Ginseng and tortoise foundation strengthening wine as well as quality detection method and application thereof

Abstract

The invention discloses ginseng and tortoise root strengthening wine as well as a detection method and application thereof. The ginseng and tortoise root strengthening wine is prepared from angelica sinensis, astragalus membranaceus, codonopsis pilosula, poria cocos, dogwood, ligusticum wallichii, rhizoma atractylodis macrocephalae, schisandra chinensis, pericarpium citri reticulatae, fructus lycii, divaricate saposhnikovia roots, notopterygium roots, poria with hostwood, radix ophiopogonis, rehmannia, prepared rehmannia roots, Chinese dates and tortoise-shell glue. The traditional Chinese medicine composition adopts the theory of treatment based on syndrome differentiation to screen the composition of the traditional Chinese medicine raw materials, is scientific and reasonable in proportion, and is prepared into traditional Chinese medicine health care wine. Experimental results show that the ginseng and tortoise foundation strengthening wine provided by the invention has various remarkable effects of resisting aging, improving immunity, resisting inflammation, easing pain, relieving cough and reducing sputum, tranquilizing and allaying excitement, improving memory, strengthening physique, tonifying qi and activating blood and the like. In addition, the identification method and the content determination method provided by the invention can be used for comprehensively and objectively determining the content of the active ingredients, and have important significance for ensuring the clinical curative effect of the active ingredients.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine health-care wine, and in particular to a Shengui Guben wine having remarkable effects of supplementing qi and nourishing blood, strengthening the spleen and nourishing the kidney, dispelling dampness and activating collaterals, and its quality detection method and application. Background Art

[0002] Traditional Chinese medicine medicinal wines use white wine, yellow rice wine or rice wine as the base wine, soak or decoct specific traditional Chinese medicine formulas to dissolve the effective components of the medicine, and have both medicinal and wine effects. Wine can enhance the absorption of medicine and expand the scope of the medicinal effect. "Huangdi Neijing" records that "Laoli" (the prototype of medicinal wine) is used for treating diseases. The specific formula of medicinal wine is clearly stated in "Treatise on Febrile and Miscellaneous Diseases", such as "Danggui Sini Decoction with Evodia and Fresh Ginger" is decocted with wine for oral administration.

[0003] Traditional Chinese medicine medicinal wines have the effects of enhancing immunity, improving blood circulation, relieving joint pain, nourishing and strengthening the body, etc.

[0004] Based on the existing technology, the present invention prepares a traditional Chinese medicine medicinal wine with remarkable effects by screening out specific traditional Chinese medicine flavors and their ratios. Summary of the Invention

[0005] Object of the Invention: The object of the present invention is to solve the deficiencies of the existing technology and provide a Shengui Guben wine with a scientific and reasonable ratio, definite curative effect, and remarkable effects of supplementing qi and nourishing blood, strengthening the spleen and nourishing the kidney, dispelling dampness and activating collaterals, and its quality detection method and application.

[0006] Technical Solution: In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0007] A Shengui Guben wine, characterized in that it is made from the following raw materials:

[0008] Angelica sinensis, Astragalus membranaceus, Codonopsis pilosula, Poria cocos, Cornus officinalis, Ligusticum wallichii, Atractylodes macrocephala, Schisandra chinensis, Citrus reticulata Blanco, Lycium barbarum, Saposhnikovia divaricata, Notopterygium incisum, Poria cum Radix Pini, Ophiopogon japonicus, Rehmannia glutinosa Libosch., Rehmannia glutinosa (Gaertn.) DC., Jujube, Tortoise shell glue.

[0009] As a preferred embodiment, the above-mentioned Shengui Guben wine is made from the following raw materials in parts by weight:

[0010] Angelica sinensis 15 - 45 parts, Astragalus membranaceus 30 - 90 parts, Codonopsis pilosula 15 - 45 parts, Poria cocos 18 - 54 parts, Cornus officinalis 18 - 54 parts, Ligusticum wallichii 10 - 30 parts, Atractylodes macrocephala 30 - 90 parts, Schisandra chinensis 12 - 36 parts, Citrus reticulata Blanco 18 - 54 parts, Lycium barbarum 15 - 45 parts, Saposhnikovia divaricata 10 - 30 parts, Notopterygium incisum 10 - 30 parts, Poria cum Radix Pini 30 - 90 parts, Ophiopogon japonicus 7.5 - 22.5 parts, Rehmannia glutinosa Libosch. 35 - 105 parts, Rehmannia glutinosa (Gaertn.) DC. 30 - 90 parts, Jujube 35 - 105 parts, Tortoise shell glue 10 - 30 parts.

[0011] As a further preferred embodiment, the Shengui Guben Wine described above is made from the following raw materials in parts by weight:

[0012] Angelica sinensis 15 - 30 parts, Astragalus membranaceus 30 - 60 parts, Codonopsis pilosula 15 - 30 parts, Poria cocos 18 - 36 parts, Cornus officinalis 18 - 36 parts, Ligusticum chuanxiong 10 - 20 parts, Atractylodes macrocephala 30 - 60 parts, Schisandra chinensis 12 - 24 parts, Citrus reticulata Blanco 18 - 36 parts, Lycium barbarum 15 - 30 parts, Saposhnikovia divaricata 10 - 20 parts, Notopterygium incisum 10 - 20 parts, Poria cum Radix Pini 30 - 60 parts, Ophiopogon japonicus 7.5 - 15 parts, Rehmannia glutinosa Libosch. 35 - 70 parts, Rehmannia glutinosa (Gaertn.) Libosch. f. hueichingensis (Chao & Schih) Hsiao & Keng 30 - 60 parts, Ziziphus jujuba Mill. 35 - 70 parts, tortoise shell glue 10 - 20 parts.

