Traditional Chinese medicine composition for treating depression caused by stagnation of liver qi and phlegm obstructing heart orifices, and preparation method and quality control method of traditional Chinese medicine composition for treating depression caused by stagnation of liver qi and phlegm obstructing heart orifices

Abstract

The invention provides a traditional Chinese medicine composition for treating depression caused by stagnation of liver qi and phlegm obstructing heart orifices, and a preparation method and a quality control method thereof, and belongs to the technical field of traditional Chinese medicines. On one hand, the invention provides a traditional Chinese medicine composition formula for treating depression caused by stagnation of liver qi and stagnation of phlegm in heart orifices; secondly, the invention provides a traditional Chinese medicine composition for treating depression caused by stagnation of liver qi and stagnation of phlegm in heart orifices; thirdly, the invention provides a preparation method of the traditional Chinese medicine composition for treating depression caused by stagnation of liver qi and phlegm obstructing heart orifices. Fourthly, the invention further provides application of the traditional Chinese medicine composition in preparation of a medicine for treating depression. In the fifth aspect, the invention further provides a fingerprint spectrum construction method of the traditional Chinese medicine composition for treating depression caused by stagnation of liver qi and phlegm obstructing heart orifices. In the sixth aspect, the invention also provides application of the fingerprint spectrum in drug detection and quality control for treating depression.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a traditional Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices, a preparation method and a quality control method thereof. Background Art

[0002] Depression is a mental disorder characterized by low mood and loss of pleasure, accompanied by cognitive, behavioral or autonomic nervous system symptoms, which significantly affects individual functions. It is a mental illness that seriously threatens human physical and mental health, with high morbidity, high disability rate, high suicide rate and high recurrence rate. It is estimated that by 2030, depression will surpass tumors and cardiovascular and cerebrovascular diseases to become the world's largest disease burden.

[0003] Currently, there are limited treatment options for depressive disorders. About 30% to 40% of patients become refractory to depression due to insensitivity to existing monoamine antidepressants. Monoamine antidepressants also have shortcomings such as slow onset of action, residual symptoms, more adverse reactions, and the need for long-term medication. New rapid antidepressants also have problems with drug addiction and neurotoxicity, and treatment faces new difficulties. "Guidelines for the Clinical Application of Traditional Chinese Medicines in the Treatment of Depressive Disorders (2022)"

[0004] The etiology and pathogenesis of depressive disorders are complex and have not yet been fully elucidated. Therefore, modern medicine still faces bottlenecks in the treatment of depressive disorders. At the same time, commonly used antidepressants have delayed efficacy and multiple adverse reactions, which seriously affect medication compliance.

[0005] In clinical diagnosis and treatment practice, TCM syndrome differentiation and treatment has unique advantages and has fewer side effects than Western medicine. At the same time, compared with traditional Chinese medicine decoctions, solid preparations of Chinese medicine have the advantages of being easy to take, carry and store, and are more convenient for patients to accept. For example, Anle Capsule (main ingredients: Polygonum multiflorum, Bupleurum, Angelica, Chuanxiong, Poria, Uncaria, Stir-fried Atractylodes, Licorice) uses Chinese medicine such as Polygonum multiflorum to achieve the effects of soothing the liver, relieving depression, and calming the mind. However, Anle Capsule is more oriented to menopausal depression and has a smaller scope of application.

[0006] The patent application with publication number CN102406917A discloses an antidepressant Chinese medicine composition and its preparation method, including Morinda officinalis, Bupleurum chinense, White Peony Root, Roasted Citrus aurantium, Atractylodes macrocephala (fried), Pinellia ternata, Polygala tenuifolia (processed), Licorice (fried), Arisaema consanguineum, Curcuma aromatica, Ginger, Ziziphus jujuba seed (fried), and Poria cocos.

[0007] Traditional Chinese medicine has always been difficult for patients to accept because of its bitter, astringent, and spicy taste, especially the Polygala root, which "hurts the throat". However, this herb has the effects of calming the mind, improving intelligence, and connecting the heart and kidneys. It is used for insomnia, dreaminess, forgetfulness, palpitations, and confusion, and has a very good effect. It occupies an important position in the prescription. Therefore, it is urgent to find a preparation method to reduce or remove the bad taste of this product while ensuring the efficacy.

[0008] In addition, the rough preparation process of traditional antidepressant Chinese medicine restricts the efficacy of the medicine.

[0009] In view of this, the present invention provides a Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and mind, a preparation method and a quality control method thereof. Summary of the invention

[0010] This prescription contains 2 parts of bupleurum, 3 parts of white peony root, 2 parts of polygala, 2 parts of poria, 2 parts of tangerine peel, 2 parts of julibrissin bark, 2 parts of lily, 1 part of schisandra chinensis, and 1 part of roasted liquorice.

[0011] In the formula, Bupleurum and White Peony are the main drugs. Bupleurum is bitter, pungent and slightly cold, which can dispel fever, relieve liver qi and relieve depression, and raise Yang Qi. White Peony is bitter, sour and slightly cold, which can restrain Yin and stop sweating, soften the liver and relieve pain, and suppress liver Yang. The two drugs are used together, which are interdependent and mutually promoted, and each other's shortcomings are restrained and their strengths are developed. Bupleurum's pungent and dispersing properties can soothe the liver and regulate Qi, and the white peony's sour and astringent properties can be used to guide the drugs directly to the Shaoyang meridian, and nourish blood to moisten its orifices and benefit its meridians. One Qi and one blood regulate Qi and blood stagnation, suppress liver Yang, and clear the gallbladder and soothe the liver, reconcile the exterior and interior, raise Yang and restrain Yin, and relieve depression and relieve pain.

