Application of traditional Chinese medicine composition in preparation of medicine for treating depression

Abstract

The invention relates to an application of a traditional Chinese medicine composition in preparation of a medicament for treating depression, which is characterized in that the traditional Chinese medicine composition is prepared from ginseng, rehmannia, cistanche, salvia miltiorrhiza, polygala tenuifolia, rhizoma acori graminei and curcuma aromatica. The composition can obviously improve the depression-like behavior of a depression model mouse, improve the level of nutritional factors in the brain of the depression model mouse and reduce the level of pro-inflammatory cytokines of the depression model mouse, and has a good treatment effect on depression.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology and relates to a new use of a traditional Chinese medicine composition, specifically the application of a traditional Chinese medicine composition in the preparation of a drug for depression. Background Technology

[0002] Depression is a common and serious mental illness characterized by persistent low mood, loss of interest or pleasure, and significant alterations in cognitive, behavioral, and physiological symptoms. Its etiology is complex, involving the interaction of multiple factors, including genetic, biological, psychological, and social environmental factors. Research indicates that dysfunction of neurotransmitters (such as serotonin, norepinephrine, and dopamine) and overactivation of the hypothalamic-pituitary-adrenal (HPA) axis play a crucial role in the development and progression of the disease in patients with depression. Furthermore, psychosocial factors such as chronic stress, traumatic events, and insufficient social support are also considered important triggers for depression.

[0003] The diagnosis of depression is primarily based on the assessment of clinical symptoms. Currently, the internationally widely used diagnostic criteria for mental illness include the Diagnostic and Statistical Manual of Mental Disorders (DSM) and the International Classification of Diseases (ICD). Treatment methods for depression mainly include medication, psychotherapy, and physical therapy. Antidepressants improve symptoms by regulating neurotransmitter levels in the brain, while psychotherapy focuses on helping patients identify and change negative thought patterns and behaviors. In recent years, physical therapy methods such as transcranial magnetic stimulation (TMS) and vagus nerve stimulation (VNS) have also been increasingly applied clinically, providing new treatment options for patients with depression. However, due to the heterogeneity and complexity of depression, the development of individualized treatment plans and long-term management remain challenges in clinical practice. Summary of the Invention

[0004] The first objective of this invention is to provide a traditional Chinese medicine composition for treating depression; the second objective of this invention is to provide a method for preparing the traditional Chinese medicine composition; and the third objective of this invention is to provide the application of the traditional Chinese medicine composition.

[0005] According to a first aspect of the present invention, a traditional Chinese medicine composition for treating depression is provided, wherein its main active ingredients, by weight, include: 1-100 parts ginseng, 1-100 parts rehmannia root, 1-100 parts cistanche, 1-100 parts salvia miltiorrhiza, 1-100 parts polygala tenuifolia, 1-100 parts acorus tatarinowii, and 1-100 parts turmeric. The composition of the present invention can be directly ground into powder, or it can be an extract obtained by conventional means in the art. The traditional Chinese medicines used in the composition of the present invention can also be used by directly grinding into powder, extracting, or other processed forms.

[0006] The preferred weight ratio is: ginseng 9-36 parts, rehmannia glutinosa 20-80 parts, cistanche deserticola 20-80 parts, salvia miltiorrhiza 9-36 parts, polygala tenuifolia 9-36 parts, acorus tatarinowii 9-36 parts, and turmeric 9-36 parts.

[0007] Optionally, the traditional Chinese medicine composition comprises the following components: 9-10 parts ginseng, 18-20 parts rehmannia root, 18-20 parts cistanche, 9-10 parts salvia miltiorrhiza, 9-10 parts polygala tenuifolia, 9-10 parts acorus tatarinowii, and 9-10 parts turmeric.

[0008] In some embodiments, the main active ingredients, by weight, include: 9 parts ginseng, 20 parts rehmannia root, 20 parts cistanche, 9 parts salvia miltiorrhiza, 9 parts polygala tenuifolia, 9 parts acorus tatarinowii, and 9 parts turmeric. In some embodiments, pharmaceutically acceptable excipients are also included. Specifically, the excipients used are carriers or excipients commonly used in the art, including but not limited to starch, lactose, glucose, sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, malt, gelatin, polyols (such as propylene glycol, glycerol, mannitol), tablets, etc., to help improve the stability or activity of the drug, or to produce an acceptable taste or odor when taken orally.