[0013] As a particularly preferred embodiment, the Shengui Guben Wine described above is made from the following raw materials in parts by weight:

[0014] Angelica sinensis 15 parts, Astragalus membranaceus 31.25 parts, Codonopsis pilosula 15 parts, Poria cocos 18.75 parts, Cornus officinalis 18.75 parts, Ligusticum chuanxiong 11.25 parts, Atractylodes macrocephala 31.25 parts, Schisandra chinensis 12.5 parts, Citrus reticulata Blanco 18.75 parts, Lycium barbarum 15 parts, Saposhnikovia divaricata 11.25 parts, Notopterygium incisum 11.25 parts, Poria cum Radix Pini 31.25 parts, Ophiopogon japonicus 7.5 parts, Rehmannia glutinosa Libosch. 37.5 parts, Rehmannia glutinosa (Gaertn.) Libosch. f. hueichingensis (Chao & Schih) Hsiao & Keng 31.25 parts, Ziziphus jujuba Mill. 37.5 parts, tortoise shell glue 11.25 parts.

[0015] The preparation method of the Shengui Guben Wine described in the present invention includes the following steps:

[0016] Take Angelica sinensis, Astragalus membranaceus, Codonopsis pilosula, Poria cocos, Cornus officinalis, Ligusticum chuanxiong, Atractylodes macrocephala, Schisandra chinensis, Citrus reticulata Blanco, Lycium barbarum, Saposhnikovia divaricata, Notopterygium incisum, Poria cum Radix Pini, Ophiopogon japonicus, Rehmannia glutinosa Libosch., Rehmannia glutinosa (Gaertn.) Libosch. f. hueichingensis (Chao & Schih) Hsiao & Keng, Ziziphus jujuba Mill., tortoise shell glue in parts by weight, after crushing, add rock sugar, then add white liquor, soak tightly, steam, cool, place in a cellar, filter, dilute with water, centrifuge, filter, and obtain the product.

[0017] As a preferred embodiment, the preparation method of the Shengui Guben Wine described above includes the following steps:

[0018] Take Angelica sinensis, Astragalus membranaceus, Codonopsis pilosula, Poria cocos, Cornus officinalis, Ligusticum chuanxiong, Atractylodes macrocephala, Schisandra chinensis, Citrus reticulata Blanco, Lycium barbarum, Saposhnikovia divaricata, Notopterygium incisum, Poria cum Radix Pini, Ophiopogon japonicus, Rehmannia glutinosa, Rehmannia glutinosa (Gaertn.) Libosch. f. hueichingensis (Chao & Schih) Hsiao & Keng, Ziziphus jujuba Mill., tortoise shell glue in parts by weight, after crushing, add rock sugar, then add white liquor in an amount of 5 - 50 times the amount of the medicinal materials, soak tightly for 24 - 48 hours, steam for 2 - 3 hours, cool, place in a cellar for 10 - 21 days, filter, dilute with water, centrifuge, filter, and obtain the product.

[0019] The identification method of the Shengui Guben Wine described in the present invention includes the following steps:

[0020] (1) Identification of control medicinal materials of Angelica sinensis and Ligusticum chuanxiong

[0021] (1.1) Take the Shengui Guben Wine, steam it until there is no alcohol smell, let it cool, transfer it to a separatory funnel in portions with water, shake and extract with ether, combine the extraction solutions, evaporate to dryness, dissolve the residue in ethyl acetate to obtain the test solution.

[0022] (1.2) Separately take the reference crude drugs of Angelica sinensis and Ligusticum chuanxiong, add ether to each, soak them separately in the cold, shake constantly, filter, evaporate the filtrate to dryness, dissolve the residue in ethyl acetate separately to obtain the reference crude drug solutions.

[0023] (1.3) Respectively absorb the test solution and the reference crude drug solutions in step (1), spot them on the same silica gel G thin-layer plate, use petroleum ether - ethyl acetate as the developing agent, develop, take out, dry in air, and examine under an ultraviolet lamp at 365 nm. In the test solution chromatogram, at the positions corresponding to the reference crude drug chromatogram, fluorescent spots of the same color are shown.

[0024] (2) Identification of loganin in Cornus officinalis

[0025] (2.1) Take the Shengui Guben Wine, steam it until nearly dry, dissolve it in ethyl acetate, heat under reflux, filter, evaporate the filtrate to dryness, dissolve the residue in methanol to obtain the test solution.

[0026] (2.2) Separately take the loganin reference substance, dissolve it in methanol to prepare the reference substance solution.

[0027] (2.3) Respectively absorb the test solution in step (2.1) and the reference substance in step (2.2), spot them on the same silica gel G thin-layer plate, use ethyl acetate - acetone - formic acid - water as the developing agent, develop, take out, dry in air, spray with 10% sulfuric acid ethanol solution, and bake until the spots are clearly developed; in the test solution chromatogram, at the positions corresponding to the reference substance chromatogram, spots of the same color are shown.

[0028] (3) Identification of deoxyschizandrin in Schisandra chinensis

[0029] (3.1) Take the Shengui Guben Wine, steam it until there is no alcohol smell, let it cool, transfer it to a separatory funnel in portions with water, extract with ethyl acetate, combine the extraction solutions, concentrate on a water bath to obtain the test solution.

[0030] (3.2) Separately take the deoxyschizandrin reference substance, dissolve it in ethyl acetate to prepare the reference substance solution.

[0031] (3.3) Absorb the test solution in step (3.1) and the reference substance in step (3.2), spot them on the same silica gel GF254 thin-layer plate, use petroleum ether - ethyl acetate - formic acid as the developing agent, develop, take out, dry in air, and examine under an ultraviolet lamp at 254 nm. In the test solution chromatogram, at the positions corresponding to the reference substance chromatogram, spots of the same color are shown.

