A kind of peony decoction and its preparation method and application

Abstract

The invention provides a peony decoction and a preparation method and application thereof, belonging to the technical field of traditional Chinese medicine preparations. The preparation method comprises the following steps: S1, by weight, mixing 15-17 parts of coptis root, 2-2.5 parts of senna leaf, 13-15 parts of scutellaria baicalensis and 1.7-2 parts of cinnamon bark, crushing to obtain a mixture, decocting twice, combining the two filtrates, and obtaining an extract; S2, by weight, mixing 25-28 parts of white peony root, 6-7 parts of immature bitter orange, 5-6 parts of Cyperus rotundus and 2.6-3.2 parts of cinnamon bark are mixed and crushed to obtain a second mixture, which is decocted twice and the two filtrates are combined to obtain a second extract; S3, by weight, 12-14 parts of Angelica sinensis, 4-5 parts of Radix Glycyrrhizae Preparata and 1-1.2 parts of cinnamon bark are mixed and crushed to obtain a third mixture, which is decocted twice and the two filtrates are combined to obtain a third extract; S4, the first extract, the second extract and the third extract are mixed and concentrated to obtain Shaoyao Decoction. The present invention can effectively increase blood drug concentration and thus enhance drug efficacy in the case of a fixed dose.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a peony decoction and a preparation method and application thereof. Background Art

[0002] Shaoyao Decoction originates from Liu Wansu's Suwen Bingji Qi Yi Bao Ming Ji. In this formula, Scutellaria baicalensis and Coptis chinensis, both bitter and cold, primarily clear damp-heat in the triple energizers, particularly those accumulating heat and dampness in the large intestine. These herbs serve as the main ingredients. Paeonia lactiflora is heavily used to harmonize the vital energy and blood, alleviating pain, and complements the nourishing and activating effects of Angelica sinensis. Costus root and Areca catechu promote qi circulation and relieve stagnation, regulating the overall qi. Together, these four herbs serve as the assistant ingredients, purging qi and dispelling heaviness, promoting the flow of blood and pus, and healing the body. The bitter and cold purgative properties of Rhubarb complement the heat-clearing and damp-drying properties of Scutellaria baicalensis and Coptis chinensis. A small amount of cinnamon bark aids the circulation of blood and nutrient energy, preventing the bitter and cold effects of Scutellaria and Coptis chinensis from damaging the stomach. Licorice root balances all the ingredients. The entire formula effectively clears heat and dampness, harmonizes qi and blood, and resolves dysentery.

[0003] After oral administration, traditional Chinese medicine (TCM) undergoes four stages of biological activity: absorption, distribution, metabolism, and excretion (ADME). During this process, compounds that are absorbed into the bloodstream and reach a certain plasma concentration are typically key to their pharmacological effects. Therefore, at a fixed dose, increasing the plasma concentration of these key substances is crucial for enhancing efficacy. Summary of the Invention

[0004] In order to solve the problems existing in the background technology, the present invention provides a Shaoyao Decoction and a preparation method and application thereof, which can effectively increase the blood drug concentration and thus enhance the efficacy under the condition of a fixed dose.

[0005] In order to achieve the above objectives, in a first aspect, the present invention provides a method for preparing Shaoyao Decoction, comprising the following steps:

[0006] S1. Mix 15-17 parts of coptis root, 2-2.5 parts of senna leaf, 13-15 parts of scutellaria baicalensis, and 1.7-2 parts of cinnamon bark, by weight, and grind to obtain a mixture 1. The mixture is decocted twice, and the two filtrates are combined to obtain an extract 1.

[0007] S2, by weight, 25-28 parts of white peony root, 6-7 parts of immature bitter orange, 5-6 parts of cyperus rotundus and 2.6-3.2 parts of cinnamon bark were mixed and crushed to obtain a mixture II, which was decocted twice and the two filtrates were combined to obtain an extract II;

[0008] S3. Mix 12-14 parts of angelica sinensis, 4-5 parts of roasted licorice root, and 1-1.2 parts of cinnamon bark by weight, grind them to obtain a mixture of three, decoct them twice, and combine the two filtrates to obtain an extract of three;

[0009] S4. Evenly mix the first extract obtained in S1, the second extract obtained in S2, and the third extract obtained in S3 to obtain a mixed extract, and concentrate the mixed extract to obtain the Shaoyao Decoction.