[0009] In some embodiments, the traditional Chinese medicine composition can be prepared as granules, decoctions, tablets, capsules, oral liquids, or pills, etc., thereby facilitating clinical administration.

[0010] According to a second aspect of the present invention, a method for preparing the above-mentioned traditional Chinese medicine composition for depression is provided, the method comprising the following steps: Mix all raw medicinal materials according to the specified ratio, add 8 times the amount of water and soak for 1 hour, extract the volatile oil once for 5 hours, collect the volatile oil by ethyl acetate extraction, and store the ethyl acetate solution and extract of volatile oil separately for later use; recover the ethyl acetate solution of volatile oil at low temperature to obtain volatile oil, store separately for later use; grind and encapsulate the obtained volatile oil with 10 times the amount of β-cyclodextrin and 20 times the amount of water, dry and pulverize to obtain volatile oil inclusion complex for later use; decoct the remaining five ingredients twice with water, 1 hour each time, filter, combine the filtrates, combine the filtrate with the decoction after volatile oil extraction, concentrate to a clear paste with a relative density of 1.08~1.10 (70±5℃), add ethanol to reach an alcohol content of 70%, refrigerate and stand for more than 12 hours, filter, recover the ethanol, and concentrate to a thick paste with a relative density of 1.25~1.30 (70±5℃).

[0011] Furthermore, when the traditional Chinese medicine composition is in the form of granules, dextrin, steviol glycosides, and inclusion complex powder can be added to the above composition for fluidized bed granulation, coating, and drying to obtain the final product.

[0012] According to a third aspect of the present invention, the use of the above-described traditional Chinese medicine composition for depression in the preparation of a medicament for treating depression is provided, wherein "for" can be any means beneficial to improving the corresponding symptoms of the patient, including treatment or prevention.

[0013] Specifically, the traditional Chinese medicine composition, by weight, consists of the following components: 1-100 parts ginseng, 1-100 parts rehmannia root, 1-100 parts cistanche, 1-100 parts salvia miltiorrhiza, 1-100 parts polygala tenuifolia, 1-100 parts acorus tatarinowii, and 1-100 parts turmeric.

[0014] The preferred weight ratio is: ginseng 9-36 parts, rehmannia glutinosa 20-80 parts, cistanche deserticola 20-80 parts, salvia miltiorrhiza 9-36 parts, polygala tenuifolia 9-36 parts, acorus tatarinowii 9-36 parts, and turmeric 9-36 parts.

[0015] Optionally, the traditional Chinese medicine composition comprises the following components: 9-10 parts ginseng, 18-20 parts rehmannia root, 18-20 parts cistanche, 9-10 parts salvia miltiorrhiza, 9-10 parts polygala tenuifolia, 9-10 parts acorus tatarinowii, and 9-10 parts turmeric.

[0016] In some embodiments, the ingredients include: 9 parts ginseng, 20 parts rehmannia root, 20 parts cistanche, 9 parts salvia miltiorrhiza, 9 parts polygala tenuifolia, 9 parts acorus tatarinowii, and 9 parts turmeric.

[0017] Furthermore, when the drug is selected from granules, its preparation method includes the following steps: Mix all raw medicinal materials according to the specified ratio, add 8 times the amount of water and soak for 1 hour, extract the volatile oil once for 5 hours, collect the volatile oil by ethyl acetate extraction, and store the ethyl acetate solution and extract separately for later use; recover the volatile oil from the ethyl acetate solution at low temperature to obtain the volatile oil, and store it separately for later use; grind and encapsulate the obtained volatile oil with 10 times the amount of β-cyclodextrin and 20 times the amount of water, dry and pulverize to obtain the volatile oil inclusion complex for later use; add water to the remaining five ingredients. The decoction is prepared twice, for 1 hour each time. After filtration, the filtrates are combined and then combined with the decoction after extracting the volatile oil. The mixture is concentrated into a clear paste with a relative density of 1.08~1.10 (70±5℃). Ethanol is added until the alcohol content reaches 70%. The paste is then refrigerated and allowed to stand for more than 12 hours. After filtration, the ethanol is recovered and the paste is concentrated into a thick paste with a relative density of 1.25~1.30 (70±5℃). Dextrin, steviol glycosides, and inclusion complex powder are added and the mixture is granulated in a fluidized bed, coated, and dried to obtain the final product.