[0032] (4) Identification of the control crude drug of Chinese wolfberry fruit

[0033] (4.1) Take Shengui Guben Wine, steam it until there is no alcohol smell, let it cool, transfer it to a separating funnel in portions with water, extract with ethyl acetate, combine the extraction solutions, concentrate them on a water bath, and use it as the test solution;

[0034] (4.2) Take the control crude drug of Chinese wolfberry fruit, warm-soak it with ethyl acetate, filter, concentrate the filtrate, and use it as the control crude drug solution;

[0035] (4.3) Take the test solution and the control crude drug solution, spot them on the same silica gel G thin layer plate respectively, use ethyl acetate - trichloromethane - formic acid as the developing agent, develop, take out, dry in the air, and examine under ultraviolet light at 365 nm; in the test solution chromatogram, at the position corresponding to the control crude drug chromatogram of Chinese wolfberry fruit, there are fluorescent spots showing the same color;

[0036] (5) Identification of hesperidin in tangerine peel

[0037] (5.1) Take Shengui Guben Wine, steam it until there is no alcohol smell, add water in portions and transfer it to a separating funnel, extract with n-butanol saturated with water, combine the extraction solutions, evaporate to dryness, dissolve the residue in methanol, and use it as the test solution;

[0038] (5.2) Separately take the reference substance of hesperidin, dissolve it in methanol to prepare a reference substance solution;

[0039] (5.3) Take the control crude drug of tangerine peel, ultrasonicate it with methanol, filter, concentrate the filtrate, and use it as the control crude drug solution;

[0040] (5.4) Absorb the test solution, the reference substance and the control crude drug solution, spot them on the same silica gel G thin layer plate respectively, use ethyl acetate - methanol - water as the developing agent, develop, take out, dry in the air, then use the upper layer solution of toluene - ethyl acetate - formic acid - water as the developing agent, develop, take out, dry in the air, spray with aluminum trichloride test solution, and examine under ultraviolet light at 365 nm. In the test solution chromatogram, at the positions corresponding to the control crude drug and the reference substance chromatograms, there are fluorescent spots showing the same color;

[0041] (6) Identification of oleanolic acid

[0042] (6.1) Take Shengui Guben Wine, steam it until there is no alcohol smell, let it cool, transfer it to a separating funnel in portions with water, shake and extract with ether, combine the extraction solutions, evaporate the solvent, dissolve the residue in ethyl acetate, and use it as the test solution;

[0043] (6.2) Take the reference substance of oleanolic acid, dissolve it in ethyl acetate to prepare a reference substance solution;

[0044] (6.3) Pipette the test solution and oleanolic acid reference solution and spot them on the same silica gel G thin layer plate respectively. Use toluene-ethyl acetate-glacial acetic acid as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and blow with hot air until the spots are clearly colored. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the reference.

[0045] As a preferred embodiment, the identification method of the above-mentioned Shengui Guben wine comprises the following steps:

[0046] (1) Identification of Chinese Angelica and Chuanxiong as control herbs

[0047] (1.1) Take Shengui Guben wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, and extract it 2 to 3 times with ether, 20 to 30 ml each time. Combine the extracts, evaporate them to dryness, and dissolve the residue with ethyl acetate to prepare the test solution;

[0048] (1.2) Take another reference drug, Angelica sinensis and Ligusticum chuanxiong, add ether to each, soak in cold water, shake from time to time, filter, evaporate the filtrate, and dissolve the residue in ethyl acetate to prepare the reference drug solution;

[0049] (1.3) The test solution and the control medicinal material solution of step (1) are respectively aspirated and spotted on the same silica gel G thin layer plate, and petroleum ether-ethyl acetate in a volume ratio of 20:1 is used as a developing solvent, developed, taken out, dried, and examined under a UV lamp at 365 nm. In the chromatogram of the test sample, a fluorescent spot of the same color is displayed at the corresponding position of the chromatogram of the control medicinal material;

[0050] (2) Identification of Loganin from Cornus officinalis

[0051] (2.1) Take ginseng and turtle wine, evaporate it to near dryness, add ethyl acetate to dissolve it, heat and reflux for 1 to 2 hours, filter it, evaporate the filtrate to dryness, and add methanol to dissolve the residue as the test solution;

[0052] (2.2) Take another loganin reference substance and add methanol to make a solution containing 1-5 mg of the reference substance per 1 ml;

[0053] (2.3) The test solution of step (2.1) and the reference substance of step (2.2) are respectively aspirated and spotted on the same silica gel G thin layer plate, and developed with ethyl acetate-acetone-formic acid-water in a volume ratio of 12:8:1:1, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and baked at 10-50° C. until the spots are clearly colored; in the chromatogram of the test sample, spots of the same color are displayed at the corresponding positions in the chromatogram of the reference substance;

[0054] (3) Identification of schisandrin A and schisandrin B in Schisandra chinensis

[0055] (3.1) Take the Shengui Guben Wine, steam it until there is no alcohol smell, let it cool, transfer it to a separating funnel in portions with water, extract with ethyl acetate 2 - 3 times, 20 - 30 ml each time, combine the extracts, concentrate on a water bath, and use it as the test solution;

[0056] (3.2) Separately take reference substances of schisandrin A and schisandrin B, and prepare a mixed reference substance solution containing 1 - 5 mg per 1 ml with ethyl acetate;

[0057] (3.3) Pipette the test solution from step (3.1) and the reference substance solution from step (3.2) respectively onto the same silica gel GF254 thin layer plate, use petroleum ether - ethyl acetate - formic acid with a volume ratio of 15:5:1 as the developing agent, develop, take out, dry in air, and examine under an ultraviolet lamp at 254 nm. In the test solution chromatogram, at the positions corresponding to the reference substance chromatogram, spots of the same color are shown.