[0010] Furthermore, in S1, 16 parts of coptis root, 2.3 parts of senna leaves, 14 parts of scutellaria baicalensis and 1.8 parts of cinnamon bark were mixed by weight and crushed to obtain a mixture 1, which was decocted twice and the two filtrates were combined to obtain an extract 1.

[0011] Furthermore, in S2, 26 parts of white peony root, 6.5 parts of immature bitter orange, 5.5 parts of cyperus rotundus and 2.8 parts of cinnamon bark were mixed by weight and crushed to obtain a second mixture, which was decocted twice and the two filtrates were combined to obtain an extract second.

[0012] Furthermore, in S3, 13 parts of angelica sinensis, 4.5 parts of roasted licorice root and 1.1 parts of cinnamon bark were mixed by weight and crushed to obtain a mixture three, which was decocted twice and the two filtrates were combined to obtain an extract three.

[0013] Furthermore, in S1, S2 and S3, the specific operations of decoction twice and combining the two filtrates are the same, and the steps are as follows:

[0014] A1. Add 9-11 times the mass of Mixture 1 / Mixture 2 / Mixture 3 into water, soak, boil, and filter to obtain a first filtrate and a first filter residue;

[0015] A2, adding the first filter residue obtained in A1 into 7-9 times the mass of water, boiling, and filtering to obtain a second filtrate and a second filter residue;

[0016] A3. Mix the first filtrate obtained in A1 and the second filtrate obtained in A2 to obtain the extract 1 / extract 2 / extract 3.

[0017] Furthermore, in A1, the soaking time is 30-40 min; and the boiling time in both A1 and A2 is 1-1.2 h.

[0018] Furthermore, in A1 and A2, 8 layers of 200-mesh medical gauze were used for filtration.

[0019] Furthermore, in S4, the specific operation of the concentration treatment is: concentrating the mixed extract by a rotary evaporator until the mass is equal to the total mass of mixture one, mixture two and mixture three, thereby obtaining the Shaoyao Decoction.

[0020] In a second aspect, the present invention provides a Shaoyao Decoction prepared by the above-mentioned preparation method.

[0021] The effects / efficacy and beneficial effects of each drug component in the present invention are as follows:

[0022] Berberine is the alkaloid with the highest content in Coptis chinensis, which has antibacterial activity and is a quaternary ammonium protoberberine alkaloid among isoquinoline derivatives; coptisine is also a protoberberine quaternary ammonium alkaloid and coexists with berberine in Coptis chinensis; bamipine (palmatine) is the main alkaloid component of Coptis chinensis, the same as the above two alkaloids, and is structurally an isoquinoline derivative.

[0023] Senna leaves contain sennosides (anthraquinone glycosides), which can adjust the polarity of the solvent and enhance the dissolution efficiency of alkaloids such as coptisine and berberine.

[0024] Scutellaria baicalensis contains baicalin and wogonin, both of which are its core flavonoid components, which work together to exert antibacterial, anti-inflammatory, antioxidant and other pharmacological effects.

[0025] On the one hand, the volatile oil components of cinnamon (such as cinnamaldehyde) are warm in nature, which can neutralize the bitter and cold properties of coptis chinensis and scutellaria baicalensis, reduce the stimulation of cold medicines on the spleen and stomach, and maintain the hot and cold balance of the decoction system; on the other hand, volatile oils such as cinnamaldehyde may reduce the degradation of active ingredients caused by acidic or alkaline fluctuations by adjusting the pH value of the decoction system, thereby increasing the dissolution amount and stability of berberine and other ingredients.

[0026] White peony root contains core active ingredients such as paeoniflorin, paeoniflorin, and benzoylpeoniflorin, which have synergistic effects in anti-inflammatory and immune regulation.