[0012] Polygala, Poria, and Dictyophora are all ministerial drugs. Polygala enters the heart, kidney, and lung meridians, can calm the mind and improve intelligence, and connect the heart and kidney. It is pungent and warm, and enters the heart and spleen meridians, can open the orifices, eliminate filth, and relieve pain. Poria is sweet, light, and flat. It enters the heart, lung, spleen, and kidney meridians, promotes diuresis and eliminates dampness, strengthens the spleen, and calms the mind; Dictyophora is pungent, bitter, and warm, and enters the lung and spleen meridians, regulates qi and relieves fullness, dries dampness and resolves phlegm. The combination of the four ministerial drugs can clear phlegm and refresh the mind, and quickly relieve symptoms such as chest fullness, flank pain, irritability, anxiety, and foreign body sensation in the throat.

[0013] Albizzia julibrissin bark, lily and Schisandra chinensis are adjuvants. Albizzia julibrissin bark enters the heart, liver and lung meridians, and can relieve depression and calm the mind. Lily is sweet and cold, and enters the heart and lung meridians, and can nourish yin and moisten the lungs, clear the heart and calm the mind. Schisandra chinensis enters the lung, heart and kidney meridians, and can calm the mind and calm the mind. The three medicines used together can relieve depression and calm the mind, and assist in the treatment of symptoms such as mental confusion, restlessness, palpitations and insomnia, dizziness and tinnitus.

[0014] Roasted licorice root is a guiding drug, which nourishes the spleen and stomach, relieves pain, and harmonizes various medicines.

[0015] The purpose of the present invention is to provide a Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices, a preparation method and a quality control method thereof.

[0016] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0017] In a first aspect, the present invention provides a Chinese medicine composition formula for treating depression caused by liver qi stagnation and phlegm blocking the heart and mind, the Chinese medicine composition formula is composed of the following components: bupleurum, white peony root, polygala, poria, tangerine peel, julibrissin bark, lily, schisandra chinensis, and roasted liquorice.

[0018] Furthermore, the Chinese medicine composition formula consists of the following components in parts by weight: 2 parts of bupleurum, 3 parts of white peony root, 2 parts of polygala, 2 parts of poria, 2 parts of tangerine peel, 2 parts of julibrissin bark, 2 parts of lily, 1 part of schisandra chinensis, and 1 part of roasted liquorice.

[0019] In a second aspect, the present invention provides a traditional Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and mind. The traditional Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and mind is composed of the following components: water extracts of white peony root, Poria cocos, Citrus aurantium, Albizzia julibrissin bark, Lilium bulbus, and Radix Glycyrrhizae Radix et Rhizoma, alcohol extracts of Bupleurum chinense and Schisandra chinensis, and enzymatic acid water extracts of Bupleurum chinense and Schisandra chinensis residues and Polygala tenuifolia slices.

[0020] Furthermore, the Chinese medicine composition is obtained from the following raw materials in parts by weight: 2 parts of bupleurum, 3 parts of white peony root, 2 parts of polygala, 2 parts of poria, 2 parts of tangerine peel, 2 parts of julibrissin bark, 2 parts of lily, 1 part of schisandra chinensis, and 1 part of roasted liquorice.

[0021] In a third aspect, the present invention provides a method for preparing a Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices, the preparation method comprising the following steps:

[0022] White peony root, Poria cocos, tangerine peel, Albizzia julibrissin, lily, and licorice root are extracted with water, and the liquid is dried to obtain a water extract of white peony root, Poria cocos, tangerine peel, Albizzia julibrissin, lily, and licorice root; Bupleurum root and Schisandra chinensis are extracted by reflux, and the liquid is dried to obtain a Bupleurum root and Schisandra chinensis alcohol extract; Bupleurum root and Schisandra chinensis residues and coarse powder of Polygala tenuifolia are hydrolyzed with water, then boiled and extracted, and the liquid is dried to obtain a hydrolyzed acid water extract of Bupleurum root and Schisandra chinensis residues and Polygala tenuifolia root;

[0023] The above-mentioned white peony root, Poria, Citrus aurantium, Albizzia julibrissin, Lilium and Radix Glycyrrhizae water extracts, Bupleurum and Schisandrae Chinensis alcohol extracts, Bupleurum and Schisandrae Chinensis residues and Polygala tenuifolia decoction pieces are mixed to obtain a composition.

[0024] Furthermore, the specific steps of the preparation method of the Chinese medicine composition are:

[0025] Add 8-10 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 6-8 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0026] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice, and 7 to 9 times the amount of 85 to 95% ethanol was added for the first time, and the extraction was performed for 1 hour, and the extraction was filtered. The second time, 6 to 8 times the amount of 85 to 95% ethanol was added, and the extraction was performed for 1 hour, and the extraction was filtered. The combined medicinal solution was concentrated under reduced pressure at 50 to 70°C and dried, and the dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0027] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae are extracted with water twice, the first time the amount is 8 to 12 times, the pH value is adjusted to 4.3 to 5.3, 1000 to 3000 U of pectinase and 1000 to 2000 U of cellulase are added per kilogram of the decoction pieces, and the enzymatic hydrolysis is carried out at 40 to 50° C. for 5 to 7 hours, and then the extraction is boiled for 1 hour and centrifuged, the second time the amount is 7 to 9 times, the pH value is adjusted to 4.3 to 5.3, the extraction is boiled for 1 hour and centrifuged, the combined medicinal solution is concentrated under reduced pressure at 60 to 90° C. and dried, the dry paste is crushed and passed through a 60-mesh sieve to obtain the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae;

[0028] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, Lilium and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition.