[0058] (4) Identification of the control crude drug of wolfberry fruit

[0059] (4.1) Take the Shengui Guben Wine, steam it until there is no alcohol smell, let it cool, transfer it to a separating funnel in portions with water, extract with ethyl acetate 2 - 3 times, 20 - 30 ml each time, combine the extracts, concentrate on a water bath, and use it as the test solution;

[0060] (4.2) Take the control crude drug of wolfberry fruit, warm - soak it with ethyl acetate for 1 - 2 hours, filter, concentrate the filtrate, and use it as the control crude drug solution;

[0061] (4.3) Take the test solution and the control crude drug solution, respectively spot them on the same silica gel G thin layer plate, use ethyl acetate - chloroform - formic acid with a volume ratio of 3:2:1 as the developing agent, develop, take out, dry in air, and examine under an ultraviolet lamp at 365 nm; in the test solution chromatogram, at the positions corresponding to the control crude drug chromatogram of wolfberry fruit, fluorescent spots of the same color are shown;

[0062] (5) Identification of hesperidin in tangerine peel

[0063] (5.1) Take the Shengui Guben Wine, steam it until there is no alcohol smell, transfer it to a separating funnel in portions with water, extract with n - butanol saturated with water 1 - 3 times, 10 ml each time, combine the extracts, evaporate to dryness, dissolve the residue with methanol, and use it as the test solution;

[0064] (5.2) Separately take a reference substance of hesperidin, and prepare a reference substance solution with methanol;

[0065] (5.3) Then take the control crude drug of tangerine peel, ultrasonicate it with methanol, filter, concentrate the filtrate, and use it as the control crude drug solution;

[0066] (5.4) Pipette the test solution, reference substance and reference medicinal material solution, spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-water with a volume ratio of 100:17:13 as the developing solvent, develop, take out, dry, then use the upper layer solution of toluene-ethyl acetate-formic acid-water with a volume ratio of 20:10:1:1 as the developing solvent, develop, take out, dry, spray with aluminum chloride test solution, and examine under ultraviolet light at 365nm. In the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference medicinal material and the reference substance, a fluorescent spot of the same color is displayed;

[0067] (6) Identification of oleanolic acid

[0068] (6.1) Take the ginseng and turtle wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, and extract it 2 to 3 times with ether, 20 to 30 ml each time. Combine the extracts, evaporate them to dryness, and dissolve the residue with ethyl acetate to prepare the test solution;

[0069] (6.2) Take the oleanolic acid reference substance and add ethyl acetate to prepare a reference substance solution containing 1 to 5 mg per 1 ml;

[0070] (6.3) Pipette the test sample solution and oleanolic acid reference solution and spot them on the same silica gel G thin layer plate respectively. Use toluene-ethyl acetate-acetic acid in a volume ratio of 14:4:0.5 as the developing solvent. Develop the plate, take it out, dry it, spray it with 10% sulfuric acid ethanol solution, and blow it with hot air until the spots are clearly colored. In the chromatogram of the test sample, a spot of the same color appears at the corresponding position in the chromatogram of the reference.

[0071] The quality detection method of the ginseng and turtle guben wine of the present invention comprises the following steps:

[0072] (1) Preparation of reference solution

[0073] Take reference substance solutions of astragaloside IV, hesperidin, loganin, ferulic acid, oleanolic acid, schisandrin A and schisandrin B respectively, add ethanol solvent to prepare mixed reference substance solution;

[0074] (2) Take the ginseng and turtle wine, evaporate it to dryness, add ethanol to ultrasonically extract it, filter the extract, and filter it through a 0.22 μm filter membrane to use as the test solution;

[0075] (3) Establishment of standard curve

[0076] The mixed reference solution of step (1) was taken and diluted into 5 concentrations in sequence, and filtered through a 0.22 μm filter membrane respectively. 5 μL of the solution was taken and injected into HPLC for analysis, and the standard curve equation was drawn with the concentration as the horizontal axis and the peak area as the vertical axis;

[0077] (4) Inject 10 μL of the test solution obtained in step (2) into the HPLC for injection analysis to obtain a chromatogram. According to the retention time of the reference substance, substitute the peak area into the standard curve equation to calculate the content of each component.

[0078] As a preferred embodiment, in the quality inspection method of the Ginseng Turtle Consolidating the Root Wine described above, in step (1), take reference substances of astragaloside IV, hesperidin, loganin, ferulic acid, oleanolic acid, schisandrin A, and schisandrin B respectively, add ethanol solvent to prepare a mixed reference substance solution with concentrations of 2.0 mg / mL, 1.4 mg / mL, 0.62.0 mg / mL, 1.1 mg / mL, 0.50 mg / mL, 1.8 mg / mL, and 1.3 mg / mL respectively.

[0079] As a preferred embodiment, in the quality inspection method of the Ginseng Turtle Consolidating the Root Wine described above, in step (2), take 50 mL of the Ginseng Turtle Consolidating the Root Wine, evaporate to dryness, add 10 - 20 mL of ethanol and ultrasonically extract for 10 - 30 min, filter to obtain an extract, and pass through a 0.22 μm filter membrane as the test solution.

[0080] As a preferred embodiment, in the quality inspection method of the Ginseng Turtle Consolidating the Root Wine described above, the chromatographic conditions for steps (3) and (4) are

[0081] Waters Sun Fire C18 chromatographic column, the specification of the chromatographic column is 250 mm × 4.6 mm, 5 μm; use methanol as phase A and 0.1% phosphoric acid water as phase B for gradient elution. From 0 to 6.8 min, 40% - 60% A; from 6.8 to 15 min, 60% - 75% A; from 15 to 26 min, 75% - 85% A; from 26 to 34 min, 85% - 40% A; from 34 to 50 min, 40% A; from 50 to 80 min, 60% A; flow rate: 1 mL / min, column temperature: 30 °C, detection wavelength: 245 nm.