[0018] The composition of this invention can significantly increase central area movement in open field mice, shorten the immobility time of tail suspension in depressed mice, improve novelty exploration behavior in the elevated cruciate maze, increase BDNF levels in the brain, and decrease the levels of pro-inflammatory cytokines IL-1β and IL-6 in depressed mice. This suggests that the traditional Chinese medicine composition can significantly improve depressive-like mood, increase brain tissue nutritional factors, and inhibit the release of pro-inflammatory factors, thus exhibiting a good therapeutic effect on depression. Detailed Implementation

[0019] As mentioned above, the present invention aims to provide an application of a traditional Chinese medicine composition in the preparation of a drug for depression. The following will describe this in detail with reference to specific experiments.

[0020] Unless otherwise specified, all experiments in the following experiments were conducted under standard conditions or conditions recommended by the manufacturer. The raw materials or excipients used, as well as the reagents or instruments whose manufacturers are not specified, are all commercially available products. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight. Unless otherwise specified, the medicinal materials of this invention can be either raw or processed products, provided that the overall effect of the prescription is not significantly affected.

[0021] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. Example 1: Preparation of Traditional Chinese Medicine Composition

[0022] The traditional Chinese medicine granule composition is made from the following raw materials in parts by weight: ginseng 300g, rehmannia glutinosa 666g, cistanche deserticola 666g, salvia miltiorrhiza 300g, polygala tenuifolia 300g, acorus tatarinowii 300g, and turmeric 300g.

[0023] Mix the above-mentioned raw medicinal materials according to the specified ratio, add 8 times the amount of water and soak for 1 hour. Extract the volatile oil once for 5 hours. Collect the volatile oil by ethyl acetate extraction. Store the ethyl acetate solution and extract of the volatile oil separately for later use. Recover the ethyl acetate solution of the volatile oil at low temperature to obtain the volatile oil, which is then stored separately for later use. Grind and encapsulate the obtained volatile oil with 10 times the amount of β-cyclodextrin and 20 times the amount of water, dry and pulverize to obtain the volatile oil inclusion complex for later use. Decoction the remaining five ingredients with water twice. Each time, for 1 hour, the mixture is filtered, and the filtrates are combined. The filtrates are then combined with the extract of volatile oil and concentrated into a clear paste with a relative density of 1.08~1.10 (70±5℃). Ethanol is added until the alcohol content reaches 70%, and the mixture is refrigerated and allowed to stand for more than 12 hours. The mixture is then filtered, the ethanol is recovered, and the mixture is concentrated into a thick paste with a relative density of 1.25~1.30 (70±5℃) to obtain the traditional Chinese medicine composition. Dextrin, steviol glycosides, and inclusion complex powder are added, and the mixture is granulated in a fluidized bed, coated, and dried to obtain the final product. Experimental Example 2: Therapeutic effect of traditional Chinese medicine composition on CUMS-induced depressive-like behavior in mice

[0024] 1. Experimental Materials 1.1 Animals BALB / C mice, SPF grade, 60 mice, weighing 19-21g, half male and half female.

[0025] Rearing environment: Room temperature 20~26 ºC, relative humidity controlled at 40~70%. Males and females are housed separately, 5 individuals / sex / group per cage.

[0026] 1.2 Drugs The traditional Chinese medicine composition prepared in Example 1 is from Jiangsu Kangyuan Pharmaceutical Co., Ltd.; Fluoxetine Hydrochloride Capsules (20mg / capsule, Eli Lilly Suzhou Pharmaceutical Co., Ltd.) are also mentioned.

[0027] 1.3 Instruments The electronic balance (Sartorius Scientific Instruments, model BS224S). The SuperMzaze analysis system, open field box, elevated cross maze, and forced swimming device were all manufactured by Shanghai Xinruan Information Technology Co., Ltd.

[0028] 2. Dosage design The daily dosage of the herbal composition is 85g of raw drug / day. Based on the body surface area calculation method, the equivalent dose for mice is 85g / 60kg * 12.3 = 17.43g of raw drug / kg. Following a 1 / 4:1 / 2:1 dosing regimen, the low dose is 4.36g of raw drug / kg, and the medium dose is 8.71g of raw drug / kg. Administer by gavage at a dose of 0.2mL / 10g, once daily.