[0027] Citrus aurantium contains volatile oil components (such as limonene and β-pinene). Its lipophilicity can reduce the surface tension of the solvent and promote the dispersion and dissolution of fat-soluble components (such as paeoniflorin and benzoylpeoniflorin) in the extraction system. The flavonoids in Citrus aurantium (such as naringin and neohesperidin) have surface activity and can further improve the dissolution efficiency of fat-soluble components (such as paeoniflorin and benzoylpeoniflorin) through emulsification or solubilization mechanisms.

[0028] Cyperus rotundus, the cyclic ketone and α-cyperene in its volatile oil have the function of promoting qi and gastrointestinal motility. It can indirectly promote the release of ingredients such as paeoniflorin by increasing the contact area between the solvent and the medicinal material or accelerating the diffusion of ingredients.

[0029] Cinnamon is also rich in polyphenol antioxidants (such as proanthocyanidins), which can scavenge free radicals and reduce the oxidative stress reaction caused by high temperature during the extraction process, thereby indirectly protecting ingredients such as paeoniflorin that are easily oxidized and degraded, thereby increasing the dissolution amount and stability of paeoniflorin.

[0030] Angelica contains ferulic acid, which has anti-thrombotic and coronary blood flow-increasing effects.

[0031] Roasted licorice contains liquiritin and glycyrrhizin.

[0032] Cinnamon can also combine with phenolic acid components such as ferulic acid and liquiritin through hydrogen bonds or hydrophobic interactions in the extraction system to delay oxidation and improve stability.

[0033] The raw material components of the present invention include coptis root, senna leaf, scutellaria baicalensis, white peony root, immature bitter orange, cyperus rotundus, angelica sinensis, roasted liquorice root and cinnamon bark, and the white peony root and senna leaf are decocted and extracted separately to prepare the peony decoction. In the test example, after the peony decoction is applied to rats, the blood drug concentration data of paeoniflorin, paeoniflorin lactone glycosides and benzoylpeoniflorin measured are all the largest, which is most conducive to improving the dissolution amount, stability and blood drug concentration of paeoniflorin, paeoniflorin lactone glycosides and benzoylpeoniflorin. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 , a comparative trend chart of the test data of paeoniflorin blood concentration after the Shaoyao Decoction prepared in Example 1 and Comparative Examples 1 to Comparative Examples 3 in the test examples of the present invention was applied to rats;

[0035] Figure 2 , a comparative trend chart of test data of blood concentration of paeoniflorin after rats were treated with Shaoyao Decoction prepared in Example 1 and Comparative Examples 1 to 3 in the test examples of the present invention;

[0036] Figure 3 , a comparative trend chart of the test data of benzoylpenoside blood concentration after the Shaoyao Decoction prepared in Example 1 and Comparative Examples 1-3 in the test examples of the present invention was applied to rats. DETAILED DESCRIPTION

[0037] The present application is further described in detail below with reference to the embodiments.

[0038] Unless otherwise specified, the raw materials used in the examples and comparative examples of the present application are all commercially available.

[0039] Example 1: A method for preparing Shaoyao Decoction, comprising the following steps:

[0040] S1. Mix 16 parts of coptis chinensis, 2.3 parts of senna leaves, 14 parts of scutellaria baicalensis, and 1.8 parts of cinnamon bark, and grind them to obtain a mixture one. Put the mixture one into 10 times its mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times its mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract one.

[0041] S2. Mix 26 parts of white peony root, 6.5 parts of immature bitter orange, 5.5 parts of cyperus rotundus and 2.8 parts of cinnamon bark by weight, and grind them to obtain a second mixture. Put the second mixture into 10 times the mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times the mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract two.

[0042] S3. Mix 13 parts of angelica sinensis, 4.5 parts of roasted licorice, and 1.1 parts of cinnamon bark by weight, and grind them to obtain a mixture three. Put the mixture three into 10 times its mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times its mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract three.

[0043] S4. Mix the extract 1 obtained from S1, the extract 2 obtained from S2, and the extract 3 obtained from S3 to obtain a mixed extract, and then concentrate the mixed extract using a rotary evaporator until its mass is equal to the total mass of the mixtures 1, 2, and 3, to obtain Shaoyao Decoction.