[0082] As a preferred embodiment, in the quality inspection method of the Ginseng Turtle Consolidating the Root Wine described above, the standard curve equations of each active ingredient are as follows:

[0083] Compound Standard curve <![CDATA[R 2 > Astragaloside IV y = 527.14x + 1574.32 0.9994 Hesperidin y = 1027.26x + 1652.42 0.9992 Loganin y = 824.31x - 1145.35 0.9988 Ferulic acid y = 319.25x + 854.63 0.9992 Oleanolic acid y = 765.19x + 1031.46 0.9990 Schisandrin A y = 1094.12x - 2010.23 0.9989 Schisandrin B y = 1572.57x - 1782.41 0.9991 .

[0084] Beneficial effects: Based on the theory of traditional Chinese medicine, the present invention selects the composition of traditional Chinese medicine raw materials by syndrome differentiation and treatment theory for traditional Chinese medicine formula screening. The provided traditional Chinese medicine composition has a scientific and reasonable ratio and is prepared into a traditional Chinese medicine health wine. Experimental results show that the Ginseng Turtle Consolidating the Root Wine provided by the present invention has various effects such as significant anti - aging, improving immunity, anti - inflammation, analgesia, relieving cough and resolving phlegm, sedating and calming the mind, improving memory, enhancing physical fitness, replenishing qi and activating blood circulation, etc.

[0085] The identification method and content determination method provided by the present invention can comprehensively and objectively determine the content of active ingredients, which is of great significance for ensuring its clinical efficacy. Description of the Drawings

[0086] Figure 1 It is the identification TLC diagram of the control medicinal materials of Angelica sinensis and Ligusticum chuanxiong.

[0087] Figure 2 It is the identification TLC diagram of the control medicinal material of Lycium barbarum.

[0088] Figure 3 It is the identification TLC diagram of the control medicinal material of oleanolic acid.

[0089] Figure 4 It is the HPLC chromatogram of the reference substance solution.

[0090] Figure 5 It is the HPLC chromatogram of the test solution. Detailed Embodiments

[0091] The present invention will be further illustrated below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. After reading the present invention, various equivalent modifications made by those skilled in the art fall within the scope defined by the appended claims of this application.

[0092] Example 1

[0093] 1. A Shengui Guben Wine is made from the following raw materials in parts by weight:

[0094] Angelica sinensis 15g, Astragalus membranaceus 31.25g, Codonopsis pilosula 15g, Poria cocos 18.75g, Cornus officinalis 18.75g, Ligusticum chuanxiong 11.25g, Atractylodes macrocephala 31.25g, Schisandra chinensis 12.5g, Citrus reticulata Blanco 18.75g, Lycium barbarum 15g, Saposhnikovia divaricata 11.25g, Notopterygium incisum 11.25g, Poria cum Radix Pini 31.25g, Ophiopogon japonicus 7.5g, Rehmannia glutinosa Libosch. var. purpurea Makino 37.5g, Rehmannia glutinosa Libosch. 31.25g, Ziziphus jujuba Mill. 37.5g, Colla carapacis et plastri Testudinis 11.25g.

[0095] 2. The preparation method of the Shengui Guben Wine according to the present invention includes the following steps:

[0096] Take Angelica sinensis, Astragalus membranaceus, Codonopsis pilosula, Poria cocos, Cornus officinalis, Ligusticum chuanxiong, Atractylodes macrocephala, Schisandra chinensis, Citrus reticulata Blanco, Lycium barbarum, Saposhnikovia divaricata, Notopterygium incisum, Poria cum Radix Pini, Ophiopogon japonicus, Rehmannia glutinosa Libosch. var. purpurea Makino, Rehmannia glutinosa Libosch., Ziziphus jujuba Mill., Colla carapacis et plastri Testudinis according to the above parts by weight. After crushing, add rock sugar, and then add 20 times the volume of liquor based on the weight of the medicinal materials. Seal and soak for 48 hours, then cook for 2 hours, let it cool, place it in the cellar for 21 days, filter, dilute with water, centrifuge, and filter to obtain.

[0097] 3. The identification method of the above-mentioned Shengui Guben wine comprises the following steps:

[0098] (1) Identification of Chinese Angelica and Chuanxiong as control herbs

[0099] (1.1) Take 50 ml of Shengui Guben wine, steam until there is no alcohol smell, cool it, transfer it to a separatory funnel with 10 ml of water, and extract it with ether for 3 times, 30 ml each time. Combine the extracts, evaporate to dryness, and dissolve the residue with ethyl acetate to prepare the test solution;

[0100] (1.2) Take 5 g of Angelica sinensis and Ligusticum chuanxiong as control medicinal materials, add 100 mL of ether to each, soak for 1 hour, shake from time to time, filter, evaporate the filtrate, and add ethyl acetate to dissolve the residue to obtain Angelica sinensis and Ligusticum chuanxiong control medicinal material solutions;

[0101] (1.3) 5 μl of the test sample solution from step (1) was taken separately, and three points of the test sample were spotted in parallel, and 5 μl of the reference medicinal material solutions of Angelica sinensis and Ligusticum chuanxiong were spotted on the same silica gel G thin layer plate, and petroleum ether-ethyl acetate with a volume ratio of 20:1 was used as the developing solvent. The plate was developed, taken out, dried, and examined under a UV lamp at 365 nm. In the chromatogram of the test sample, a fluorescent spot of the same color was displayed at the corresponding position of the chromatogram of the reference medicinal material; Figure 1 .

