Paeonia lactiflora soup as well as preparation method and application thereof

Abstract

The invention provides Chinese herbaceous peony soup and a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine preparations, the preparation method comprises the following steps: S1, mixing 15-17 parts by weight of coptis chinensis, 2-2.5 parts by weight of folium sennae, 13-15 parts by weight of scutellaria baicalensis and 1.7-2 parts by weight of cinnamon, crushing to obtain a mixture I, decocting twice, and combining the filtrates of the two times to obtain an extracting solution I; s2, 25-28 parts by weight of radix paeoniae alba, 6-7 parts by weight of fructus aurantii immaturus, 5-6 parts by weight of rhizoma cyperi and 2.6-3.2 parts by weight of cortex cinnamomi are mixed and smashed, a second mixture is obtained and decocted twice, filtrate obtained after two times of decoction is combined, and a second extracting solution is obtained; s3, 12-14 parts by weight of angelica sinensis, 4-5 parts by weight of honey-fried licorice roots and 1-1.2 parts by weight of cinnamon are mixed and smashed, a third mixture is obtained and decocted twice, filtrate obtained after two times of decoction is combined, and a third extracting solution is obtained; s4, uniformly mixing the extracting solution I, the extracting solution II and the extracting solution III, and concentrating to obtain the Chinese herbaceous peony soup. Under the condition of fixed dosage, the blood concentration can be effectively improved, and the drug effect is further enhanced.

Description

Technical Field

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

[0002] Shaoyao Decoction comes from Liu Wansu's Suwen Bingji Qi Yi Baoming Ji. The two herbs in the prescription, Scutellaria baicalensis and Coptis chinensis, are bitter and cold in nature. They are mainly used to clear away the damp-heat poison in the three burners and are good at treating the heat and dampness poison in the large intestine. They are the main herbs. Paeonia lactiflora is used to harmonize the blood and relieve pain. It is combined with Angelica sinensis to nourish blood and activate blood circulation. Aucklandia lappa and Areca nut can promote qi and relieve stagnation, regulate the qi of the whole body, and the four herbs are used together to achieve the meaning of "regulating qi and removing heaviness in the back, and promoting blood and pus to heal by itself". They are all ministerial herbs. The bitter and cold purgative effect of rhubarb is used to assist Scutellaria baicalensis and Coptis chinensis in clearing heat and drying dampness. A small amount of cinnamon is used together to help Angelica dahurica and Peony Root to promote blood circulation and harmonize the blood and prevent the bitter and cold Scutellaria baicalensis and Coptis chinensis from damaging the stomach. They are all adjuvant herbs. Licorice harmonizes all the herbs. The whole prescription plays the role of clearing heat and drying dampness, regulating qi and blood, and curing dysentery.

[0003] After Chinese medicine is taken orally and enters the body, the process of exerting its biological effect goes through four stages: absorption, distribution, metabolism and excretion (ADME). In this process, compounds that can be absorbed into the blood and reach a certain blood concentration are usually the key substances to exert pharmacological effects. Therefore, under the condition of a fixed dose, how to increase the blood concentration of these key substances is the key to enhancing the 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-mentioned object, in a first aspect, the present invention provides a method for preparing Shaoyao Decoction, comprising the following steps: 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, grind to obtain a mixture 1, decoct twice, combine the two filtrates, and 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, by weight, 12-14 parts of angelica, 4-5 parts of licorice and 1-1.2 parts of cinnamon are mixed and crushed to obtain a mixture three, and the mixture is decocted twice and the two filtrates are combined to obtain an extract 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.

[0006] Further, in S1, 16 parts by weight of Coptis chinensis, 2.3 parts by weight of Senna leaf, 14 parts by weight of Scutellaria baicalensis, and 1.8 parts by weight of Cinnamon are mixed and pulverized to obtain a first mixture. After two decoctions and combining the two filtrates, a first extract is obtained.

[0007] Further, in S2, 26 parts by weight of Paeonia lactiflora, 6.5 parts by weight of Fructus Aurantii Immaturus, 5.5 parts by weight of Cyperus rotundus, and 2.8 parts by weight of Cinnamon are mixed and pulverized to obtain a second mixture. After two decoctions and combining the two filtrates, a second extract is obtained.

[0008] Further, in S3, 13 parts by weight of Angelica sinensis, 4.5 parts by weight of Radix Glycyrrhizae Preparata, and 1.1 parts by weight of Cinnamon are mixed and pulverized to obtain a third mixture. After two decoctions and combining the two filtrates, a third extract is obtained.