[0029] Furthermore, in a preferred embodiment of the present invention, the preparation method of the Chinese medicine composition comprises the following specific steps:

[0030] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0031] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0032] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of pectinase and 1500U of cellulase were added per kg of the decoction pieces, the enzymatic hydrolysis was carried out at 45°C for 6 hours, and then the extraction was boiled for 1 hour, and centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, the extraction was boiled for 1 hour, and centrifuged, the combined medicinal solution was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0033] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, Lilium and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition.

[0034] In a fourth aspect, the present invention also provides a use of the above composition in the preparation of a drug for treating depression.

[0035] In a fifth aspect, the present invention also provides a method for constructing a fingerprint of a Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices, the fingerprint construction method comprising the following steps:

[0036] Take gallic acid, astragaloside, paeoniflorin, lily A, paeoniflorin, sibirica polygala xanthoside A, polygala xanthoside III, naringin, 3,6'-diesinapoylsucrose, polygala tenuifolia glycoside A, licorice glycoside, benzoylpeoniflorin, ammonium glycyrrhizinate, saikosaponin b1, schisandrae alcohol A reference substances to prepare reference substance solution;

[0037] Take a Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices, and prepare a test solution of the Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices;

[0038] The test solution and the reference solution of the Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices are respectively subjected to high performance liquid chromatography analysis to construct a fingerprint of the Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices;

[0039] The conditions for high performance liquid chromatography analysis include: acetonitrile as mobile phase A and 0.15% phosphoric acid solution as mobile phase B; gradient elution; the gradient elution program includes: 0min to 10min, the volume percentage of mobile phase A changes from 5% to 10%; 10min to 25min, the volume percentage of mobile phase A changes from 10% to 20%; 25min to 30min, the volume percentage of mobile phase A changes from 20% to 25%; 30min to 35min, the volume percentage of mobile phase A changes from 25% to 28%; 35min to 50min, the volume percentage of mobile phase A changes from 28% to 40%; 50min to 65min, the volume percentage of mobile phase A changes from 40% to 55%.

[0040] Furthermore, the fingerprint of the traditional Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and mind comprises 15 common peaks; among them, taking peak 8 naringin as the reference peak, the relative retention time of peak 1 gallic acid is 0.221±10%, the relative retention time of peak 2 astragaloside is 0.519±10%, the relative retention time of peak 3 paeoniflorin is 0.686±10%, the relative retention time of peak 4 royal lily A is 0.705±10%, the relative retention time of peak 5 paeoniflorin is 0.740±10%, the relative retention time of peak 6 Siberian polygala xanthoconone A is 0.814±10%, and the relative retention time of peak 7 polygala is 0.806±10%. The relative retention time of orthoxanthenone III was 0.875±10%, the relative retention time of peak 9 3,6'-diesinapoylsucrose was 1.019±10%, the relative retention time of peak 10 polygalaglycoside A was 1.092±10%, the relative retention time of peak 11 liquiritigenin was 1.128±10%, the relative retention time of peak 12 benzoylpeonidin was 1.364±10%, the relative retention time of peak 13 ammonium glycyrrhizinate was 1.669±10%, the relative retention time of peak 14 saikosaponin b1 was 1.745±10%, and the relative retention time of peak 15 schisandrin A was 1.782±10%.

[0041] In a sixth aspect, the present invention also provides an application of a fingerprint as a standard fingerprint in drug detection and quality control for the treatment of depression.

[0042] Compared with the prior art, the beneficial effects of the present invention include at least:

[0043] 1. The extraction method combining reflux and enzymatic hydrolysis is adopted to extract the effective ingredients and remove the impurities that cause bad smell and taste, so that the prepared Chinese medicine composition preparation has a mild taste and is easy for patients to take.

[0044] 2. Establish a characteristic spectrum method and verify the method. At the same time, combine UPLC-Q-TOF-MS analysis and reference substances to confirm the attribution and identification of characteristic peaks to ensure that the quality control method is stable and reliable, highly specific, and has clear markers, which can be used for quality control of this product. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 Overlay of HPLC chromatograms.

[0046] Figure 2 It is a 3D picture.

[0047] Figure 3 It is an isoabsorption spectrum.

[0048] Figure 4 Comparative HPLC spectra for peak identification. DETAILED DESCRIPTION

[0049] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0050] The features and properties of the present invention are further described in detail below in conjunction with the embodiments:

[0051] 1. Composition and preparation method thereof Examples and comparative examples

[0052] Example 1

[0053] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0054] Add 8 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 6 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0055] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. The first time, 7 times the amount of 95% ethanol was added, extracted for 1 hour, filtered, and the second time, 8 times the amount of 85% ethanol was added, extracted for 1 hour, filtered, and the combined liquid was concentrated under reduced pressure at 50°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0056] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 8 times, the pH value was adjusted to 5.3, 1000U of pectinase and 2000U of cellulase were added per kg of the decoction pieces, the enzymatic hydrolysis was carried out at 40°C for 7 hours, and then the extraction was boiled for 1 hour, centrifuged, the second time the amount was 7 times, the pH value was adjusted to 5.3, the extraction was boiled for 1 hour, centrifuged, the combined medicinal solution was concentrated under reduced pressure at 60°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0057] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0058] Example 2