[0102] (2) Identification of Loganin from Cornus officinalis

[0103] (2.1) Take 50 ml of Shengui Guben wine, evaporate it to near dryness, add 25 ml of ethyl acetate to dissolve it, heat and reflux for 2 hours, filter it, evaporate the filtrate to dryness, and add methanol to dissolve the residue as the test solution;

[0104] (2.2) Take another loganin reference substance and add methanol to make a solution containing 2 mg of the reference substance per 1 ml;

[0105] (2.3) 10 μl of the test solution of step (2.1) and 5 μl of the reference substance of step (2.2) are respectively taken and spotted on the same silica gel G thin layer plate, and ethyl acetate-acetone-formic acid-water with a volume ratio of 12:8:1:1 is used as a developing solvent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and baked at 10-50° C. until the spots are clearly colored; in the chromatogram of the test sample, spots of the same color are displayed at the corresponding positions in the chromatogram of the reference substance;

[0106] (3) Identification of schisandrin A and schisandrin B in Schisandra chinensis

[0107] (3.1) Take 50 ml of Shengui Guben wine, steam until there is no alcohol smell, cool it, transfer it to a separatory funnel with 10 ml of water in batches, extract it with ethyl acetate three times, 20 ml each time, combine the extracts, concentrate them on a water bath, and use them as the test solution;

[0108] (3.2) Separately take schisandrin and schisandrin B reference substances, and prepare a mixed reference substance solution containing 2 mg of schisandrin and schisandrin B per 1 ml with ethyl acetate.

[0109] (3.3) Pipette 10 μl of the test solution from step (3.1) and 4 μl of the reference substance from step (3.2), and spot them on the same silica gel GF254 thin layer plate respectively. Use petroleum ether - ethyl acetate - formic acid with a volume ratio of 15:5:1 as the developing agent, develop, take out, dry in the air, and examine under an ultraviolet lamp at 254 nm. In the test solution chromatogram, at the position corresponding to the reference substance chromatogram, spots of the same color are shown.

[0110] (4) Identification of the control crude drug of wolfberry fruit

[0111] (4.1) Take 50 ml of Shengui Guben Wine, steam it until there is no alcohol smell, let it cool, transfer it to a separatory funnel in portions with 10 ml of water, extract with ethyl acetate twice, 30 ml each time, combine the extraction solutions, concentrate on a water bath, and use it as the test solution.

[0112] (4.2) Take 2 g of the control crude drug of wolfberry fruit, warm - soak it with 20 ml of ethyl acetate for 2 hours, filter, and concentrate the filtrate to use as the control crude drug solution.

[0113] (4.3) Pipette 5 μl of the test solution and 5 μl of the control crude drug solution. Spot 3 parallel spots of the test solution on the same silica gel G thin layer plate respectively. Use ethyl acetate - chloroform - formic acid with a volume ratio of 3:2:1 as the developing agent, develop, take out, dry in the air, and examine under an ultraviolet lamp at 365 nm; in the test solution chromatogram, at the position corresponding to the control crude drug chromatogram of wolfberry fruit, fluorescent spots of the same color are shown; such as Figure 2 。

[0114] (5) Identification of hesperidin in tangerine peel

[0115] (5.1) Take 50 ml of Shengui Guben Wine, steam it until there is no alcohol smell, add 10 mL of water and transfer it to a separatory funnel in portions, extract with n - butanol saturated with water 3 times, 10 ml each time, combine the extraction solutions, evaporate to dryness, dissolve the residue with methanol, and use it as the test solution.

[0116] (5.2) Separately take hesperidin reference substance, and prepare a reference substance solution containing 1 mg per 1 ml with methanol.

[0117] (5.3) Take another 2 g of the control crude drug of tangerine peel, ultrasonicate it with 20 mL of methanol, filter, and concentrate the filtrate to use as the control crude drug solution.

[0118] (5.4) Pipette 10 μl of the test solution, 5 μl of the reference substance solution and 5 μl of the control crude drug solution, spot them respectively on the same silica gel G thin layer plate, use ethyl acetate - methanol - water with a volume ratio of 100:17:13 as the developing solvent, develop, take out, air dry, then use the upper layer solution of toluene - ethyl acetate - formic acid - water with a volume ratio of 20:10:1:1 as the developing solvent, develop, take out, air dry, spray with aluminum chloride test solution, and examine under ultraviolet lamp at 365 nm. In the chromatogram of the test solution, at the positions corresponding to the chromatograms of the control crude drug and the reference substance, fluorescent spots of the same color should be shown;

[0119] (6) Identification of oleanolic acid

[0120] (6.1) Take 50 ml of Shengui Guben Wine, steam it until there is no alcohol smell, cool it, transfer it to a separating funnel in portions with 10 ml of water, extract it 3 times with ether by shaking, 30 ml each time, combine the extraction solutions, evaporate to dryness, dissolve the residue in ethyl acetate to obtain the test solution;

[0121] (6.2) Take oleanolic acid reference substance, dissolve it in ethyl acetate to prepare a reference substance solution containing 1 mg per 1 ml;

[0122] (6.3) Pipette 10 μl of the test solution and 4 μl of the oleanolic acid reference substance solution, spot them respectively on the same silica gel G thin layer plate, use toluene - ethyl acetate - glacial acetic acid with a volume ratio of 14:4:0.5 as the developing solvent, develop, take out, air dry, spray with 10% sulfuric acid ethanol solution, heat with hot air until the spots are clearly developed. In the chromatogram of the test solution, at the position corresponding to the chromatogram of the reference substance, spots of the same color should be shown, as Figure 3 .