[0044] Example 2: The difference between this example and Example 1 is that: a method for preparing Shaoyao Decoction comprises the following steps:

[0045] S1. Mix 15 parts of coptis chinensis, 2 parts of senna leaves, 13 parts of scutellaria baicalensis, and 1.7 parts of cinnamon bark by weight, and grind them to obtain a mixture one. Put the mixture one into 9 times its mass in water, soak for 40 minutes, boil for 1.2 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 7 times its mass in water, boil for 1.2 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract one.

[0046] S2. Mix 25 parts of white peony root, 6 parts of immature bitter orange, 5 parts of cyperus rotundus and 2.6 parts of cinnamon bark by weight, and grind them to obtain a second mixture. Put the second mixture into 9 times the mass of water, soak for 40 minutes, boil for 1.2 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 7 times the mass of water, boil for 1.2 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract two.

[0047] S3. Mix 12 parts of angelica sinensis, 4 parts of roasted licorice, and 1 part of cinnamon bark by weight, and crush them to obtain a mixture three. Put the mixture three into 9 times its mass of water, soak for 40 minutes, boil for 1.2 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 7 times its mass of water, boil for 1.2 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract three.

[0048] S4. Mix the extract 1 obtained from S1, the extract 2 obtained from S2, and the extract 3 obtained from S3 to obtain a mixed extract, and then concentrate the mixed extract using a rotary evaporator until its mass is equal to the total mass of the mixtures 1, 2, and 3, to obtain Shaoyao Decoction.

[0049] Example 3: This example differs from Example 1 in that: a method for preparing Shaoyao Decoction comprises the following steps:

[0050] S1. Mix 17 parts of coptis chinensis, 2.5 parts of senna leaves, 15 parts of scutellaria baicalensis, and 2 parts of cinnamon bark by weight, and grind them to obtain a mixture one. Put the mixture one into 11 times its mass in water, soak for 30 minutes, boil for 1 hour, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 9 times its mass in water, boil for 1 hour, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract one.

[0051] S2. Mix 28 parts of white peony root, 7 parts of immature bitter orange, 6 parts of cyperus rotundus and 3.2 parts of cinnamon bark by weight, and grind them to obtain a second mixture. Put the second mixture into 11 times the mass of water, soak for 30 minutes, boil for 1 hour, and filter through 8 layers of 200 mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 9 times the mass of water, boil for 1 hour, and filter through 8 layers of 200 mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract two.

[0052] S3. Mix 14 parts of angelica sinensis, 5 parts of licorice root, and 1.2 parts of cinnamon bark by weight, and grind them to obtain a mixture three. Put the mixture three into 11 times its mass in water, soak for 30 minutes, boil for 1 hour, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 9 times its mass in water, boil for 1 hour, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract three.

[0053] S4. Mix the extract 1 obtained from S1, the extract 2 obtained from S2, and the extract 3 obtained from S3 to obtain a mixed extract, and then concentrate the mixed extract using a rotary evaporator until its mass is equal to the total mass of the mixtures 1, 2, and 3, to obtain Shaoyao Decoction.

[0054] Comparative Example 1: The difference between this comparative example and Example 1 is that: senna leaves are replaced by rhubarb; and the immature bitter orange and cyperus rotundus are replaced by areca catechu and costus root respectively; and all raw materials are mixed together for extraction.

[0055] Specifically, a method for preparing Shaoyao Decoction comprises the following steps:

[0056] S1. Mix 16 parts of coptis chinensis, 2.3 parts of rhubarb, 14 parts of scutellaria baicalensis, 26 parts of white peony root, 6.5 parts of areca catechu, 5.5 parts of costus root, 13 parts of angelica sinensis, 4.5 parts of roasted liquorice and 5.7 parts of cinnamon bark, and grind them to obtain a mixture. Put the mixture into 10 times the mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times the mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract.

[0057] S2. Concentrate the extract obtained in S1 by rotary evaporation until its mass is equal to that of the mixed material, thereby obtaining Shaoyao Decoction.

[0058] Comparative Example 2: This comparative example differs from Example 1 in that: senna leaves are replaced by rhubarb; and immature bitter orange and cyperus rotundus are replaced by areca catechu and costus root, respectively.