[0059] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0060] Add 10 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 8 times the amount of water to the residue and decoct for 30 minutes, filter, concentrate and dry the combined liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0061] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 9 times the amount of 85% ethanol was added, extracted for 1 hour, filtered, and for the second time, 6 times the amount of 95% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0062] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 12 times, the pH value was adjusted to 4.3, 3000U of pectinase and 1000U of cellulase were added per kg of the decoction pieces, the enzymatic hydrolysis was carried out at 50°C for 5 hours, and then the extraction was boiled for 1 hour, and the extraction was centrifuged, the second time the amount was 9 times, the pH value was adjusted to 4.3, the extraction was boiled for 1 hour, and the extraction was centrifuged, the combined medicinal solution was concentrated under reduced pressure at 90°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0063] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0064] Example 3

[0065] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0066] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0067] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0068] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of pectinase and 1500U of cellulase were added per kg of the decoction pieces, the enzymatic hydrolysis was carried out at 45°C for 6 hours, and then the extraction was boiled for 1 hour, and centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, the extraction was boiled for 1 hour, and centrifuged, the combined medicinal solution was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0069] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0070] Comparative Example 1

[0071] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0072] Take the above-mentioned pieces, add 9 times the amount of water, boil for 60 minutes, filter, add 7 times the amount of water to the residue, boil for 30 minutes, filter, concentrate and dry the combined medicinal liquid, crush the dry paste and pass it through a 60-mesh sieve to obtain a water extract;

[0073] The above water extract is added with auxiliary material dextrin for granulation, dried, granulated and bagged to obtain granules.

[0074] Comparative Example 2

[0075] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0076] Take the above-mentioned crude powder of the decoction pieces, reflux extract twice, add 8 times the amount of 90% ethanol for the first time, extract for 1 hour, filter, add 7 times the amount of 90% ethanol for the second time, extract for 1 hour, filter, the combined medicinal solution is concentrated under reduced pressure at 60°C and dried, the dry paste is crushed and passed through a 60-mesh sieve to obtain an alcohol extract;

[0077] The above alcohol extract is added with auxiliary material dextrin for granulation, dried, granulated and bagged to obtain granules.

[0078] Comparative Example 3

[0079] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0080] Add 9 times the amount of water to the pieces of Radix Bupleuri, Poria, Huajuhong, Albizzia julibrissin, Lilium and Radix Glycyrrhizae, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of Radix Bupleuri, Poria, Huajuhong, Albizzia julibrissin, Lilium and Radix Glycyrrhizae;

[0081] The crude powder of white peony root and Schisandra chinensis slices were refluxed and extracted twice. The first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined liquid was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the white peony root and Schisandra chinensis alcohol extract;

[0082] The white peony root, Schisandra chinensis dregs and Polygala tenuifolia decoction pieces were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of pectinase and 1500U of cellulase were added per kg of decoction pieces, and the mixture was hydrolyzed at 45°C for 6 hours, then boiled and extracted for 1 hour, centrifuged, and the second time the amount was 8 times, the pH value was adjusted to 4.5, boiled and extracted for 1 hour, centrifuged, the combined liquid was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the white peony root, Schisandra chinensis dregs and Polygala tenuifolia decoction pieces was obtained;

[0083] The above-mentioned water extracts of bupleurum, poria, tangerine peel, julibrissin bark, lily, and licorice, the alcohol extracts of white peony root and schisandra chinensis, and the enzymatic acid water extracts of the residues of white peony root and schisandra chinensis and polygala tenuifolia are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate, and bag to obtain granules.

[0084] Comparative Example 4

[0085] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0086] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0087] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0088] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of pectinase and 1500U of cellulase were added per kg of the decoction pieces, the enzymatic hydrolysis was carried out at 45°C for 6 hours, and then the extraction was boiled for 1 hour, and centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, the extraction was boiled for 1 hour, and centrifuged, the combined medicinal solution was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0089] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0090] Comparative Example 5

[0091] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0092] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0093] The crude powder of Polygala, Bupleurum and Schisandra pieces were refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined liquid was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the alcohol extract of Polygala, Bupleurum and Schisandra;

[0094] Polygala, Bupleurum and Schisandra chinensis residues were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of pectinase and 1500U of cellulase were added per kg of the slices, enzymolysis was performed at 45°C for 6 hours, then boiled and extracted for 1 hour, centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, boiled and extracted for 1 hour, centrifuged, the combined liquid was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of Polygala, Bupleurum and Schisandra chinensis residues was obtained;

[0095] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, Lilium and roasted liquorice, alcohol extracts of Polygala tenuifolia, Bupleurum chinense and Schisandra chinensis, and enzymatic acid water extracts of Polygala tenuifolia, Bupleurum chinense and Schisandra chinensis residues are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0096] Comparative Example 6

[0097] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0098] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0099] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0100] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 1500U of cellulase was added per kg of the decoction pieces, enzymolysis was performed at 45°C for 6 hours, then boiled and extracted for 1 hour, centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, boiled and extracted for 1 hour, centrifuged, the combined medicinal solution was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0101] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0102] Comparative Example 7

[0103] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0104] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0105] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0106] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of pectinase was added per kg of the slices, enzymolysis was performed at 45°C for 6 hours, then boiled and extracted for 1 hour, centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, boiled and extracted for 1 hour, centrifuged, the combined liquid was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the Radix Polygalae was obtained;

[0107] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0108] Comparative Example 8

[0109] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0110] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0111] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0112] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of pectinase and 1500U of ligninase were added per kg of the decoction pieces, the enzymatic hydrolysis was carried out at 45°C for 6 hours, and then the extraction was boiled for 1 hour, and centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, the extraction was boiled for 1 hour, and centrifuged, the combined medicinal solution was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0113] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0114] Comparative Example 9