[0009] Further, in S1, S2, and S3, the specific operations of the two decoctions and combining the two filtrates are the same, and the steps are as follows: A1. Put the first mixture / second mixture / third mixture into water with a mass 9 - 11 times that of the mixture, soak, decoct, and filter to obtain a first filtrate and a first residue; A2. Put the first residue obtained in A1 into water with a mass 7 - 9 times that of the residue, decoct, and filter to obtain a second filtrate and a second residue; A3. Mix the first filtrate obtained in A1 and the second filtrate obtained in A2 to obtain the first extract / second extract / third extract.

[0010] Further, in A1, the soaking time is 30 - 40 min; the decocting time in both A1 and A2 is 1 - 1.2 h.

[0011] Further, in A1 and A2, 8 - layer medical gauze with 200 meshes is used for filtration.

[0012] Further, in S4, the specific operation of the concentration treatment is: Concentrate the mixed extract to the same mass as the total mass of the first mixture, second mixture, and third mixture through a rotary evaporator to obtain the Paeonia Decoction.

[0013] In the second aspect, the present invention provides a Paeonia Decoction prepared by the above - mentioned preparation method.

[0014] The functions / effects and beneficial effects of each drug component in the present invention are as follows: Coptis chinensis, Berberine is the alkaloid with the highest content, has antibacterial activity, and belongs to the quaternary ammonium type proto - berberine type alkaloid in the isoquinoline derivative category; Coptisine also belongs to the proto - berberine type quaternary alkaloid and co - exists with berberine in Coptis chinensis; Palmatine is also one of the main alkaloid components of Coptis chinensis together with the above two alkaloids and belongs to the isoquinoline derivative in terms of structure.

[0015] Senna leaf, containing sennosides (anthraquinone glycosides), can adjust the polarity of the solvent and enhance the dissolution efficiency of alkaloids such as coptisine and berberine.

[0016] Scutellaria baicalensis contains baicalin and wogonin, both of which are its core flavonoid components and jointly participate in exerting pharmacological effects such as antibacterial, anti-inflammatory, and antioxidant effects.

[0017] Cinnamon, on the one hand, its volatile oil components (such as cinnamaldehyde) are warm in nature, can neutralize the bitter-cold properties of Coptis chinensis and Scutellaria baicalensis, reduce the stimulation of cold-natured drugs to the spleen and stomach, and at the same time maintain the cold-heat 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, etc.

[0018] White peony root contains core active ingredients such as albiflorin, paeoniflorin, and benzoylpaeoniflorin, and these ingredients have synergistic effects in aspects such as anti-inflammatory and immunomodulation.

[0019] Immature bitter orange contains volatile oil components (such as limonene, β-pinene), and its lipophilicity can reduce the surface tension of the solvent and promote the dispersion and dissolution of lipophilic components (such as albiflorin, benzoylpaeoniflorin) in the extraction system; the flavonoid substances (such as naringin, neohesperidin) in immature bitter orange have surface activity effects and can further increase the dissolution efficiency of lipophilic components (such as albiflorin, benzoylpaeoniflorin) through emulsification or solubilization mechanisms. Cyperus rotundus, the cyperone and α-cyperene in its volatile oil have the effects of promoting qi movement and gastrointestinal motility, and can indirectly promote the release of components such as albiflorin by increasing the contact area between the solvent and the medicinal materials or accelerating the diffusion of components. Cinnamon is also rich in polyphenolic antioxidant substances (such as procyanidins), which can scavenge free radicals, reduce the oxidative stress reaction caused by high temperature during the extraction process, thereby indirectly protecting easily oxidized and degraded components such as paeoniflorin, and further increasing the dissolution amount and stability of paeoniflorin, etc.

[0020] Angelica sinensis contains ferulic acid, which has the effects of anti-thrombosis and increasing coronary blood flow. Prepared licorice root contains liquiritin and liquiritigenin.

[0021] Cinnamon can also combine with phenolic acid components such as ferulic acid and liquiritin through hydrogen bonding or hydrophobic interaction in the extraction system, delay oxidation, and improve stability.

