volume ethanol aqueous solution.

[0011] Furthermore, the weight volume ratio of the mixture obtained by mixing the volatile oil inclusion compound and the gelled silicon dioxide to the hydroxypropyl cellulose solution is 2:1 (kg / L).

[0012] Furthermore, step four also includes packaging the granulated product and filling it with nitrogen.

[0013] β-cyclodextrin is an odorless, non-toxic white crystalline powder, a ring-structured compound composed of 7 pyranose glucose molecules, with a conical cylindrical molecular structure that is hydrophobic inside and hydrophilic outside, which can change the physical and chemical properties of the guest molecule, such as solubility, volatility and chemical properties. Colloidal silica is a fine powder particle, widely used in oral and some topical preparations. It is a non-toxic and non-irritating pharmaceutical excipient with small particle size, large surface area, strong adsorption capacity and good biocompatibility. It can fix volatile components in its internal high-porosity three-dimensional network structure through physical adsorption, thereby reducing the volatility of volatile oils. β-cyclodextrin uses its internal hydrophobic and external hydrophilic properties to enclose volatile oil molecules in its cavity; while colloidal silica uses its excellent adsorption and dispersibility to further adsorb and fix the volatile oil molecules escaping from the inclusion compound and reduce its contact area with air. Both can adsorb volatile oil, but the loss rate of volatile components during the shelf life is still high when volatile oil is adsorbed only by β-cyclodextrin inclusion or gelled silica. The present invention uses β-cyclodextrin to encapsulate volatile oil, and then uses hydroxypropyl cellulose to adhere gelled silica to β-cyclodextrin to form a composite inclusion body, which can more effectively reduce the volatility of volatile oil and improve the dispersibility of volatile oil in water compared with using β-cyclodextrin inclusion or gelled silica adsorption alone, and can be used to improve the stability of volatile components in Jindan Fuyan granules, extend the shelf life or enhance the therapeutic effect.

[0014] Compared with the prior art, the present invention has the following beneficial effects: The present invention improves the original preparation process of Jindan Fuyan granules, and adds gelled silicon dioxide and hydroxypropyl cellulose on the basis of β-cyclodextrin encapsulation, and adheres the gelled silicon dioxide to β-cyclodextrin through hydroxypropyl cellulose to form a composite inclusion body; compared with the original preparation process, the stability of volatile oil is significantly improved, and the loss of volatile oil in the shelf life of Jindan Fuyan granules is reduced. According to the rat chronic pelvic inflammatory disease model experiment, it is also proved that the Jindan Fuyan granules prepared by the improved process of the present invention have significant advantages in efficacy compared with the original process. DETAILED DESCRIPTION

[0015] The technical solution of the present application will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present application and its application or use. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present application.

[0016] Example The original preparation process of Jindan Fuyan Granules is: 1. According to the prescription, weigh 260 g of Peony Bark, 260 g of Honeysuckle, 220 g of Red Peony Root, 160 g of Curcuma, 200 g of Vinegar Corydalis, 200 g of Patrinia, 200 g of Sargentodoxa, 160 g of Peach Kernel, 160 g of Angelica, 160 g of Vinegar Myrrh, 160 g of Cinnamon Twig, 160 g of Cyperus, and 260 g of Coix Seed for later use.