[0029] The daily dose of fluoxetine hydrochloride is 20 mg / day. Based on the body surface area reduction method, the equivalent dose for mice is 20 mg / 60 kg * 12.3 = 4.1 mg / kg. Administer by gavage, 0.2 mL / 10 g, once daily.

[0030] 3. Experimental Methods 3.1 Model Construction Chronic unpredictable stress (CUMS) is currently the most common method for creating mouse models of depression. Mice can exhibit significant depressive-like behaviors under a series of chronic unpredictable mild stresses. In this study, 14 stress conditions were randomly combined, with two stressors selected daily for 28 days. The 14 stress conditions were: 24-hour fasting, 24-hour water deprivation, 24-hour 45-degree tilted cage, reversed day / night cycle, 12-hour continuous light exposure, 24-hour damp bedding, 24-hour removal of bedding, 1 hour of noise (100 dB), 1 hour of foot shock (0.8 mA, 5 s / min), 1 minute of tail clamping, 30 minutes of suspension, 1 hour of cage vibration (220 r / min), 2 hours of body restraint, and 6 minutes of swimming in cold water. The specific experimental arrangements are shown in Table 1.

[0031] Table 1. Experimental Procedure for Chronic Unpredictable Stress

[0032] 3.2 Behavioral Testing (1) Open field experiment: The open field test is a classic unconditioned reflex model widely used to study the neuropsychological state of rodents, primarily testing anxiety and autonomous exploration behaviors in new environments. Mice are gently placed in the center of an open field measuring 50 cm × 50 cm × 50 cm, with the bottom divided into 16 small squares (12.5 cm × 12.5 cm) and the center area consisting of 4 smaller squares. A video capture and analysis system automatically records the mice's autonomous activity for 10 minutes. SuperMzaze software is used to automatically acquire the time each mouse spends in the center area of ​​the open field as analytical data.

[0033] (2) Elevated cross maze experiment: An elevated cross maze (approximately 40cm above the ground) typically contains one open arm (30cm long, 6cm wide) and one closed arm (30cm long, 6cm wide, 15cm high). The frequency and duration of an animal entering the open arm are negatively correlated with anxiety. Lower frequency and shorter duration of entry into the open arm indicate stronger depressive symptoms. The procedure is as follows: (A) Place the mouse in the area formed by the intersection of the two open arms and the two closed arms, with the mouse's head directly facing the open arm. (B) Record the time and number of times the mouse stays on the open arm within five minutes. (C) After each mouse's test, clean the elevated cross maze with an alcohol swab to avoid interference from mouse odor and excrement on the behavior of the next mouse.

[0034] (3) Tail suspension test: Mice were attached to an "S"-shaped hook 1 cm from their tails and hung upside down in a test chamber. The test duration was recorded for 6 minutes. The immobility time of the mice was recorded and analyzed using the SuperMzaze high-throughput tail suspension test analysis system. Data analysis used the immobility time during the 2nd to 6th minute interval.

[0035] 4. Statistical Methods The results were statistically analyzed using t-tests.

[0036] 5. Experimental Results Open field test results showed that, compared with the normal control group, the total distance traveled and the time to enter the central region of the model mice were significantly reduced, with statistically significant differences. P <0.01); After treatment with the traditional Chinese medicine composition, compared with the model group, the total distance of movement and the time to enter the central area were significantly increased in the three traditional Chinese medicine dosage groups, with statistical differences ( P <0.01). See Table 2.

[0037] Table 2: Effects of traditional Chinese medicine composition on the total distance traveled and the number of times the model mice entered the central region in the open field experiment.

[0038] Note: Compared with the normal control group, ** P <0.01; compared with the model group ### P <0.0001.

[0039] The results of the elevated test showed that, compared with the normal control group, the model mice had a significantly reduced time and number of times they entered the open arm, with statistically significant differences. P <0.01); After treatment with the traditional Chinese medicine composition, compared with the model control group, the three traditional Chinese medicine dosage groups showed a significant increase in the time and frequency of entering open arm positions, with statistically significant differences ( P <0.01). See Table 3.

[0040] Table 3: Effects of traditional Chinese medicine composition on the time and number of times a model mouse enters the open arm in the elevated test.

[0041] Note: Compared with the normal control group, *** P <0.001; compared with the model group ### P <0.0001.