[0022] The raw material components of the present invention are Coptis chinensis, Senna leaf, Scutellaria baicalensis, Paeonia lactiflora, Immature Bitter Orange, Cyperus rotundus, Angelica sinensis, Honey-fried Licorice Root, and Cinnamon. The Paeonia lactiflora and Senna leaf are decocted and extracted separately. The paeoniflorin, albiflorin, and benzoylpaeoniflorin blood drug concentration data measured after the prepared Shaoyao Decoction acts on rats in the test examples are the largest, which is most conducive to improving the dissolution, stability, and blood drug concentration of paeoniflorin, albiflorin, and benzoylpaeoniflorin. Description of the Drawings

[0023] Figure 1 、Comparison trend chart of paeoniflorin blood drug concentration test data after the Shaoyao Decoction prepared in Example 1 and Comparative Examples 1 - 3 of the present invention acts on rats; Figure 2 、Comparison trend chart of albiflorin blood drug concentration test data after the Shaoyao Decoction prepared in Example 1 and Comparative Examples 1 - 3 of the present invention acts on rats; Figure 3 、Comparison trend chart of benzoylpaeoniflorin blood drug concentration test data after the Shaoyao Decoction prepared in Example 1 and Comparative Examples 1 - 3 of the present invention acts on rats. Detailed Description of the Invention

[0024] The following further elaborates on the present application in conjunction with the examples.

[0025] The raw materials of the examples and comparative examples of the present application are all commonly commercially available unless otherwise specified.

[0026] Example 1: A preparation method of Shaoyao Decoction, comprising the following steps: S1. By weight, mix 16 parts of Coptis chinensis, 2.3 parts of Senna leaf, 14 parts of Scutellaria baicalensis, and 1.8 parts of Cinnamon, pulverize to obtain mixture one. Put mixture one into water with a mass 10 times that of the mixture, soak for 35 min, decoct for 1.1 h, and then filter with 8 - layer 200 - mesh medical gauze to obtain the first filtrate and the first residue; put the first residue into water with a mass 8 times that of the residue, decoct for 1.1 h, and then filter with 8 - layer 200 - mesh medical gauze to obtain the second filtrate and the second residue; mix the first filtrate and the second filtrate to obtain extract one.

[0027] S2. By weight, mix 26 parts of Paeonia lactiflora, 6.5 parts of Immature Bitter Orange, 5.5 parts of Cyperus rotundus, and 2.8 parts of Cinnamon, pulverize to obtain mixture two. Put mixture two into water with a mass 10 times that of the mixture, soak for 35 min, decoct for 1.1 h, and then filter with 8 - layer 200 - mesh medical gauze to obtain the first filtrate and the first residue; put the first residue into water with a mass 8 times that of the residue, decoct for 1.1 h, and then filter with 8 - layer 200 - mesh medical gauze to obtain the second filtrate and the second residue; mix the first filtrate and the second filtrate to obtain extract two.

[0028] S3. Mix 13 parts by weight of Angelica sinensis, 4.5 parts of roasted licorice, and 1.1 parts of cinnamon, crush them to obtain mixture three. Put mixture three into water with a mass 10 times that of mixture three, soak for 35 minutes, boil for 1.1 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 8 times that of the first filter residue, boil for 1.1 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract three.

[0029] S4. Mix extract one obtained in S1, extract two obtained in S2, and extract three obtained in S3 to obtain a mixed extract. Then, concentrate the mixed extract by a rotary evaporator until its mass is equal to the total mass of mixture one, mixture two, and mixture three, and thus the Paeonia Decoction is obtained.

[0030] Example 2: The difference between this example and Example 1 lies in: A preparation method of Paeonia Decoction, including the following steps: S1. Mix 15 parts by weight of Coptis chinensis, 2 parts of Senna leaf, 13 parts of Scutellaria baicalensis, and 1.7 parts of cinnamon, crush them to obtain mixture one. Put mixture one into water with a mass 9 times that of mixture one, soak for 40 minutes, boil for 1.2 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 7 times that of the first filter residue, boil for 1.2 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract one.

[0031] S2. Mix 25 parts by weight of Paeonia lactiflora, 6 parts of Fructus Aurantii Immaturus, 5 parts of Cyperus rotundus, and 2.6 parts of cinnamon, crush them to obtain mixture two. Put mixture two into water with a mass 9 times that of mixture two, soak for 40 minutes, boil for 1.2 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 7 times that of the first filter residue, boil for 1.2 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract two.

[0032] S3. Mix 12 parts by weight of Angelica sinensis, 4 parts of roasted licorice, and 1 part of cinnamon, crush them to obtain mixture three. Put mixture three into water with a mass 9 times that of mixture three, soak for 40 minutes, boil for 1.2 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 7 times that of the first filter residue, boil for 1.2 hours, and then filter through 8 layers of 200-mesh medical gauze to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract three.