[0059] Specifically, a method for preparing Shaoyao Decoction comprises the following steps:

[0060] S1. Mix 16 parts of coptis chinensis, 2.3 parts of rhubarb, 14 parts of scutellaria baicalensis, and 1.8 parts of cinnamon bark by weight, and grind them to obtain a mixture one. Put the mixture one into 10 times the mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times the mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract one.

[0061] S2. Mix 26 parts of white peony root, 6.5 parts of betel nut, 5.5 parts of costus root and 2.8 parts of cinnamon bark by weight, and grind them to obtain a second mixture. Put the second mixture into 10 times the mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times the mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract two.

[0062] S3. Mix 13 parts of angelica sinensis, 4.5 parts of roasted licorice, and 1.1 parts of cinnamon bark by weight, and grind them to obtain a mixture three. Put the mixture three into 10 times its mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times its mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract three.

[0063] S4. Mix the extract 1 obtained from S1, the extract 2 obtained from S2, and the extract 3 obtained from S3 to obtain a mixed extract, and then concentrate the mixed extract using a rotary evaporator until its mass is equal to the total mass of the mixtures 1, 2, and 3, to obtain Shaoyao Decoction.

[0064] Comparative Example 3: This comparative example differs from Example 1 in that all raw materials are mixed and extracted together.

[0065] Specifically, a method for preparing Shaoyao Decoction comprises the following steps:

[0066] S1. Mix 16 parts of coptis chinensis, 2.3 parts of senna leaves, 14 parts of scutellaria baicalensis, 26 parts of white peony root, 6.5 parts of immature bitter orange, 5.5 parts of cyperus rotundus, 13 parts of angelica sinensis, 4.5 parts of pre-roasted liquorice and 5.7 parts of cinnamon bark, and grind them to obtain a mixture. Put the mixture into 10 times the mass of water, soak for 35 minutes, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first filter residue; put the first filter residue into 8 times the mass of water, boil for 1.1 hours, and filter through 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second filter residue; mix the first filtrate and the second filtrate to obtain an extract.

[0067] S2. Concentrate the extract obtained in S1 by rotary evaporation until its mass is equal to that of the mixed material, thereby obtaining Shaoyao Decoction.

[0068] Experimental Example: 1.1. Experimental instruments: LCMS-8050 ultra-high performance liquid chromatography-tandem quadrupole mass spectrometer (equipped with LC-20A binary gradient pump + SPD-20A UV detector + SIL-20A autosampler, LCMS8050 mass spectrometer (ESI source); Shimadzu Corporation, Japan); Milli-Q ultrapure water analyzer (Millipore Corporation, USA); ultrasound, balance, 5424R high-speed centrifuge (Eppendorf Corporation, Germany).

[0069] 1.2. Drugs and reagents: Coptis chinensis, senna leaf, scutellaria baicalensis, white peony root, immature bitter orange, cyperus rotundus, angelica sinensis, roasted liquorice root and cinnamon bark were purchased from the Traditional Chinese Medicine Clinic of Zhejiang Chinese Medical University.

[0070] Coptisine, berberine, palmatine, wogonin, wogonin, paeoniflorin, paeoniflorin, benzoylpeoniflorin, ferulic acid, glycyrrhizin, and liquiritin were used as indicators of components (analytes) for the pharmacokinetic study in rats.

[0071] Coptisine (batch number C16845532, purity 98%), palmatine (batch number C15813132, purity 98%), wogonin (batch number C15920682, purity 98.02%), and liquiritigenin (batch number C14178361, purity 98%) were purchased from Shanghai Macklin Company; berberine (batch number B10113, purity 98%) was purchased from Beijing Psaitong Company; ferulic acid (batch number YY22101201, purity 98%), paeoniflorin ( Batch number O31GB166251, purity 98%), benzoylpaeoniflorin (batch number A111B222731, purity 98%) were purchased from Shanghai Yuanye Company; wogonin (batch number K2228828, purity 98.71%), paeoniflorin (batch number F2411055, purity 99.56%) were purchased from Shanghai Aladdin Company; liquiritin (batch number BWC9046-2016, purity 99.61%) and acetonitrile were purchased from Merck; mass spectrometry-grade formic acid was purchased from Thermo Fisher Scientific.