[0017] 2. For the above 13 Chinese medicinal materials, vinegar Corydalis yanhusuo was crushed into coarse powder and extracted three times with 60% (volume fraction, the same below) ethanol: for the first time, 8 times the mass of 60% ethanol was added, soaked for 24 hours, heated under reflux for 2 hours, and filtered; for the second time, 6 times the mass of 60% ethanol was added, heated under reflux for 2 hours, and filtered; for the third time, 6 times the mass of 60% ethanol was added, heated under reflux for 1 hour, and filtered; the three extracts were combined, filtered, ethanol was recovered under reduced pressure, and concentrated to a thick paste with a relative density of 1.32 (60°C) for use. Add 6 times the mass of water to the bark of peony, use steam distillation to extract paeonol for about 6 hours, collect the distillate, and after the distillate is cooled to room temperature, put it into a cold storage room at 2-10°C for refrigeration and brain separation, collect the precipitated paeonol, and collect the water liquid after distillation in another container; dry the paeonol at 40°C, add an appropriate amount of anhydrous ethanol to dissolve it, take β-cyclodextrin with 12 times the mass of paeonol, grind the β-cyclodextrin with 2 times the mass of water, add the ethanol solution of paeonol dropwise, grind it sufficiently to obtain the paeonol inclusion compound, dry it at 40°C, crush it and set aside. Take five Chinese medicinal herbs including cinnamon twig, curcuma zedoaria, vinegar myrrh, cyperus rotundus and angelica sinensis, add 5 times the weight of water, extract volatile oil by steam distillation for about 6 hours, collect the volatile oil, and collect the water liquid after distillation in another container; take β-cyclodextrin with 9 times the weight of the volatile oil, add water at a ratio of β-cyclodextrin / water = 1:20 (W / V, kg / L), slightly heat to dissolve, then slowly add dropwise the volatile oil under stirring, continue stirring at 30°C for 3 hours, refrigerate (0-4°C) and stand for 24 hours, filter with suction, dry at 40°C, and grind to obtain volatile oil inclusion compound for later use.

[0018] 3. Add water to the residues after extracting the peony bark and five Chinese medicinal materials and the remaining six Chinese medicinal materials, honeysuckle, red peony root, peach kernel, Patrinia scabra, Sargentodoxae, and Coix seeds, and decoct them twice. Add 10 times the mass of water for the first time and 8 times the mass of water for the second time. Each time for 2 hours, combine the decoctions, filter, combine the filtrate with the above distilled water, concentrate to a clear paste with a relative density of 1.09 (60°C), add an equal amount of 95% ethanol to precipitate, let stand for 48 hours, take the supernatant, reduce pressure to recover ethanol and concentrate to a thick paste with a relative density of 1.32 (60°C), combine with the above thick paste, add appropriate amount of sucrose and dextrin, dry under reduced pressure at 60°C, crush into fine powder, make granules, dry, add the above paeonol and volatile oil inclusion compound to mix well, add appropriate amount of starch and dextrin to granulate (1000 g), and pack into small bags (15 g / bag) to obtain (code SH).

[0019] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® XDP (Grace Company, USA) was added and mixed evenly, and the rest were granulated according to the original preparation process of Jindan Fuyan Granules and packaged into small bags (No. SH1).

[0020] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well, and the rest are granulated according to the original preparation process of Jindan Fuyan Granules, and packaged into small bags (No. SH2).

[0021] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well, and the rest are granulated according to the original preparation process of Jindan Fuyan Granules, and packaged into small bags (No. SH3).

[0022] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® 244 P (Grace Company, USA), mix well, and the rest are granulated according to the original preparation process of Jindan Fuyan Granules, and packaged into small bags to obtain (No. SH4).

[0023] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After adding 63 FP (Grace Company, USA), mix well, and the rest are granulated according to the original preparation process of Jindan Fuyan Granules, and packaged into small bags (No. SH5).

[0024] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well; then add 3% hydroxypropyl cellulose (150 mPa·s; molar substitution degree 4, the same below) in 50% ethanol solution at a weight-to-volume ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules and packaged into small bags to obtain (No. SH6).

[0025] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well; then add 5% hydroxypropyl cellulose (150 mPa·s) in 50% ethanol solution at a weight-to-volume ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules and packaged into small bags to obtain (No. SH7).

[0026] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well; then add 10% hydroxypropyl cellulose (150mPa·s) in 50% ethanol solution at a weight-to-volume ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules and packaged into small bags to obtain (No. SH8).

[0027] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well; then add a 5% hydroxypropyl cellulose (150 mPa·s) aqueous solution at a weight-to-volume ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules and packaged into small bags to obtain (No. SH9).