[0033] S4. Mix the extract 1 obtained in S1, the extract 2 obtained in S2, and the extract 3 obtained in S3 to obtain a mixed extract, and then concentrate the mixed extract by a rotary evaporator to a mass equal to the total mass of the mixture 1, the mixture 2, and the mixture 3, thereby obtaining the Shaoyao Decoction.

[0034] Example 3: The difference between this example and Example 1 is as follows: A method for preparing the Shaoyao Decoction, comprising the following steps: S1. By weight, mix 17 parts of Coptis chinensis, 2.5 parts of Senna leaf, 15 parts of Scutellaria baicalensis, and 2 parts of Cinnamon, and pulverize to obtain mixture 1. Put mixture 1 into water with a mass 11 times that of mixture 1, soak for 30 min, boil for 1 h, and then filter using 8 layers of 200-mesh medical gauze to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 9 times that of the first filter residue, boil for 1 h, and then filter using 8 layers of 200-mesh medical gauze to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract 1.

[0035] S2. By weight, mix 28 parts of White Peony Root, 7 parts of Immature Bitter Orange, 6 parts of Cyperus rotundus, and 3.2 parts of Cinnamon, and pulverize to obtain mixture 2. Put mixture 2 into water with a mass 11 times that of mixture 2, soak for 30 min, boil for 1 h, and then filter using 8 layers of 200-mesh medical gauze to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 9 times that of the first filter residue, boil for 1 h, and then filter using 8 layers of 200-mesh medical gauze to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract 2.

[0036] S3. By weight, mix 14 parts of Chinese Angelica, 5 parts of Prepared Licorice Root, and 1.2 parts of Cinnamon, and pulverize to obtain mixture 3. Put mixture 3 into water with a mass 11 times that of mixture 3, soak for 30 min, boil for 1 h, and then filter using 8 layers of 200-mesh medical gauze to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 9 times that of the first filter residue, boil for 1 h, and then filter using 8 layers of 200-mesh medical gauze to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract 3.

[0037] S4. Mix the extract 1 obtained in S1, the extract 2 obtained in S2, and the extract 3 obtained in S3 to obtain a mixed extract, and then concentrate the mixed extract by a rotary evaporator to a mass equal to the total mass of the mixture 1, the mixture 2, and the mixture 3, thereby obtaining the Shaoyao Decoction.

[0038] Comparative Example 1: The difference between this comparative example and Example 1 is as follows: Senna leaf is replaced by Rheum officinale; and Immature Bitter Orange and Cyperus rotundus are respectively replaced by Areca catechu and Aucklandia lappa; and all raw materials are mixed for extraction together.

[0039] Specifically, a method for preparing the Shaoyao Decoction, comprising the following steps: S1. By weight, mix 16 parts of Coptis chinensis, 2.3 parts of Rheum palmatum, 14 parts of Scutellaria baicalensis, 26 parts of Paeonia lactiflora, 6.5 parts of Areca catechu, 5.5 parts of Aucklandia lappa, 13 parts of Angelica sinensis, 4.5 parts of prepared licorice root, and 5.7 parts of Cinnamomum cassia, pulverize to obtain a mixed material. Put the mixed material into water with a mass 10 times that of the mixed material, soak for 35 min, decoct for 1.1 h, and then filter with 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first residue; put the first residue into water with a mass 8 times that of the first residue, decoct for 1.1 h, and then filter with 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second residue; mix the first filtrate and the second filtrate to obtain an extract.

[0040] S2. Concentrate the extract obtained in S1 to the same mass as the mixed material by a rotary evaporator to obtain Shaoyao Decoction.

[0041] Comparative Example 2: The difference between this comparative example and Example 1 is that Folium Sennae is replaced by Rheum palmatum; and Aurantii Fructus Immaturus and Cyperus rotundus are replaced by Areca catechu and Aucklandia lappa, respectively.

[0042] Specifically, a preparation method of Shaoyao Decoction includes the following steps: S1. By weight, mix 16 parts of Coptis chinensis, 2.3 parts of Rheum palmatum, 14 parts of Scutellaria baicalensis, and 1.8 parts of Cinnamomum cassia, pulverize to obtain a first mixed material. Put the first mixed material into water with a mass 10 times that of the first mixed material, soak for 35 min, decoct for 1.1 h, and then filter with 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first residue; put the first residue into water with a mass 8 times that of the first residue, decoct for 1.1 h, and then filter with 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second residue; mix the first filtrate and the second filtrate to obtain a first extract.