[0115] Bupleurum 100g, White Peony 150g, Polygala 100g, Poria 100g, Huajuhong 100g, Albizzia Julibrissin 100g, Lilium 100g, Schisandra 50g, Roasted Licorice 50g

[0116] Add 9 times the amount of water to the pieces of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root, decoct for 60 minutes, filter, add 7 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the water extract of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root;

[0117] The crude powder of Bupleurum and Schisandra slices was refluxed and extracted twice. For the first time, 8 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and for the second time, 7 times the amount of 90% ethanol was added, extracted for 1 hour, filtered, and the combined medicinal solution was concentrated under reduced pressure at 60°C and dried. The dry paste was crushed and passed through a 60-mesh sieve to obtain the Bupleurum and Schisandra alcohol extract;

[0118] The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae were extracted with water twice, the first time the amount was 10 times, the pH value was adjusted to 4.8, 2000U of ligninase and 1500U of cellulase were added per kg of the decoction pieces, the enzymatic hydrolysis was carried out at 45°C for 6 hours, and then the extraction was boiled for 1 hour, and centrifuged, the second time the amount was 8 times, the pH value was adjusted to 4.5, the extraction was boiled for 1 hour, and centrifuged, the combined liquid was concentrated under reduced pressure at 75°C and dried, the dry paste was crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae was obtained;

[0119] The above-mentioned water extracts of white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and roasted liquorice, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala tenuifolia slices are mixed to obtain a composition; the auxiliary material dextrin is added to the composition to granulate, dry, granulate and bag to obtain granules.

[0120] 2. Quality Control Method Implementation

[0121] (1) Methodology

[0122] ① Choice of mobile phase

[0123] After screening multiple mobile phase systems, types, and proportions, and through chromatographic separation, asymmetry, peak shape and other indicators, the results determined that the following chromatographic conditions had good chromatographic peak separation and moderate response values, which can be used as detection conditions for the characteristic spectrum of this product.

[0124] Table 1 Elution conditions

[0125]

[0126]

[0127] ② Detection wavelength selection

[0128] The diode array detector was used to scan the test solution at a wavelength of 190-400nm, and the 3D view and isoabsorption spectrum were used for analysis. The results showed that the number of chromatographic peaks was the largest in the wavelength range of 200-250nm. In this wavelength range, the chromatograms at wavelengths of 200nm, 210nm, 220nm, 225nm, 230nm, 235nm, 240nm, 245nm, and 250nm were intercepted respectively. The results showed that the amount of information at the wavelength of 250nm was large, the chromatographic peak response value was moderate, and the baseline was relatively stable, so 250nm was selected as the detection wavelength.

[0129] ③Test sample preparation inspection

[0130] Through the investigation of the above-mentioned different preparation methods, it was found that there was almost no chromatographic peak in ethyl acetate ultrasonic treatment; the number of chromatographic peaks in ethanol ultrasonic treatment and n-butanol ultrasonic treatment was less and the response value was lower; compared with methanol, 50% methanol and 80% methanol ultrasonic extraction, there was no significant difference in the number of chromatographic peaks, but the response value of methanol ultrasonic extraction was higher. Finally, it was determined that the preparation method of the test sample was methanol ultrasonic treatment.

[0131] ④ Attribution of medicinal flavor peaks

[0132] The determination was carried out according to the determined chromatographic conditions to determine the medicinal taste to which each chromatographic peak in the characteristic spectrum of the composite granules belongs. The results are shown in Table 2.

[0133] Table 2 Chromatographic peaks attribution of characteristic spectrum of composite granules

[0134]

[0135]

[0136] According to the test results, chromatographic peaks 1, 3, 4, 6, and 8-21 are exclusive peaks of the prescription medicinal flavors. Peaks 1, 3, 6, and 8 belong to white peony root, peak 4 belongs to lily, peaks 9, 10, 11, 13, 14, 15, 17, and 18 belong to polygala root, peaks 12 and 16 belong to tangerine peel, peak 19 belongs to licorice root, peak 20 belongs to bupleurum, and peak 21 belongs to schisandra. Chromatographic peak 2 is a common peak of tangerine peel and schisandra; chromatographic peak 5 is a common peak of bupleurum, polygala root, and lily; chromatographic peak 7 is a common peak of tangerine peel and licorice root.

[0137] ⑤Chromatographic peak identification

[0138] UPLC-Q-TOF-MS was used to qualitatively analyze the chemical components of the composite granules, and the chromatographic peaks of the characteristic spectrum were identified and speculated. According to the UPLC-Q-TOF-MS analysis results, the reference HPLC method was used for further verification, and the chromatographic peaks were identified to obtain the test results. In the characteristic spectrum of the composite granules, each chromatographic peak corresponds to the reference substance: peak 1-gallic acid, peak 7-peony lactone glycosides, peak 8-peony glycosides, peak 11-polygala xanthophylloside III, peak 12-naringin, peak 13-3,6-dierucyl sucrose, peak 18-benzoylpeony glycosides, peak 20-saponin b1, peak 21-ammonium glycyrrhizate.

[0139] Table 3 Mass spectrometry analysis results of composite granules

[0140]

[0141]

[0142] Table 4 Correspondence between the classification of medicinal flavors and the results of characteristic peak identification

[0143]

[0144] Table 5 Chromatographic peak identification of characteristic spectrum of composite granules

[0145]

[0146]

[0147] ⑥ Reference object selection

[0148] After comparison with the reference substance, the chromatographic peak at 33.820 min was naringin, among which the chromatographic peak separation of naringin was better, the chromatographic peak shape was better, and the retention time was moderate, so it was calibrated as the S peak, and the naringin reference substance was used as a reference.