[0028] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well; then add 5% hydroxypropyl cellulose (4000mPa·s) in 50% ethanol solution at a weight volume ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules and packaged into small bags to obtain (No. SH10).

[0029] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID® After XDP, mix well; then add pregelatinized starch slurry (prepared by hot-melt method, concentration of 10%) at a mass ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules and packaged into small bags (No. SH11).

[0030] According to the original process, cinnamon twig, zedoary turmeric, myrrh, cyperus rotundus, and angelica were steam distilled and the volatile oil was collected. Then SYLOID was directly added at a volume-weight ratio of 1:1. ® XDP are mixed, and the obtained substance is used to replace the volatile oil inclusion compound. The rest are granulated according to the original preparation process of Jindan Fuyan granules and packaged into small bags to obtain (No. SH12).

[0031] Jindan Fuyan granules were produced according to the original process mentioned above, and nitrogen was filled into small packages (No. SN1).

[0032] According to the original process, cinnamon twig, zedoary zedoary, myrrh, cyperus rotundus and angelica were used to prepare volatile oil inclusion complexes, and SYLOID ® After XDP, mix well; then add 5% hydroxypropyl cellulose (150 mPa·s) in 50% ethanol solution at a weight-to-volume ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules, packaged into small bags and filled with nitrogen to obtain (No. SN2).

[0033] Test example Stability study: Using the temperature and humidity of accelerated stability study, all samples were placed in a constant temperature and humidity chamber (ICH260 type, Memmert, Germany) at a temperature of 40℃±2℃ and a relative humidity of 75%±5% for 6 months (equivalent to 2 years of long-term stability test). The content of α-cyperone in volatile oil was selected as the object of study, and the scheme was optimized.

[0034] The reason for selecting α-cyperone as the evaluation index is that α-cyperone is an oily yellow liquid, and the volatile oil is extracted from Cyperus rotundus by steam distillation. It is volatile at room temperature and has antipyretic, analgesic and anti-inflammatory effects. α-cyperone has a significant inhibitory effect on Staphylococcus aureus, Pseudomonas aeruginosa and some fungi, and is one of the many effective volatile components mentioned.

[0035] Determination method of α-cyperone: Instruments and equipment: Agilent 1260 Infinity Ⅱ high performance liquid chromatograph (Agilent Technologies, USA); Shimadzu LC-20AT high performance liquid chromatograph (Shimadzu Corporation, Japan); Sartorius Secura125-1CN electronic balance (Sartorius Scientific Instruments Co., Ltd.); Mettler Toledo ML204 electronic balance (Mettler-Toledo Instrument Co., Ltd.).

[0036] Chromatographic column: Agilent 5 HC C18 column (250 mm×4.6 mm, 5 μm); mobile phase: acetonitrile-0.1% phosphoric acid solution (70:30); detection wavelength is 290 nm.

[0037] Preparation of reference solution: Take an appropriate amount of α-cyperone reference substance, weigh it accurately, and add methanol to make a solution containing 50 μg of α-cyperone per 1 mL.

[0038] Preparation of test solution: Take the product under the item of difference in filling amount, grind it into powder, take 5 g, weigh it accurately, put it in a stoppered conical flask, add 50 mL of methanol accurately, weigh it, treat it ultrasonically (power 500 W, frequency 40 kHz) for 45 minutes, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain it.

[0039] Determination method: Accurately pipette 10 μL of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0040] Calculate the amount of α-cyperone (C 15 H 22 The results are shown in Table 1.

[0041] Table 1 α-Cyperone content

[0042] Note: Loss rate (%) = (initial content per bag - content per bag measured after 6 months) / initial content per bag × 100%.

[0043] From the above data, it can be seen that the sample numbered SH produced according to the original process, after 6 months of accelerated stability study, the α-cyperone content was reduced by 23.03%.