[0042] The tail suspension test results showed that, compared with the normal control group, the immobility time of the model mice was significantly increased, with a statistically significant difference. P<0.01); After treatment with the traditional Chinese medicine composition, compared with the model group, the immobility time in the three traditional Chinese medicine dosage groups was significantly reduced, with statistically significant differences ( P <0.01). See Table 4.

[0043] Table 4: Effects of Traditional Chinese Medicine Compositions on Immobility Time in Model Mice during Tail Suspension Test

[0044] Note: Compared with the normal control group, *** P <0.001; compared with the model group ### P <0.0001.

[0045] 6. Experimental Conclusions The traditional Chinese medicine composition can significantly improve anxiety-depression-like behavior in model mice, suggesting that the traditional Chinese medicine composition has a certain therapeutic effect on CUMS-induced depression-like behavior in mice. Experimental Example 3: Protective Effect of Traditional Chinese Medicine Composition on Depression Caused by Chronic Unpredictable Stress

[0046] 1. Experimental Materials 1.1 Test Samples After all behavioral tests were completed, mice were euthanized, and the hippocampus was removed. 0.2 mL of phosphate-buffered saline was added to 20 mg of hippocampal tissue, homogenized, and centrifuged at 3000 rpm for 10 minutes. The supernatant was collected. Blood samples from the mice were also centrifuged at 3000 rpm for 10 minutes. All experimental procedures were strictly performed according to the instructions provided by the ELISA kit manufacturer. The supernatant was stored at -80°C and removed only when needed.

[0047] 1.2 Reagents Mouse corticosterone (CORT) enzyme-linked immunosorbent assay kit (Wuhan Huamei Biotechnology Co., Ltd., CSB-E07969m), mouse interleukin-6 (IL-6) enzyme-linked immunosorbent assay kit (Elabscience, E-EL-M0044), mouse interleukin-1β (IL-1β) enzyme-linked immunosorbent assay kit (Elabscience, E-EL-M0037).

[0048] 1.3 Instruments Low-temperature centrifuge (Eppendorf, Germany, 5804R); microplate reader (McMoRan, USA, Flexstation3).

[0049] 3. Experimental Methods Serum CORT, hippocampal homogenate IL-1β, and IL-6 were detected using ELISA. The experimental procedure was as follows: Standard wells, sample wells, and blank wells were prepared. For the standard wells (7 wells), 100 μL of different concentrations of standard and sample were added sequentially. The plate was covered with a membrane and incubated at 37°C for 1 hour. The liquid was then discarded and the plate was dried. 100 μL of detection solution A (prepared immediately before use) was added to each well. The plate was covered with a membrane and incubated at 37°C for 1 hour. The liquid in the wells was discarded. Each well was washed with 350 μL of washing buffer for 2 minutes. The plate was then gently tapped on absorbent paper to remove all liquid from the wells. This washing process was repeated 3 times. After the final wash, the remaining washing buffer was aspirated or poured out. The plate was inverted on absorbent paper to absorb any remaining liquid from the wells. 100 μL of detection solution B (prepared immediately before use) was added to each well. The plate was covered with a membrane and incubated at 37°C for 30 minutes. Discard the liquid in the wells, shake off excess water, and wash the plate 5 times. Add 90 μL of TMB substrate solution to each well, cover the plate with a membrane, and incubate at 37°C in the dark. Add 50 μL of stop solution to each well to stop the reaction; the blue color will immediately turn yellow. The stop solution should be added in the same order as the substrate solution. After ensuring there are no water droplets on the bottom of the plate and no air bubbles in the wells, immediately measure the optical density of each well at 450 nm using a microplate reader.

[0050] 4. Statistical Methods The results were statistically analyzed using t-tests.

[0051] 5. Experimental Results 5.1 Effects of Traditional Chinese Medicine Composition on Serum CORT in Mice with Chronic Unpredictable Stress-Induced Depression Compared with the control group, the serum CORT level in the model group was significantly increased. Compared with the model group, the CORT level in the fluoxetine hydrochloride group and each example group was significantly decreased, and the differences were statistically significant. P <0.01). See Table 5.

[0052] Table 5: Effects of Traditional Chinese Medicine Compositions on Serum CORT in Mice with Chronic Unpredictable Stress-Induced Depression

[0053] Note: Compared with the normal control group, *** P<0.001; compared with the model group, *** P<0.001. ### P<0.0001.