[0043] S2. By weight, mix 26 parts of Paeonia lactiflora, 6.5 parts of Areca catechu, 5.5 parts of Aucklandia lappa, and 2.8 parts of Cinnamomum cassia, pulverize to obtain a second mixed material. Put the second mixed material into water with a mass 10 times that of the second mixed material, soak for 35 min, decoct for 1.1 h, and then filter with 8 layers of 200-mesh medical gauze to obtain a first filtrate and a first residue; put the first residue into water with a mass 8 times that of the first residue, decoct for 1.1 h, and then filter with 8 layers of 200-mesh medical gauze to obtain a second filtrate and a second residue; mix the first filtrate and the second filtrate to obtain a second extract.

[0044] S3. Mix 13 parts by weight of Angelica sinensis, 4.5 parts of roasted licorice, and 1.1 parts of cinnamon, pulverize them to obtain mixture three. Put mixture three into water with a mass 10 times that of mixture three, soak for 35 min, decoct for 1.1 h, and then filter with 8-layer medical gauze of 200 meshes to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 8 times that of the first filter residue, decoct for 1.1 h, and then filter with 8-layer medical gauze of 200 meshes to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain extract three.

[0045] S4. Mix extract one obtained in S1, extract two obtained in S2, and extract three obtained in S3 to obtain a mixed extract, and then concentrate the mixed extract by a rotary evaporator to a mass equal to the total mass of mixture one, mixture two, and mixture three, thus obtaining the Paeony Decoction.

[0046] Comparative Example 3: The difference between this comparative example and Example 1 is that all raw materials are mixed for extraction together.

[0047] Specifically, a preparation method of Paeony Decoction includes the following steps: S1. Mix 16 parts by weight of Coptis chinensis, 2.3 parts of Senna leaf, 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 roasted licorice, and 5.7 parts of cinnamon, pulverize them to obtain a mixture. Put the mixture into water with a mass 10 times that of the mixture, soak for 35 min, decoct for 1.1 h, and then filter with 8-layer medical gauze of 200 meshes to obtain the first filtrate and the first filter residue. Put the first filter residue into water with a mass 8 times that of the first filter residue, decoct for 1.1 h, and then filter with 8-layer medical gauze of 200 meshes to obtain the second filtrate and the second filter residue. Mix the first filtrate and the second filtrate to obtain an extract.

[0048] S2. Concentrate the extract obtained in S1 by a rotary evaporator to a mass equal to that of the mixture, thus obtaining the Paeony Decoction.

[0049] Test 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 ultraviolet detector + SIL-20A automatic sampler, LCMS8050 mass spectrometer (ESI source); Shimadzu Corporation, Japan); Milli-Q ultrapure water instrument (Millipore Corporation, USA); ultrasonic device, balance, 5424R high-speed centrifuge (Eppendorf AG, Germany).

[0050] 1.2. Drugs and reagents: Coptis chinensis, Senna leaf, Scutellaria baicalensis, white peony root, immature bitter orange, Cyperus rotundus, Angelica sinensis, roasted licorice, and cinnamon are all purchased from the Traditional Chinese Medicine Clinic of Zhejiang Chinese Medical University.

[0051] Coptisine, berberine, palmatine, wogonoside, wogonin, albiflorin, paeoniflorin, benzoylpaeoniflorin, ferulic acid, liquiritigenin, and liquiritin were used as the indices of the components (analytes) for the pharmacokinetic study in rats.

[0052] Coptisine (batch number C16845532, purity 98%), palmatine (batch number C15813132, purity 98%), wogonoside (batch number C15920682, purity 98.02%), and liquiritigenin (batch number C14178361, purity 98%) were purchased from Shanghai Macklin Biochemical Co., Ltd.; berberine (batch number B10113, purity 98%) was purchased from Beijing psaitong Biotechnology Co., Ltd.; ferulic acid (batch number YY22101201, purity 98%), albiflorin (batch number O31GB166251, purity 98%), and benzoylpaeoniflorin (batch number A11lB222731, purity 98%) were purchased from Shanghai Yuanye Bio-Technology Co., Ltd.; wogonin (batch number K2228828, purity 98.71%) and paeoniflorin (batch number F2411055, purity 99.56%) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; liquiritin (batch number BWC9046-2016, purity 99.61%) and acetonitrile were purchased from Merck KGaA; formic acid of mass spectrometry grade was purchased from Thermo Fisher Scientific.