[0149] (2) Preliminary determination of the characteristic map method

[0150] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile was used as mobile phase A, 0.15% phosphoric acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 1 ml per minute; the column temperature was 30°C; the detection wavelength was 250 nm. The theoretical plate number calculated based on the naringin peak should be no less than 5000.

[0151] Table 6 Gradient elution table

[0152]

[0153]

[0154] Preparation of reference solution: Take an appropriate amount of naringin reference substance, weigh it accurately, and add methanol to make a solution containing 0.1 mg per 1 ml.

[0155] Preparation of test solution: Grind the product into powder, take about 1.5 g, weigh accurately, put it in a stoppered conical flask, add 25 ml of methanol accurately, stopper it, weigh it, and treat it with ultrasound (power 480 W, frequency 40 kHz) for 30 minutes. Take it out, cool it, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain it.

[0156] Determination method: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, measure, and record the chromatogram.

[0157] Methodological validation

[0158] ① Specificity and integrity inspection

[0159] Take this product (batch number: 240701), grind it into powder, take about 1.5g, put it in a stoppered conical flask, and operate according to the preparation and determination method of the test solution under the "Preliminary determination of the characteristic spectrum method". The collection method is extended by 30 minutes. Blank solvent, excipients, reference materials, and test samples are collected separately to investigate the specificity of the characteristic spectrum determination method.

[0160] Result analysis: The blank solvent, test solution, excipient solution and reference solution were measured respectively. The results showed that the blank solvent and excipient solution had no interference and the specificity was good.

[0161] ②Instrument precision test

[0162] Take this product (batch number: 240701), grind it into powder, take about 1.5g, weigh it accurately, put it in a stoppered conical flask, and operate according to the preparation and determination method of the test solution under the "Preliminary determination of the characteristic spectrum method", inject the sample 6 times continuously, and calculate the relative retention time and relative peak area of ​​each chromatographic peak with S peak as reference.

[0163] Table 7 Relative retention time results of instrument precision test (n=6)

[0164]

[0165] The results showed that the relative retention time RSD was less than 2.0%, indicating that the instrument had good precision.

[0166] ③Stability test

[0167] Take this product (batch number: 240701), grind it into powder, take about 1.5 g, weigh it accurately, and place it in a stoppered conical flask. According to the preparation and determination method of the test solution under the "Preliminary determination of the characteristic spectrum method", the sample is injected and measured at 0, 2, 4, 8, 12, 24, 36, and 48 hours respectively. Taking the S peak as a reference, calculate the relative retention time of each chromatographic peak. The specific determination results are shown in Table 8.

[0168] Table 8 Relative retention time results of stability test (n=8)

[0169]

[0170] The results showed that the relative retention time RSD was <2.0%, indicating that the test solution had good stability within 48 hours.

[0171] ④Repeatability test

[0172] Take this product (batch number: 240701), grind it into powder, take about 1.5 g, weigh it accurately, and place it in a stoppered conical flask. Follow the preparation and determination method of the test solution under the "Preliminary determination of the characteristic spectrum method" and calculate the relative retention time and relative peak area of ​​each chromatographic peak with the S peak as a reference. The specific determination results are shown in Table 9.

[0173] Table 9 Relative retention time results of repeatability test (n=6)

[0174]

[0175]

[0176] The results showed that the relative retention time RSD was less than 2.0%, indicating that the method had good repeatability.

[0177] ⑥Durability test

[0178] The durability test was conducted to investigate the effects of different flow rates, column temperatures, chromatographic columns, and acid concentrations on the chromatographic conditions. The relative retention times of the remaining peaks were calculated with the S peak as a reference. The specific measurement results are shown in Table 10.

[0179] Table 10 Test results at different flow rates

[0180]

[0181]

[0182] Table 11 Test results at different column temperatures

[0183]

[0184] Table 12 Test results of different chromatographic columns

[0185]

[0186]

[0187] Table 13 Test results of different acid concentrations

[0188]

[0189] The results showed that the determination results under different durability conditions were basically consistent, with 15 characteristic peaks appearing in the chromatograms and the relative retention times being basically consistent, indicating that the method had good durability.

[0190] (3) Determination of characteristic spectrum measurement method

[0191] Determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 General Rules 0512)

[0192] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile was used as mobile phase A, 0.15% phosphoric acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; the flow rate was 1 ml per minute; the column temperature was 30°C; the detection wavelength was 250 nm. The theoretical plate number calculated based on the naringin peak should be no less than 5000.

[0193] Table 14 Gradient elution table

[0194]

[0195] Preparation of reference solution: Take an appropriate amount of naringin reference substance, weigh it accurately, and add methanol to make a solution containing 0.1 mg per 1 ml.

[0196] Preparation of test solution: Grind the product into powder, take about 1.5 g, weigh accurately, put it in a stoppered conical flask, add 25 ml of methanol accurately, stopper it, weigh it, and treat it with ultrasound (power 480 W, frequency 40 kHz) for 30 minutes. Take it out, cool it, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain it.

[0197] Determination method: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, measure, and record the chromatogram.

[0198] (4) Establishment of reference characteristic map

[0199] Using the above-determined method, 12 batches of samples were measured, and the characteristic spectra of the 12 batches of samples were analyzed. The chromatographic peaks with good stability and appropriate response values ​​were selected as characteristic peaks, and a total of 15 common peaks were calibrated.