[0123] 4. The quality inspection method of the Shengui Guben Wine described in the present invention includes the following steps:

[0124] (1) Preparation of the reference substance solution

[0125] Respectively take astragaloside IV, hesperidin, loganin, ferulic acid, oleanolic acid, schisandrin A and schisandrin B reference substances, add ethanol solvent to prepare a mixed reference substance solution with concentrations of 1.0 mg / mL, 0.4 mg / mL, 0.6 mg / mL, 1.1 mg / mL, 0.50 mg / mL, 1.5 mg / mL and 1.3 mg / mL respectively;

[0126] (2) Take 5 batches of 50 mL of Shengui Guben Wine, evaporate to dryness, add 10 mL of ethanol and ultrasonically extract for 30 min, filter the ultrasonic extraction solution to obtain the extraction solution, and pass it through a 0.22 μm filter membrane to obtain the test solution;

[0127] (3) Establishment of the standard curve

[0128] Successively pipette 1 mL, 2 mL, 3 mL, 4 mL, and 5 mL of the mixed reference substance solution from step (1), make up the volume to 10 mL, dilute to 5 concentrations, filter through a 0.22 μm filter membrane successively, and pipette 5 μL each and inject into the HPLC for sample analysis to obtain the chromatogram of the reference substance, as Figure 4 , (chromatographic conditions: Waters Sun Fire C18 chromatographic column, column specification 250 mm × 4.6 mm, 5 μm; using methanol as phase A and 0.1% phosphoric acid water as phase B, gradient elution, 0 - 6.8 min, 40% - 60% A; 6.8 - 15 min, 60% - 75% A; 15 - 26 min, 75% - 85% A; 26 - 34 min, 85% - 40% A; 34 - 50 min, 40% A; 50 - 80 min, 60% A. Flow rate: 1 mL / min, column temperature: 30 °C, detection wavelength 245 nm), with concentration as the abscissa and peak area as the ordinate, plot the standard curve equation as shown in Table 1 below.

[0129] Table 1 The standard curve equations of each active ingredient are as follows:

[0130] Compound Standard curve <![CDATA[R 2 > Astragaloside IV y = 527.14x + 1574.32 0.9994 Hesperidin y = 1027.26x + 1652.42 0.9992 Loganin y = 824.31x - 1145.35 0.9988 Ferulic acid y = 319.25x + 854.63 0.9992 Oleanolic acid y = 765.19x + 1031.46 0.9990 Schisandrin A y = 1094.12x - 2010.23 0.9989 Schisandrin B y = 1572.57x - 1782.41 0.9991

[0131] (4) Pipette 10 μL of the test solution of 5 batches from step (2), inject into the HPLC for sample analysis under the same chromatographic conditions as in step (3) to obtain the chromatogram, as Figure 5 (Batch 20241104), according to the retention time of the reference substance, substitute the peak area into the standard curve equation to calculate the content of each component. As shown in Table 2 below.

[0132] Table 2 The content of active ingredients in 5 batches of Shengui Guben Wine

[0133]

[0134] Example 2

[0135] 1. A kind of Shengui Guben Wine, which is made from the following raw materials in parts by weight:

[0136] Angelica sinensis 30 g, Astragalus membranaceus 60 g, Codonopsis pilosula 30 g, Poria cocos 36 g, Cornus officinalis 36 g, Ligusticum chuanxiong 20 g, Atractylodes macrocephala 60 g, Schisandra chinensis 24 g, Citrus reticulata Blanco 36 g, Lycium barbarum 30 g, Saposhnikovia divaricata 20 g, Notopterygium incisum 20 g, Poria cum Radix Pini 60 g, Ophiopogon japonicus 15 g, Rehmannia glutinosa Libosch. var. purpurea (Makino) Hsiao et Keng 70 g, Rehmannia glutinosa Libosch. 60 g, Ziziphus jujuba Mill. 70 g, Turtle shell glue 20 g.

[0137] 2. The preparation method of the Shengui Guben Wine described in the present invention, which includes the following steps:

[0138] Take angelica, astragalus root, codonopsis pilosula, poria cocos, cornel, chuanxiong rhizome, atractylodes macrocephala, schisandra chinensis, tangerine peel, wolfberry fruit, divaricate saposhnikovia root, incised notopterygium root, poria with hostwood, dwarf lilyturf tuber, rehmannia root, prepared rehmannia root, Chinese date, and tortoise plastron glue according to the above weight parts. After pulverization, add rock sugar and then add liquor with a volume 30 times that of the weight of the medicinal materials. Seal and soak for 48 hours, then steam for 2 hours, let it cool, place it in a cellar for 21 days, filter, dilute with water, centrifuge, and filter to obtain the product.

[0139] The identification method and the content detection method are the same as those in Example 1.

[0140] Example 3

[0141] A Shengui Guben wine is made from the following raw materials in the following weight parts:

[0142] Angelica 45g, astragalus root 90g, codonopsis pilosula 45g, poria cocos 54g, cornel 54g, chuanxiong rhizome 30g, atractylodes macrocephala 90g, schisandra chinensis 36g, tangerine peel 54g, wolfberry fruit 45g, divaricate saposhnikovia root 30g, incised notopterygium root 30g, poria with hostwood 90g, dwarf lilyturf tuber 22.5g, rehmannia root 105g, prepared rehmannia root 90g, Chinese date 105g, and tortoise plastron glue 30g.

[0143] The preparation method of the Shengui Guben wine described in the present invention includes the following steps:

[0144] Take angelica, astragalus root, codonopsis pilosula, poria cocos, cornel, chuanxiong rhizome, atractylodes macrocephala, schisandra chinensis, tangerine peel, wolfberry fruit, divaricate saposhnikovia root, incised notopterygium root, poria with hostwood, dwarf lilyturf tuber, rehmannia root, prepared rehmannia root, Chinese date, and tortoise plastron glue according to the above weight parts. After pulverization, add rock sugar and then add liquor with a volume 40 times that of the weight of the medicinal materials. Seal and soak for 24 hours, then steam for 3 hours, let it cool, place it in a cellar for 15 days, filter, dilute with water, centrifuge, and filter to obtain the product.