[0044] SH1, SH2, and SH3 samples were added with SYLOID at a mass ratio of 20:1, 10:1, and 5:1, respectively. ®As for the samples of XDP colloidal silica, the data show that during the investigation period, the α-cyperone content of SH1, SH2 and SH3 samples decreased by 17.14%, 11.13% and 11.01% respectively; this shows that the addition of 20:1, although the loss rate is smaller than that of the samples without colloidal silica, is worse than that of 10:1 and 5:1, and there is no significant difference between 10:1 and 5:1, indicating that after the addition reaches 10:1, the addition of SYLOID ® XDP colloidal silica has no further effect. For cost considerations, it is best to add it in a mass ratio of 10:1.

[0045] Samples SH4 and SH5 were added with 244P and 63FP colloidal silica at a mass ratio of 10:1. During the observation period, their α-cyperone content decreased by 14.13% and 15.34%, respectively. Compared with the SH2 sample added with XDP at a mass ratio of 10:1, the reduction in α-cyperone was quite different. Therefore, it is better to use XDP colloidal silica. XDP colloidal silica performs well in converting liquids into free-flowing solids due to its optimized mesoporous structure and efficient liquid loading capacity, and is suitable for the dispersion and adsorption of oily active ingredients and lipid systems.

[0046] SH6, SH7, and SH8 are β-cyclodextrin silica inclusions prepared by adding 3%, 5%, and 10% low-viscosity high-substituted hydroxypropyl cellulose (150 mPa·s) 50% ethanol solution at a weight-volume ratio of 2:1 after adding XDP colloidal silica; SH9 is a β-cyclodextrin silica inclusion prepared by adding 5% low-viscosity high-substituted hydroxypropyl cellulose (150 mPa·s) aqueous solution; SH10 is a β-cyclodextrin silica inclusion prepared by adding 5% medium-viscosity high-substituted hydroxypropyl cellulose (4000 mPa·s) 50% ethanol solution. From the data, the α-cyperone content of SH6, SH7, SH8, SH9, and SH10 decreased by 8.22%, 5.31%, 9.37%, 14.38%, and 12.27% respectively during the observation period. The results showed that SH7 used 5% low-viscosity high-substituted hydroxypropyl cellulose in 50% ethanol solution as a binder, which had just the right viscosity and could form stable β-cyclodextrin-silica inclusions. SH6 used 3% low-viscosity high-substituted hydroxypropyl cellulose in 50% ethanol solution, which could not effectively bond silica to the surface of β-cyclodextrin. SH8 used 10% low-viscosity high-substituted hydroxypropyl cellulose in 50% ethanol solution, which had too high a concentration. While adhering, a film was formed, which blocked the pores of silica and failed to achieve good adsorption. As a result, the volatile components overflowing from β-cyclodextrin could not be well adsorbed by the gelled silica. SH9 used 5% low-viscosity high-substituted hydroxypropyl cellulose aqueous solution as a binder. Due to the fact that β-cyclodextrin is slightly soluble in water, the structure of β-cyclodextrin adsorbing volatile oil would be unstable, resulting in poor volatile oil encapsulation effect, and ultimately an increase in the loss rate of volatile components. SH10 uses 5% medium-viscosity high-substituted hydroxypropyl cellulose in 50% ethanol solution as a binder, which is not as effective as using low-viscosity substituted hydroxypropyl cellulose. The possible reason is that the medium-viscosity high-substituted hydroxypropyl cellulose is too viscous and forms a film on the surface of gelled silica, resulting in a decrease in adsorption. SH11 uses pregelatinized starch slurry as a binder, which is too viscous and will form a hard film that blocks the pores of silica, failing to achieve the expected adsorption effect.

[0047] At the same time, the sample after adding 5% low-viscosity high-substituted hydroxypropyl cellulose solution to adhere silica to β-cyclodextrin (SH7, loss rate: 5.31%) is much better than the sample directly mixed with XDP gelled silica in the volatile oil inclusion complex (SH2, loss rate: 11.13%). Considering that the direct addition of silica, the inclusion of silica and β-cyclodextrin is not tight, not as close as the composite inclusion, and cannot effectively absorb the overflowed volatile components.