[0054] 5.2 Effects of Traditional Chinese Medicine Composition on Hippocampal IL-1β and IL-6 in a Mouse Model of Chronic Unpredictable Stress-Induced Depression Compared with the control group, the levels of IL-1β and IL-6 in the hippocampus of the model group were significantly increased, with statistically significant differences. P<0.01). Compared with the model group, the levels of IL-1β and IL-6 in the fluoxetine hydrochloride group and each example group were significantly reduced, and the differences were statistically significant. P <0.01). See Table 6.

[0055] Table 6: Effects of Traditional Chinese Medicine Compositions on Hippocampal IL-1β and IL-6 in a Mouse Model of Depression Caused by Chronic Unpredictable Stress

[0056] Note: Compared with the normal control group, *** P <0.001; compared with the model group ### P <0.0001.

[0057] 6. Experimental Conclusions All three doses of the traditional Chinese medicine composition significantly inhibited elevated glucocorticoid levels in model mice and reduced the levels of IL-1β and IL-6 in the hippocampus, suggesting that the traditional Chinese medicine composition may exert its therapeutic effect on CUMS-induced depression in mice through reducing HPA axis hyperfunction and anti-inflammatory pathways.

Claims

1. The application of a traditional Chinese medicine composition in the preparation of a drug for depression, characterized in that, The traditional Chinese medicine composition is composed of the following components: 1-100 parts of ginseng, 1-100 parts of rehmannia, 1-100 parts of cistanche, 1-100 parts of salvia miltiorrhiza, 1-100 parts of polygala, 1-100 parts of alocasia, and 1-100 parts of curcuma zedoary.

2. Use according to claim 1, characterized in that, The traditional Chinese medicine composition is composed of the following components: 9-36 parts of ginseng, 20-80 parts of rehmannia, 20-80 parts of cistanche, 9-36 parts of salvia miltiorrhiza, 9-36 parts of polygala, 9-36 parts of alocasia, and 9-36 parts of curcuma zedoary.

3. Use according to claim 1, characterized in that, The traditional Chinese medicine composition is composed of the following components: 9-10 parts of ginseng, 18-20 parts of rehmannia, 18-20 parts of cistanche, 9-10 parts of salvia miltiorrhiza, 9-10 parts of polygala, 9-10 parts of alocasia, and 9-10 parts of curcuma zedoary.

4. Use according to claim 1, characterized in that, The traditional Chinese medicine composition is composed of the following components: 9 parts of ginseng, 20 parts of rehmannia, 20 parts of cistanche, 9 parts of salvia miltiorrhiza, 9 parts of polygala, 9 parts of alocasia, and 9 parts of curcuma zedoary.

5. Use according to any one of claims 1 to 4, characterized in that, The medicine for depression also comprises a pharmaceutically acceptable excipient.

6. Use according to claim 5, wherein The pharmaceutically acceptable excipient comprises a filler, a disintegrant, a lubricant, a suspending agent, a binder, a sweetener, a flavoring agent, a preservative, and a base.

7. Use according to claim 5, characterized in that, The dosage form of the medicine for depression is granules, decoction, tablets, capsules, oral liquid, or dripping pills.

8. Use according to any one of claims 1 to 4, characterized in that, The preparation method of the traditional Chinese medicine composition comprises the following steps: The raw medicinal materials are mixed according to the ratio, soaked in 8 times the amount of water for 1 hour, and extracted once to obtain volatile oil, with an extraction time of 5 hours. The volatile oil is extracted with ethyl acetate, and the volatile oil ethyl acetate solution and the extraction liquid are stored separately. The volatile oil ethyl acetate solution is recovered at low temperature to obtain volatile oil, which is also stored separately. The obtained volatile oil is ground with 10 times the amount of β-cyclodextrin and 20 times the amount of water, dried, and pulverized to obtain a volatile oil inclusion compound. The remaining five ingredients are decocted twice with water, each time for 1 hour, filtered, and the filtrates are combined. The filtrate and the extraction liquid are combined, concentrated to a clear paste with a relative density of 1.08-1.10 (70±5°C), added with ethanol to a content of 70%, and refrigerated for 12 hours or more. The ethanol is recovered, and the concentrated thick paste has a relative density of 1.25-1.30 (70±5°C).

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