[0053] 1.3 Experimental animals: SD rats (male, 7 - 8 weeks old, 200 ± 20 g), SPF grade, were purchased from Shanghai SLAC Laboratory Animal Co., Ltd., with the license number SCXK (Hu) 2022 - 0004. The rats were housed under a 12 h light / dark cycle and maintained at an indoor temperature of 25 ± 1°C and a humidity of 50 ± 10%. They were fed with clean-grade ordinary rat feed. The rats were fasted for 12 h before the experiment, but water was not restricted.

[0054] 2.1 Administration: The Shaoyao Decoctions prepared in Example 1 and Comparative Examples 1 - 3 were freeze-dried respectively to obtain freeze-dried powders of Shaoyao Decoction, which were stored in a -20°C refrigerator. Each freeze-dried powder of Shaoyao Decoction was dissolved in sterilized distilled water to prepare a Shaoyao Decoction solution containing 0.5 g of crude drug per milliliter. The drug was administered by gavage at a dose of 9.612 g / kg.

[0055] 2.2 Sampling: The rats were randomly divided into groups of the Shaoyao Decoction solutions of Example 1 and Comparative Examples 1 - 3 (10 rats in each group), and they were fasted for 12 h without water restriction; each group was given the drug by gavage for 7 d. After the last gavage, 0.4 mL of blood was immediately collected from the orbital vein of the rats and placed in heparinized EP tubes. The supernatant was obtained by centrifugation and stored in a -80°C refrigerator for further testing.

[0056] 2.3 Testing method: The blood drug concentrations in all the above blood samples were tested by the methods known in the art, and the average values were taken.

[0057] Among them, the sample treatment method: precisely pipette 100 μL of plasma sample, add 300 μL of acetonitrile to precipitate proteins, vortex for 3 min, then centrifuge at 4°C and 12,000 rpm for 15 min to remove proteins. Transfer the supernatant to a clean centrifuge tube, evaporate to dryness under nitrogen blowing at 37°C. Dissolve the residue in 100 μL of 50% acetonitrile, vortex for 3 min, centrifuge at 4°C and 12,000 rpm for 15 min to remove a small amount of insoluble substances, and take 1 μL of the supernatant for LC-MS / MS analysis.

[0058] LC-MS / MS conditions: Liquid phase system conditions: Mobile phase A is 0.1% formic acid in water, B is acetonitrile, and the flow rate is 0.2 mL / min; The gradient elution program is as follows: 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.

[0059] Mass spectrometry system conditions: Electrospray ionization source (ESI) is used, with positive and negative ion full scan mode; The scanning mode is multiple reaction monitoring (MRM); The nebulizing gas flow rate is 3.0 L / min; The heating gas flow rate is 10 L / min; The interface temperature is 300°C; The DL tube temperature is 250°C; The heating block temperature is 400°C; The drying gas flow rate is 10 L / min.

[0060] The detection ion pairs and related parameter settings of each compound are shown in Table 1.

[0061] Table 1

[0062] 2.3. Test results: Under the selected chromatographic conditions, the retention time of coptisine is 3.12 min, the retention time of berberine is 3.78 min, the retention time of palmatine is 3.56 min, the retention time of wogonoside is 3.21 min, the retention time of wogonin is 10.04 min, the retention time of albiflorin is 2.14 min, the retention time of paeoniflorin is 2.15 min, the retention time of benzoylpaeoniflorin is 3.68 min, the retention time of ferulic acid is 2.99 min, the retention time of liquiritigenin is 3.844 min, and the retention time of liquiritin is 2.186 min. Endogenous substances do not interfere with the determined components, and the method has good specificity.

[0063] For each compound at high, medium, and low concentrations, the within-day and between-day precision (RSD) values are all less than 15%, and the relative recovery rates (RR) of each compound at high, medium, and low concentrations are all between 85% - 115%. The method has good precision and accuracy.

[0064] The test data of the blood drug concentration of each compound are shown in Table 2.

[0065] Table 2

[0066] Result analysis: Analyze Examples 1 - 3 and Comparative Examples 1 - 3 and combine the data in Table 2 and Figures 1 - 3 , from the comparison of the test data of the blood drug concentration of berberine, coptisine and palmatine, it can be seen that compared with rhubarb, when senna leaf is decocted and extracted together with Coptis chinensis, it is more beneficial to improve the dissolution amount, stability and blood drug concentration of berberine, coptisine and palmatine.