[0048] SH12 is a sample prepared by only using colloidal silica to adsorb volatile oil. Compared with the original process sample SH, which only used β-cyclodextrin inclusion, the α-cyperone content of SH12 and SH decreased by 10.22% and 23.03% respectively during the accelerated stability test. ® XDP colloidal silica disperses and adsorbs volatile oils, and there will also be a loss of volatile components during the stability study period.

[0049] SN1 was produced according to the original process, and the small package was filled with nitrogen to isolate the sample from oxygen. The results showed that after 6 months of stability study, the α-cyperone content of SH and SN1 samples decreased by 23.03% and 19.21%, respectively. This shows that filling with nitrogen can prevent the oxidation of volatile components in volatile oils and further improve the stability of volatile oils.

[0050] The final optimal production process is: after preparing the volatile oil inclusion compound according to the original process, SYLOID ® After XDP, mix well; then add 5% hydroxypropyl cellulose (E50) in 50% ethanol solution at a weight volume ratio of 2:1, mix well, dry at 40°C, and crush. The rest are granulated according to the original preparation process of Jindan Fuyan Granules, packaged into small bags and filled with nitrogen (SN2).

[0051] Compared with the original process of only using β-cyclodextrin for inclusion, the process of the present invention adds gelled silicon dioxide after forming the volatile oil inclusion complex of β-cyclodextrin, and uses low-viscosity high-substituted hydroxypropyl cellulose to adhere the gelled silicon dioxide to β-cyclodextrin to form a composite inclusion. By optimizing the type and amount of gelled silicon dioxide, the amount, type and solvent of hydroxypropyl cellulose, and the combination of nitrogen filling and other methods, the overflowed volatile components can be fully adsorbed in the gelled silicon dioxide, so that the loss rate of α-cyperone, a representative component in volatile oil, is reduced from 23.03% to 1.90% in the 6-month accelerated stability study, which is significantly superior to the original process.

[0052] Pharmacodynamic evaluation: SPF grade SD female rats (Hunan Slake Jingda Experimental Animal Co., Ltd.), weighing 250 g ± 20 g; mixed bacterial solution (Wuxi Saiwei Technology Co., Ltd.); Rat IL-6 ELISA kit, Rat IL-8 ELISA kit, Rat IL-1β ELISA kit, Rat TNF-α ELISA kit (all purchased from Jiangsu Enzyme Immunity Industry Co., Ltd.); MULTISKAN GO microplate reader (Thermo Fisher Scientific Inc., USA); pipette (Thermo Fisher Scientific Inc., USA); ST16R low temperature centrifuge (Thermo Fisher Scientific Inc., USA). Sample 1: Jindan Fuyan granules (sample SH), sample 2: Jindan Fuyan granules (sample SN2), samples 1 and 2 were placed in a constant temperature and humidity chamber (German Memeltech ICH260 model) at a temperature of 40℃±2℃ and a relative humidity of 75%±5% for 6 months, and pharmacodynamic experiments were carried out to investigate the pharmacodynamic differences between samples 1 and 2 after 6 months of accelerated stability conditions.

[0053] Establishment of rat chronic pelvic inflammatory disease model: Bacterial solution configuration: Staphylococcus aureus, Escherichia coli, and beta-hemolytic Streptococcus were mixed in a ratio of 1:2:1 and prepared with NaCl to a final concentration of 2.0×10 9 CFU / mL bacterial solution mixture. The rats were fixed, weighed, and anesthetized with 3% sodium pentobarbital injection (dose, 30 mg / kg) intraperitoneally. After disinfection with iodine, a 1.5 cm incision was made along the midline of the abdomen under sterile conditions. The left uterus of the rat was fixed and exposed, the left endometrial tissue was mechanically damaged, and 0.1 mL of mixed bacterial solution was slowly injected into the direction of the ovary. The right uterus was not scratched and not injected with bacterial solution as a self-control. The uterus was returned to the abdominal cavity to establish a chronic pelvic inflammatory disease model, the abdomen was closed in layers, and the incision was sutured. After the modeling was completed, each group resumed drinking water, cleaned normally, and raised in separate cages.