[0200] Summary: According to the absorption characteristics of the active ingredients of each medicinal ingredient in the prescription, the characteristic spectrum of the oral solid preparation of traditional Chinese medicine for the treatment of depression was established by HPLC. There are 15 characteristic peaks with strong specificity. A comprehensive study was made on the attribution of each characteristic peak, and each peak was attributed to each medicinal material. It can better control the quality of Bupleurum, White Peony Root, Polygala, Poria, Citrus aurantium, Albizzia julibrissin, Lilium, Schisandra chinensis, and Glycyrrhiza uralensis in the prescription; 15 indicator components were identified, which can more comprehensively control the main ingredients in the prescription, making the product quality more stable and controllable.

[0201] 3. Technical Effect

[0202] Before formal modeling and drug administration, rats in each group were adaptively raised in a rat breeding room for 1 week, and the status of rats in each group was observed. In the formal experiment, SD rats were divided into a blank group and a chronic unpredictable stress (CUMS) depression model group.

[0203] Except for the blank group, the rats in the CUMS group were exposed to alternating stressors every day for 39 consecutive days. After the modeling, sucrose preference tests and open field tests were performed. The sucrose preference rate and the number of standing, crossing grids, and the total number of crossing grids and standing in the open field test were used to evaluate whether the modeling of the rats in the CUMS group was successful. During the modeling period, each group of rats was only given normal diet and drinking water, and no corresponding drug treatment was given. After the modeling was successful, according to the results of the open field test and the sucrose preference test, the CUMS group was divided into a model group, a paroxetine group, an embodiment 1 to 3 group, and a comparative example 1 to 9 group, with 12 rats in each group, to ensure that there was no significant difference in the performance in the behavioral test, thereby ensuring the scientificity and accuracy of the subsequent experiments. The drug was administered continuously for 32 days, the paroxetine group was administered at a dose of 3 mg / kg body weight, and the embodiment 1 to 3 group and the comparative example 1 to 9 group were administered at a dose of 960 mg / kg body weight (1g of the composition granules corresponds to 18g of raw medicinal materials). Dissolve it in distilled water to prepare the required concentration before use. The animals in the normal group and the model group were gavaged with the same volume of distilled water.

[0204] Modeling method: fasting and water deprivation for the whole day, day and night reversal, hot and humid food, body restraint for 2 hours, 4℃ ice water bath for 5 minutes, tail clamping for 3 minutes, strobe, noise, one or two stimulation methods per day, the same stimulation method will not be repeated within a week to prevent the rats from predicting the upcoming stimulation method and affecting the modeling effect. See Table 15 for specific operations.

[0205] Table 15 Chronic unpredictable stress stimulation methods and operation steps for rats

[0206]

[0207]

[0208] 1. Effect of the combined granules on serum ACTH, CORT and CRH levels in CUMS rats

[0209] Table 16 Effect of the combined granules on serum ACTH, CORT and CRH levels in CUMS rats (x±s)

[0210]

[0211] 2. Effect of the combined granules on the content of 5HT and its metabolite 5H IAA in the cortex

[0212] Table 17 Effect of the combined granules on the content of 5-HT and 5-HIAA in rat cortex (x±s)

[0213]

[0214]

[0215] Conclusion: In the CUMS model, the combined granules have an antidepressant effect, and the mechanism of action is mainly related to inhibiting the excessive hyperactivity of the HPA axis under stress, improving the function of 5HT neurons and regulating neuronal plasticity.

[0216] The links not described in detail in the present invention are all common knowledge that can be selected by ordinary technicians in the field. Although the present invention has been described in detail above with general descriptions and specific implementation schemes, it is obvious to ordinary technicians in the field that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart, characterized in that: The Chinese medicine composition is composed of the following components: bupleurum, white peony root, polygala, tuckahoe, red tangerine peel, jujube bark, lily, schisandra chinensis, and roasted liquorice.

2. The Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition formula consists of the following components in parts by weight: 2 parts of Bupleurum chinense, 3 parts of White Peony Root, 2 parts of Polygala tenuifolia, 2 parts of Poria cocos, 2 parts of Citrus aurantium, 2 parts of Albizzia julibrissin bark, 2 parts of Lilium bulbs, 1 part of Schisandra chinensis, and 1 part of Radix Glycyrrhizae Preparata.

3. A Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart, characterized in that: The Chinese medicine composition is composed of the following components: water extracts of white peony root, tuckahoe, red tangerine peel, julibrissin bark, lily, and roasted liquorice root, alcohol extracts of bupleurum and schisandra chinensis, and enzymatic acid water extracts of bupleurum and schisandra chinensis residues and polygala root slices.

4. The Chinese medicine composition according to claim 3, characterized in that: The Chinese medicine composition is obtained from the following raw materials in parts by weight: 2 parts of bupleurum, 3 parts of white peony root, 2 parts of polygala, 2 parts of tuckahoe, 2 parts of tangerine peel, 2 parts of julibrissin bark, 2 parts of lily, 1 part of schisandra chinensis, and 1 part of roasted liquorice.

5. A method for preparing a Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart, characterized in that: The preparation method comprises the following steps: White peony root, Poria cocos, tangerine peel, Albizzia julibrissin, lily, and licorice root are extracted with water, and the liquid is dried to obtain a water extract of white peony root, Poria cocos, tangerine peel, Albizzia julibrissin, lily, and licorice root; Bupleurum root and Schisandra chinensis are extracted by reflux, and the liquid is dried to obtain a Bupleurum root and Schisandra chinensis alcohol extract; Bupleurum root and Schisandra chinensis residues and coarse powder of Polygala tenuifolia are hydrolyzed with water, then boiled and extracted, and the liquid is dried to obtain a hydrolyzed acid water extract of Bupleurum root and Schisandra chinensis residues and Polygala tenuifolia root; The water extracts of white peony root, tuckahoe, red tangerine peel, julibrissin bark, lily and roasted liquorice root, the alcohol extracts of bupleurum root and schisandra chinensis, and the enzymatic acid water extracts of bupleurum root and schisandra chinensis residues and polygala root slices are mixed to obtain a composition.