[0067] From the comparison between Comparative Example 3 and Example 1, when the raw material components are Coptis chinensis, senna leaf, Scutellaria baicalensis, Paeonia lactiflora, Fructus Aurantii Immaturus, Cyperus rotundus, Angelica sinensis, Radix Glycyrrhizae Preparata and Cinnamomum cassia, in Comparative Example 3, all the raw material components are mixed and decocted together, while in Example 1, Paeonia lactiflora and senna leaf are decocted separately. As a result, the blood drug concentration data of paeoniflorin, albiflorin and benzoylpaeoniflorin measured after the Paeonia decoction prepared in Example 1 acts on rats in the test example are significantly higher than those in Comparative Example 3. It shows that compared with decocting and extracting Paeonia lactiflora and senna leaf together, decocting them separately can significantly improve the dissolution amount, stability and blood drug concentration of paeoniflorin, albiflorin and benzoylpaeoniflorin.

[0068] This is because when senna leaf is extracted together with Paeonia lactiflora, it will have a negative impact on the extraction efficiency and stability of the core components of Paeonia lactiflora (paeoniflorin, albiflorin, benzoylpaeoniflorin) through mechanisms such as solvent polarity interference, component competitive dissolution, and oxidation / acidic environment destruction. Specifically, the anthraquinone glycoside components (such as sennoside A / B) in senna leaf can change the solvent polarity through surface activity, resulting in a decrease in the solubility of the monoterpene glycoside components with higher polarity such as paeoniflorin and albiflorin in Paeonia lactiflora in a non-ideal polarity environment, and even precipitation due to changes in intermolecular forces, resulting in a decrease in the dissolution amount of paeoniflorin, albiflorin, etc. The anthraquinone substances in senna leaf and the active components of Paeonia lactiflora (such as paeoniflorin) may also compete for the binding sites in the solvent or form complexes, reducing the free concentration of paeoniflorin and inhibiting its dissolution efficiency. The anthraquinone glycoside components in senna leaf may also trigger an oxidative chain reaction through a free radical pathway during the extraction process, exacerbating the structural damage of heat-sensitive components (such as paeoniflorin and albiflorin), resulting in a decrease in the dissolution amount of paeoniflorin, albiflorin, etc.

[0069] Comparing Comparative Example 1 and Comparative Example 2, when the raw material components are Coptis chinensis, Rheum palmatum, Scutellaria baicalensis, Paeonia lactiflora, Areca catechu, Aucklandia lappa, Angelica sinensis, Radix Glycyrrhizae Preparata, and Cinnamomum cassia, in Comparative Example 1, all the raw material components were mixed and decocted together, while in Comparative Example 2, Paeonia lactiflora and Rheum palmatum were decocted separately. As a result, the blood drug concentration data of paeoniflorin, albiflorin, and benzoylpaeoniflorin measured after the paeonia decoction prepared in Comparative Example 1 and Comparative Example 2 acted on rats in the test examples were very similar, indicating that whether Paeonia lactiflora and Rheum palmatum are mixed and decocted together has no obvious effect on the dissolution amount, stability, and blood drug concentration of paeoniflorin, albiflorin, and benzoylpaeoniflorin.

[0070] This is because most of the anthraquinone derivatives (such as rhein and emodin) in Rheum palmatum are free anthraquinones, while the content of combined anthraquinones (such as sennosides) is relatively low. Free anthraquinones have a relatively low polarity and a weaker ability to regulate the polarity of the solvent than the high-polarity anthraquinone glycosides in Senna leaves. Therefore, the interference of the solvent on the monoterpene glycosides (such as paeoniflorin) in Paeonia lactiflora is very small and basically has no negative impact. Moreover, the hydrophobicity of free anthraquinones is relatively weak, and the change range of the solvent polarity is very limited, so the solubility of polar components such as paeoniflorin in Paeonia lactiflora will not be significantly inhibited.

[0071] As can be seen from the above, Senna leaves cannot be decocted together with Paeonia lactiflora, otherwise it will have a negative impact on the extraction effect of Paeonia lactiflora. Therefore, Senna leaves and Paeonia lactiflora need to be decocted separately. Compared with Areca catechu and Aucklandia lappa, when Fructus Aurantii Immaturus and Cyperus rotundus are decocted together with Paeonia lactiflora, it is more beneficial to improve the dissolution amount, stability, and blood drug concentration of paeoniflorin, albiflorin, and benzoylpaeoniflorin.