[0072] 1.3 Experimental Animals: Male Sprague-Dawley rats (7-8 weeks old, 200 ± 20 g), SPF grade, were purchased from Shanghai Slake Co., Ltd., license number SCXK (Shanghai) 2022-0004. Rats were housed in a 12-h light-dark cycle, maintained at a room temperature of 25 ± 1°C and a humidity of 50 ± 10%, and fed a clean-grade standard rat chow diet. They were fasted for 12 hours before the experiment, but water was not withheld.

[0073] 2.1 Administration: The Shaoyao Decoction prepared in Example 1 and Comparative Examples 1-3 was freeze-dried to obtain freeze-dried Shaoyao Decoction powders, which were then stored at -20°C. Each freeze-dried Shaoyao Decoction powder was prepared with sterilized, single-distilled water to prepare a Shaoyao Decoction solution containing 0.5 g of the crude drug / ml. The solution was administered by oral gavage at a dose of 9.612 g / kg.

[0074] 2.2 Sampling: Rats were randomly divided into the Shaoyao Decoction solution groups (10 rats per group) and fasted for 12 hours with or without water. Each group received corresponding oral administration for 7 days. Immediately after the final oral administration, 0.4 mL of blood was collected from the orbital vein and placed in a sodium heparinized EP tube. The supernatant was centrifuged and stored at -80°C for testing.

[0075] 2.3. Test method: The blood drug concentrations in all the above blood samples were tested using methods known in the art and the average value was taken.

[0076] Among them, the sample processing method is as follows: accurately aspirate 100 μL of plasma sample, add 300 μL of acetonitrile to precipitate protein, vortex for 3 minutes, and then centrifuge at 4°C and 12000 rpm for 15 minutes to remove protein, transfer the supernatant to a clean centrifuge tube, evaporate to dryness with nitrogen purge at 37°C, add 100 μL of 50% acetonitrile to the residue for re-dissolution, vortex for 3 minutes, centrifuge at 4°C and 12000 rpm for 15 minutes to remove a small amount of insoluble matter, and take 1 μL of supernatant for LC-MS / MS analysis.

[0077] LC-MS / MS conditions: Liquid phase system conditions: mobile phase A is 0.1% formic acid water, B is acetonitrile, flow rate is 0.2 mL / min; gradient elution program is: 0-3 min, 30%-30% acetonitrile; 3-30 min, 30%-90% acetonitrile; 30-33 min, 90%-90% acetonitrile; 33-35 min, 90%-30% acetonitrile; 35-55 min, 30%-30% acetonitrile.

[0078] The mass spectrometry system conditions were as follows: an electrospray ionization source (ESI) was used in full scan mode with both positive and negative ions; the scan mode was multiple reaction monitoring (MRM); the nebulizing gas flow rate was 3.0 L / min; the heating gas flow rate was 10 L / min; the interface temperature was 300°C; the DL tube temperature was 250°C; the heating block temperature was 400°C; and the drying gas flow rate was 10 L / min.

[0079] The detection ion pairs and related parameter settings of each compound are shown in Table 1.

[0080] Table 1

[0081]

[0082] 2.3 Test Results: Under the selected chromatographic conditions, the retention times of coptisine, berberine, palmatine, wogonin, and baicalin were 3.12 min, 3.78 min, 3.56 min, 3.21 min, 10.04 min, 2.14 min, 2.15 min, 3.68 min, 2.99 min, 3.844 min, and 2.186 min, respectively. Endogenous substances did not interfere with the assay, demonstrating good method specificity.

[0083] The intra-day and inter-day precision (RSD) values of each compound at high, medium and low concentrations were all less than 15%, and the relative recovery (RR) of each compound at high, medium and low concentrations were between 85% and 115%. The method had good precision and accuracy.

[0084] The blood concentration test data of each compound are shown in Table 2.

[0085] Table 2

[0086]

[0087] Result analysis: Analyze Example 1-Example 3 and Comparative Example 1-Comparative Example 3 and combine the data in Table 2 and Figure 1-Figure 3 From the comparison of the test data of berberine, coptisine and palmatine blood concentrations, it can be seen that compared with rhubarb, senna leaves and coptis chinensis are more conducive to improving the dissolution amount, stability and blood concentration of berberine, coptisine and palmatine when decocted and extracted together.