[0054] Experimental groups: 1. Blank group: 8 mice, no special treatment.

[0055] 2. Model group: 8 rats, no special treatment after modeling.

[0056] 3. Drug group 1: 8 rats. After modeling, 4.72 g / kg Jindan Fuyan Granules (SH) sample (dissolved in water and administered by gavage) was administered according to the body surface area conversion method between humans and rats.

[0057] 4. Drug group 2: 8 rats. After modeling, 4.72 g / kg Jindan Fuyan Granules (SN2) sample (dissolved in water and administered by gavage) was administered according to the body surface area conversion method between humans and rats.

[0058] The above four groups were intervened for 30 consecutive days.

[0059] Determination of inflammatory factors in rats: On the 31st day after administration, animals were anesthetized by intraperitoneal injection of sodium pentobarbital, and about 4-5 mL of abdominal aortic blood was drawn and placed in a test tube without anticoagulant. After standing at room temperature for 4 hours, the blood was centrifuged at 3000 r / min in a low-temperature high-speed centrifuge for 5 minutes to separate the serum. Take IL-6, IL-8, IL-1β, and TNF-α enzyme-labeled plates, set standard wells, test sample wells, and blank control wells, and record the position of each well. Add 50 μL of standard working solution of different concentrations to the standard wells; draw 10 μL of each serum sample to be tested in the sample wells, and then add 40 μL of sample diluent, and do not add blank control wells. Add 100 μL of horseradish peroxidase (HRP)-labeled detection antibody to each well except the blank well, seal the reaction well with a sealing film, incubate in a 37℃ water bath for 60 minutes, discard the liquid in the well, wash the plate repeatedly, add 50 μL of substrate A and B, and incubate at 37℃ in the dark for 15 minutes. 50 μL of reaction stop solution was added to each well to terminate the reaction, and the blank well was adjusted to zero. The experimental results were determined within 15 min. The absorbance (A) value at a wavelength of 450 nm was measured by an enzyme-linked detector. Three replicate wells were set for each group of experiments, and the average value was calculated. According to the concentration of the standard and the corresponding absorbance, the linear regression equation of the standard curve was calculated, and then the corresponding sample concentration was calculated on the regression equation according to the absorbance value of the sample. The results are shown in Table 2.

[0060] Table 2 Results of plasma inflammatory factors in rats in each group (pg / mL, n=8)

[0061] Note: Compared with the blank group, a P <0.01; compared with the model group, b P <0.01; compared with drug group 1, c P <0.05.

[0062] As shown in Table 2, the levels of IL-1β, IL-6, IL-8 and TNF-α in the peripheral blood of the model group were significantly increased compared with those of the blank group, with extremely significant differences ( P <0.01), indicating that the model was successfully established. The levels of peripheral blood inflammatory factors IL-1β, IL-6, IL-8 and TNF-α in drug group 1 and drug group 2 were significantly lower than those in the model group, with extremely significant differences ( P <0.01). Drug group 2 was more effective than drug group 1 in reducing inflammatory factors, with significant differences ( P <0.05).

[0063] After the blood was drawn from the abdominal aorta, the rats were quickly dissected and the fat tissue was removed. The weights were weighed with an electronic balance, and the weights of the left and right uteri were weighed and recorded. The uterus was removed and observed with the naked eye. The results are shown in Table 3. The degree of inflammation and swelling of the rat uterine tissue is shown in Table 4. The swelling degree of the rat uterus (g) = left uterus weight - right uterus weight.

[0064] Table 3 Visual inspection results of uterine tissue

[0065] Table 4 Uterine swelling measurement results (g, n=8)

[0066] Note: Compared with the blank group, a P <0.01; compared with the model group, b P <0.01; compared with drug group 1, c P <0.05.