6. The method for preparing the Chinese medicine composition according to claim 5, characterized in that: The specific steps of the preparation method are: Add 8-10 times the amount of water to the white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root slices, decoct for 60 minutes, filter, add 6-8 times the amount of water to the residue, decoct for 30 minutes, filter, concentrate and dry the combined medicinal liquid, grind the dry paste and pass through a 60-mesh sieve to obtain the white peony root, Poria cocos, red tangerine peel, Albizzia julibrissin, lily and licorice root water extract; The crude powder of the bupleurum and schisandra pieces is refluxed and extracted twice, and 7 to 9 times the amount of 85 to 95% ethanol is added for the first time, extracted for 1 hour, filtered, and 6 to 8 times the amount of 85 to 95% ethanol is added for the second time, extracted for 1 hour, filtered, and the combined medicinal solution is concentrated under reduced pressure at 50 to 70° C. and dried, and the dry paste is crushed and passed through a 60-mesh sieve to obtain the bupleurum and schisandra alcohol extract; The dregs of Radix Bupleuri and Schisandrae Chinensis and the coarse powder of Radix Polygalae are extracted with water twice, the first time the amount is 8 to 12 times, the pH value is adjusted to 4.3 to 5.3, 1000 to 3000 U of pectinase and 1000 to 2000 U of cellulase are added per kilogram of the decoction pieces, the enzymatic hydrolysis is carried out at 40 to 50° C. for 5 to 7 hours, and then the extraction is boiled for 1 hour, and the extraction is centrifuged, the second time the amount is 7 to 9 times, the pH value is adjusted to 4.3 to 5.3, the extraction is boiled for 1 hour, and the extraction is centrifuged, the combined medicinal solution is concentrated under reduced pressure at 60 to 90° C. and dried, the dry paste is crushed and passed through a 60-mesh sieve, and the enzymatic acid water extract of the dregs of Radix Bupleuri and Schisandrae Chinensis and the decoction pieces of Radix Polygalae is obtained; The water extracts of white peony root, tuckahoe, red tangerine peel, julibrissin bark, lily and roasted liquorice root, the alcohol extracts of bupleurum root and schisandra chinensis, and the enzymatic acid water extracts of bupleurum root and schisandra chinensis residues and polygala root slices are mixed to obtain a composition.

7. Use of the composition according to any one of claims 3 and 4 in the preparation of a drug for treating depression.

8. A method for constructing a fingerprint spectrum of a traditional Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart, characterized in that: The steps include: Take gallic acid, astragaloside, paeoniflorin, lily A, paeoniflorin, sibirica polygala xanthoside A, polygala xanthoside III, naringin, 3,6'-diesinapoylsucrose, polygala tenuifolia glycoside A, licorice glycoside, benzoylpeoniflorin, ammonium glycyrrhizinate, saikosaponin b1, schisandrae alcohol A reference substances to prepare reference substance solution; Take a Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices, and prepare a test solution of the Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices; The test solution of the Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices and the reference solution are subjected to high performance liquid chromatography analysis respectively to construct a fingerprint of the Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices; The conditions for high performance liquid chromatography analysis include: acetonitrile as mobile phase A and 0.15% phosphoric acid solution as mobile phase B; gradient elution; the gradient elution program includes: 0min to 10min, the volume percentage of mobile phase A changes from 5% to 10%; 10min to 25min, the volume percentage of mobile phase A changes from 10% to 20%; 25min to 30min, the volume percentage of mobile phase A changes from 20% to 25%; 30min to 35min, the volume percentage of mobile phase A changes from 25% to 28%; 35min to 50min, the volume percentage of mobile phase A changes from 28% to 40%; 50min to 65min, the volume percentage of mobile phase A changes from 40% to 55%.

9. The method for constructing the fingerprint of the Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and orifices according to claim 8, characterized in that: The fingerprint of the traditional Chinese medicine composition for treating depression caused by liver qi stagnation and phlegm blocking the heart and mind comprises 15 common peaks; wherein, taking peak 8 naringin as the reference peak, the relative retention time of peak 1 gallic acid is 0.221±10%, the relative retention time of peak 2 astragaloside is 0.519±10%, the relative retention time of peak 3 paeoniflorin is 0.686±10%, the relative retention time of peak 4 royal lily A is 0.705±10%, the relative retention time of peak 5 paeoniflorin is 0.740±10%, the relative retention time of peak 6 Siberian polygala xanthoconone A is 0.814±10%, and the relative retention time of peak 7 polygala xanthoconone is 0.804±10%. The relative retention time of peak Ⅲ was 0.875±10%, the relative retention time of peak 9 for 3,6'-diesinapoylsucrose was 1.019±10%, the relative retention time of peak 10 for polygala tenuifolia glycoside A was 1.092±10%, the relative retention time of peak 11 for liquiritin was 1.128±10%, the relative retention time of peak 12 for benzoylpeonidin was 1.364±10%, the relative retention time of peak 13 for ammonium glycyrrhizinate was 1.669±10%, the relative retention time of peak 14 for saikosaponin b1 was 1.745±10%, and the relative retention time of peak 15 for schisandrin A was 1.782±10%.

10. An application of the fingerprint spectrum as claimed in claim 8 or 9 as a standard fingerprint spectrum in drug testing and quality control for the treatment of depression.

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