[0088] By comparing Comparative Example 3 with Example 1, it can be seen that when the raw material components are Coptidis Rhizoma, Senna Leaf, Scutellaria Baicalensis, White Peony Root, Immature Citrus Aurantii, Cyperus Rhizoma, Angelica Sinensis, Preserved Licorice Root and Cinnamon Bark, Comparative Example 3 mixes all the raw material components together for decoction and extraction, while Example 1 decocts and extracts the White Peony Root and Senna Leaf separately. As a result, the blood concentrations of paeoniflorin, paeoniflorin and benzoylpeoniflorin measured after the Peony Decoction prepared in Example 1 acts on rats in the test example are significantly greater than those in Comparative Example 3. This shows that compared with decoction and extraction of White Peony Root and Senna mixed together, decoction and extraction of the two separately can significantly improve the dissolution amount, stability and blood concentration of paeoniflorin, paeoniflorin and benzoylpeoniflorin.

[0089] This is because extracting senna leaves together with white peony root can negatively impact the extraction efficiency and stability of white peony root core components (peoniflorin, peoniflorin, and benzoylpeoniflorin) through mechanisms such as solvent polarity interference, competitive dissolution of components, and oxidative / acidic environmental damage. Specifically, anthraquinone glycosides in senna leaves (such as sennoside A / B) can alter solvent polarity through surface activity, leading to decreased solubility of highly polar monoterpene glycosides such as paeoniflorin and peoniflorin in white peony root under non-ideal polarity conditions. This can even cause precipitation due to altered intermolecular forces, resulting in decreased dissolution of paeoniflorin and peoniflorin. Anthraquinones in senna leaves may also compete with white peony root active components (such as paeoniflorin) for binding sites in the solvent or form complexes, reducing the free concentration of paeoniflorin and inhibiting its dissolution efficiency. During the extraction process, anthraquinone glycosides in senna may also trigger an oxidative chain reaction through free radical pathways, exacerbating the structural destruction of heat-sensitive components (such as paeoniflorin and paeoniflorin), resulting in a decrease in the dissolution amount of paeoniflorin and paeoniflorin.

[0090] By comparing Comparative Example 1 and Comparative Example 2, it can be seen that when the raw material components are Coptis chinensis, Rhubarb, Scutellaria baicalensis, White Peony Root, Areca Catechu, Costus Root, Angelica Sinensis, Roasted Licorice Root and Cinnamon Bark, Comparative Example 1 mixes all the raw material components together for decoction and extraction, while Comparative Example 2 decocts and extracts the White Peony Root and Rhubarb separately. As a result, the blood concentration data of paeoniflorin, paeoniflorin and benzoylpeoniflorin measured after the Peony Decoction prepared in Comparative Example 1 and Comparative Example 2 acted on rats in the test example were very similar, indicating that whether white peony and rhubarb are mixed together for decoction and extraction has no obvious effect on the dissolution amount, stability and blood concentration of paeoniflorin, paeoniflorin and benzoylpeoniflorin.

[0091] This is because the anthraquinone derivatives in rhubarb (such as rhein and emodin) are mostly free anthraquinones, while the content of bound anthraquinones (such as sennosides) is relatively low. Free anthraquinones have low polarity and are less able to modulate solvent polarity than the highly polar anthraquinone glycosides found in senna leaves. Therefore, they have minimal solvent interference with monoterpenoid glycosides in white peony (such as paeoniflorin), resulting in virtually no negative impact. Furthermore, free anthraquinones are relatively weakly hydrophobic, and their effect on solvent polarity is limited, preventing significant inhibition of the solubility of polar components in white peony, such as paeoniflorin.

[0092] As can be seen from the above, senna leaves cannot be decocted and extracted with white peony root, as this would negatively impact the extraction efficiency. Therefore, senna leaves and white peony root should be decocted and extracted separately. Compared to areca nut and costus root, decocting and extracting tangerine peel and cyperus rotundus together with white peony root is more beneficial for improving the dissolution, stability, and blood concentration of paeoniflorin, paeoniflorin, and benzoylpeoniflorin.