[0067] From the data in the table above, we can see that there is no significant difference in the weight of the left uterus compared with the right uterus in the blank group ( P >0.05). There was a significant difference in the degree of swelling of the left uterus between the model group and the blank group ( P <0.01), indicating that the pelvic inflammatory disease model was successfully established. After administration, the weight and swelling of the left uterus in drug group 1 and drug group 2 were significantly reduced compared with the model group, with extremely significant differences ( P <0.01), and there was a significant difference between drug group 2 and drug group 1 (P<0.05). In summary, the Jindan Fuyan granules produced by drug groups 1 and 2 have different degrees of therapeutic effects on uterine swelling caused by chronic pelvic inflammatory disease, and the Jindan Fuyan granules in drug group 2 have more significant advantages in efficacy than the Jindan Fuyan granules in drug group 1.

[0068] Finally, it should be emphasized that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various changes and modifications. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing Jindan Fuyan granules, characterized in that: The steps include: step 1: extracting Corydalis yanhusuo with vinegar and concentrating the extract into a thick paste for later use; Step 2: extract paeonol from peony bark by steam distillation, and then encapsulate it with β-cyclodextrin to obtain paeonol inclusion complex; collect the water after distillation in another device; Step 3: extract volatile oil from Curcuma zedoaria, Angelica sinensis, Myrrha acetophylla, Cinnamomum cassiae and Cyperus rotundus by steam distillation, then include it with β-cyclodextrin to obtain volatile oil inclusion compound, add gelled silicon dioxide to the volatile oil inclusion compound and mix well, then continue to add hydroxypropyl cellulose and mix well to obtain volatile oil inclusion body; collect the water liquid after distillation in another device; Step 4: Mix the residues after extraction in steps 2 and 3 with honeysuckle, red peony root, Patrinia chinensis, Sargentodoxae, peach kernel, and coix seed, add water and boil, combine the filtrate after filtering with the water solution after distillation in steps 2 and 3, concentrate into a clear paste, precipitate with alcohol, concentrate and combine with the thick paste obtained in step 1, add sucrose and dextrin, and finally add the paeonol inclusion compound obtained in step 2 and the volatile oil inclusion body obtained in step 3, mix well, and granulate to obtain the product.

2. The method for preparing the Jindan Fuyan granules according to claim 1, characterized in that: The raw materials of the Jindan Fuyan granules are as follows by weight: 260 parts of peony bark, 260 parts of honeysuckle, 220 parts of red peony root, 160 parts of curcuma, 200 parts of vinegar Corydalis, 200 parts of Patrinia, 200 parts of Sargentodoxa, 160 parts of peach kernel, 160 parts of angelica, 160 parts of vinegar myrrh, 160 parts of cassia twig, 160 parts of Cyperus rotundus, and 260 parts of coix seed.

3. The method for preparing Jindan Fuyan granules according to claim 1, characterized in that: The colloidal silica in step 3 is SYLOID ® XDP、SYLOID ® 244P or SYLOID ® At least one of 63FP.

4. The method for preparing Jindan Fuyan granules according to claim 1, characterized in that: In step 3, the mass ratio of the volatile oil inclusion compound to the colloidal silicon dioxide is (5-20):

1.

5. The method for preparing Jindan Fuyan granules according to claim 1, characterized in that: In step 3, the viscosity of the hydroxypropyl cellulose is 50-4000 mPa·s, and the molar substitution degree is 3.5-4.

5.

6. The method for preparing Jindan Fuyan granules according to claim 1, characterized in that: In step 3, the hydroxypropyl cellulose needs to be prepared into a solution before being added, and the concentration of the solution is 3%-10%. The solvent used is ethanol, water or a mixture of the two.

7. The method for preparing Jindan Fuyan granules according to claim 6, characterized in that: The weight-to-volume ratio of the mixture obtained by uniformly mixing the volatile oil inclusion compound and the gelled silicon dioxide to the hydroxypropyl cellulose solution is 2:

1.

8. The method for preparing Jindan Fuyan granules according to claim 1, characterized in that: Step 4 also includes packaging the granulated product and filling it with nitrogen.

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