[0072] In summary, when the raw material components are Coptis chinensis, Senna leaves, Scutellaria baicalensis, Paeonia lactiflora, Fructus Aurantii Immaturus, Cyperus rotundus, Angelica sinensis, Radix Glycyrrhizae Preparata, and Cinnamomum cassia, and Paeonia lactiflora and Senna leaves are decocted separately, the blood drug concentration data of paeoniflorin, albiflorin, and benzoylpaeoniflorin measured after the paeonia decoction prepared acts on rats in the test examples are all the largest, indicating that it is most beneficial to improve the dissolution amount, stability, and blood drug concentration of paeoniflorin, albiflorin, and benzoylpaeoniflorin.

[0073] In addition, it should be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without conflict. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

[0074] In addition, any combination can be made between the various different embodiments of the present invention, as long as it does not violate the idea of the present invention, it should also be regarded as the content disclosed by the present invention.

Claims

1. A preparation method of Shaoyao Decoction, characterized in that, It includes the following steps: S1. By weight parts, mix 15 - 17 parts of Coptis chinensis, 2 - 2.5 parts of Senna leaf, 13 - 15 parts of Scutellaria baicalensis, and 1.7 - 2 parts of Cinnamon, pulverize to obtain the first mixture, decoct twice, combine the two filtrates to obtain the first extract; S2. By weight parts, mix 25 - 28 parts of Paeonia lactiflora, 6 - 7 parts of Fructus Aurantii Immaturus, 5 - 6 parts of Cyperus rotundus, and 2.6 - 3.2 parts of Cinnamon, pulverize to obtain the second mixture, decoct twice, combine the two filtrates to obtain the second extract; S3. By weight parts, mix 12 - 14 parts of Angelica sinensis, 4 - 5 parts of Radix Glycyrrhizae Preparata, and 1 - 1.2 parts of Cinnamon, pulverize to obtain the third mixture, decoct twice, combine the two filtrates to obtain the third extract; S4. Mix the first extract obtained in S1, the second extract obtained in S2, and the third extract obtained in S3, obtain the mixed extract, and perform concentration treatment to obtain the said Shaoyao Decoction.

2. The preparation method of the Paeonia Decoction according to claim 1, wherein, In S1, by weight parts, mix 16 parts of Coptis chinensis, 2.3 parts of Senna leaf, 14 parts of Scutellaria baicalensis, and 1.8 parts of Cinnamon, pulverize to obtain the first mixture, decoct twice, combine the two filtrates to obtain the first extract.

3. The preparation method of Shaoyao Decoction according to claim 1, characterized in that In S2, by weight parts, mix 26 parts of Paeonia lactiflora, 6.5 parts of Fructus Aurantii Immaturus, 5.5 parts of Cyperus rotundus, and 2.8 parts of Cinnamon, pulverize to obtain the second mixture, decoct twice, combine the two filtrates to obtain the second extract.

4. The preparation method of the Paeonia Decoction according to claim 1, wherein, In S3, by weight parts, mix 13 parts of Angelica sinensis, 4.5 parts of Radix Glycyrrhizae Preparata, and 1.1 parts of Cinnamon, pulverize to obtain the third mixture, decoct twice, combine the two filtrates to obtain the third extract.

5. The preparation method of the Paeonia Decoction according to claim 1, wherein, In S1, S2, and S3, the specific operations of decocting twice and combining the two filtrates are the same, and the steps are as follows: A1. Put the first mixture / the second mixture / the third mixture into water with a mass 9 - 11 times that of the mixture, soak, decoct, and filter to obtain the first filtrate and the first residue; A2. Put the first residue obtained in A1 into water with a mass 7 - 9 times that of the residue, decoct, and filter to obtain the second filtrate and the second residue; A3. Mix the first filtrate obtained in A1 and the second filtrate obtained in A2 to obtain the said first extract / second extract / third extract.

6. The preparation method of the Paeonia Decoction according to claim 5, wherein, In A1, the soaking time is 30 - 40 min; the decocting times in A1 and A2 are both 1 - 1.2 h.

7. The preparation method of the Paeonia decoction according to claim 5, characterized in that, In A1 and A2, filter with 8 - layer medical gauze of 200 meshes.

8. The preparation method of the Paeonia Decoction according to claim 1, wherein In S4, the specific operation of the concentration treatment is: Concentrate the mixed extract by a rotary evaporator to a mass equal to the total mass of the first mixture, the second mixture, and the third mixture to obtain the said Shaoyao Decoction.

9. A Paeonia decoction, characterized in that, Prepared by the preparation method described in any one of claims 1 - 8.

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