[0093] In summary, the raw material components are coptis chinensis, senna leaves, scutellaria baicalensis, white peony root, immature bitter orange, cyperus rotundus, angelica sinensis, roasted licorice root and cinnamon bark, and the white peony root and senna leaves are decocted and extracted separately. As a result, the prepared peony decoction has the highest blood concentration data of paeoniflorin, paeoniflorin and benzoylpeoniflorin after being applied to rats in the experimental example, indicating that it is most conducive to improving the dissolution amount, stability and blood concentration of paeoniflorin, paeoniflorin and benzoylpeoniflorin.

[0094] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0095] In addition, the various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. A method for preparing Shaoyao Decoction, characterized in that: The steps include: S1. Mix 15-17 parts of coptis root, 2-2.5 parts of senna leaf, 13-15 parts of scutellaria baicalensis, and 1.7-2 parts of cinnamon bark, by weight, and grind to obtain a mixture 1. The mixture is decocted twice, and the two filtrates are combined to obtain an extract 1. S2, by weight, 25-28 parts of white peony root, 6-7 parts of immature bitter orange, 5-6 parts of cyperus rotundus and 2.6-3.2 parts of cinnamon bark were mixed and crushed to obtain a mixture II, which was decocted twice and the two filtrates were combined to obtain an extract II; S3. Mix 12-14 parts of angelica sinensis, 4-5 parts of roasted licorice root, and 1-1.2 parts of cinnamon bark by weight, grind them to obtain a mixture of three, decoct them twice, and combine the two filtrates to obtain an extract of three; S4, mixing the first extract obtained in S1, the second extract obtained in S2, and the third extract obtained in S3 to obtain a mixed extract, and concentrating the mixed extract to obtain the Shaoyao Decoction; In S1, S2 and S3, the specific operation of decoction twice and combining the two filtrates is the same, and the steps are as follows: A1. Add 9-11 times the mass of Mixture 1 / Mixture 2 / Mixture 3 into water, soak, boil, and filter to obtain a first filtrate and a first filter residue; A2, adding the first filter residue obtained in A1 into 7-9 times the mass of water, boiling, and filtering to obtain a second filtrate and a second filter residue; A3, mixing the first filtrate obtained in A1 and the second filtrate obtained in A2 to obtain the extract 1 / extract 2 / extract 3; In S4, the specific operation of the concentration treatment is: concentrating the mixed extract by a rotary evaporator until the mass is equal to the total mass of the mixture one, the mixture two and the mixture three, so as to obtain the Shaoyao Decoction.

2. The preparation method of Shaoyao Decoction according to claim 1, characterized in that: In S1, 16 parts of coptis root, 2.3 parts of senna leaves, 14 parts of scutellaria baicalensis and 1.8 parts of cinnamon bark were mixed by weight and crushed to obtain a mixture 1, which was decocted twice and the two filtrates were combined to obtain an extract 1.

3. The preparation method of Shaoyao Decoction according to claim 1, characterized in that: In S2, 26 parts of white peony root, 6.5 parts of immature bitter orange, 5.5 parts of cyperus rotundus and 2.8 parts of cinnamon bark were mixed by weight and crushed to obtain mixed material 2, which was decocted twice and the two filtrates were combined to obtain extract 2.

4. The preparation method of Shaoyao Decoction according to claim 1, characterized in that: In S3, 13 parts of angelica sinensis, 4.5 parts of roasted licorice root and 1.1 parts of cinnamon bark were mixed by weight and crushed to obtain mixture three, which was decocted twice and the two filtrates were combined to obtain extract three.

5. The preparation method of Shaoyao Decoction according to claim 1, characterized in that: In A1, the soaking time was 30-40 min; the boiling time in both A1 and A2 was 1-1.2 h.

6. The preparation method of Shaoyao Decoction according to claim 1, characterized in that: In A1 and A2, 8 layers of 200-mesh medical gauze were used for filtration.

7. A peony decoction, characterized in that: The method is as described in any one of claims 1 to 6.

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