[0145] The identification method and the content detection method are the same as those in Example 1.

[0146] Example 4 Functional Experiment

[0147] I. Effect of enhancing physical fitness

[0148] 1. Effect on the fatigue resistance of forced swimming in mice

[0149] Fifty ICR strain mice, 25 males and 25 females, weighing 20±2 g, were randomly divided into 5 groups: the high-dose and low-dose groups of the traditional Chinese medicine medicated wine prepared in Example 1, a saline control group, and a positive control group of gypenoside tablets (produced by Ankang Traditional Chinese Medicine Factory). All groups were administered by gavage at the doses shown in Table 1, once a day for three consecutive days, and the experiment was conducted 1 hour after the last administration. A pool with a diameter of 30 cm and a water depth of 29 cm was used as the swimming venue, and the water temperature was 15°C. During the experiment, the animals were grouped and placed in the water, and forced to swim. The time from placing the animal in the water to its death was recorded, with the mouse's body sinking and its nose and mouth submerged in the water without the ability to float as the indicator. The results are shown in Table 3.

[0150] Table 3 Effects of the traditional Chinese medicine medicated wine prepared in Example 1 on the anti-fatigue ability of mice in forced swimming X-±SD

[0151] Group Dose Survival time (s) P value Saline control group 10 212.5±48.86 — — Xi'an Tequ group 10 0.15 ml / 10 g 218.6±10.5 2.80 >0.05 Low-dose group 10 0.1 ml / 10 g 323.4±28.31 66.65 <0.01 High-dose group 10 0.2 ml / 10 g 453.3±110.31 113.18 <0.01 Gynostemma pentaphyllum group 10 0.2 ml / 10 g 548.6±88.70 154.40 <0.01 。

[0152] Table 3 results showed that the normal pressure anoxia tolerance time of the animals in the traditional Chinese medicine medicated wine group prepared in Example 1 was significantly prolonged (P<0.05, P<0.01), but the anoxia tolerance time of the animals in the Xiantequ group did not change significantly (P>0.05), indicating that the traditional Chinese medicine medicated wine prepared in Example 1 has the effect of enhancing the anoxia tolerance ability of animals.

[0153] II. Analgesic effect

[0154] 1. Hot plate method

[0155] Fifty female ICR strain mice, weighing 20±2 g, were randomly divided into 5 groups: a normal saline control group, the high-dose and low-dose groups of the traditional Chinese medicine medicated wine prepared in Example 1, and a positive control group of pethidine. The hot plate temperature was adjusted to 50±0.5°C, and the time from placing the animal in the hot plate to its licking foot reaction (pain latency) was recorded as the pain threshold index. It was measured twice continuously, and the average value was taken as the pain threshold before administration. Then, the animals were administered by gavage at the doses shown in Table 4 for 3 consecutive days. One hour after the last administration, the pethidine group was intraperitoneally injected with 0.15 ml / 10 g of 0.5% solution 1 hour before the experiment, and then the above process was repeated. The pain threshold increase rate was calculated using the following formula:

[0156]

[0157] Table 4 Effects of the traditional Chinese medicine medicated wine prepared in Example 1 on the pain threshold of mice X-±SD

[0158]

[0159] The results in Table 4 show that both the high dose and low dose of the traditional Chinese medicine medicated wine prepared in Example 1 can significantly increase the pain threshold of mice (P<0.01), indicating that the traditional Chinese medicine medicated wine prepared in Example 1 has an obvious analgesic effect.

[0160] 2. Writhing method

[0161] Take 50 ICR strain white mice, group and administer drugs according to the hot plate method test method. After 1 hour, intraperitoneally inject 0.1 ml / 10 g of 0.6% glacial acetic acid, and observe the number of writhing reactions of the animals within 20 minutes after injecting glacial acetic acid. The pethidine group conducts the experiment 30 minutes after intraperitoneal injection of the drug. The experimental results of each drug administration group are compared with the saline control group. The results are shown in Table 5.

[0162] Table 5 Effects of the traditional Chinese medicine medicated wine prepared in Example 1 on the writhing reaction of mice X-±SD

[0163]

[0164] The results in Table 5 show that the traditional Chinese medicine medicated wine prepared in Example 1 can significantly reduce the number of writhing reactions of animals within 20 minutes, indicating that it has an obvious analgesic effect.

[0165] III. Anti-inflammatory effect

[0166] 1. Mouse ear swelling method

[0167] Take 50 male ICR strain white mice with a body weight of 26±4 g, and group and administer drugs as before. After continuous drug administration for 3 days, conduct the experiment. Apply 0.05 ml of croton oil mixture to both sides of the left ear of each mouse. After half an hour, for the normal saline group, apply 0.05 ml of normal saline per animal. For the high dose and low dose groups of the traditional Chinese medicine medicated wine prepared in Example 1, apply its stock solution and 50% dilution respectively. The positive control group applies triamcinolone acetonide acetate ointment. After 4 hours, sacrifice the animals by dislocation, remove both ears along the ear root, use a 9 mm direct punch to take ear pieces from the auricle, weigh them with a torsion balance, and subtract the weight of the right ear from the weight of the left ear, which is the swelling degree of the auricle. The results are compared with the normal saline group, and Table 6 shows the results.

[0168] Table 6 Effects of the traditional Chinese medicine medicated wine prepared in Example 1 on the swelling degree of mouse ears X-±SD

[0169] Group Number of animals (pcs) Degree of ear swelling in mice Inhibition rate (%) P value Saline control group 10 2.8±0.8 0.00 Low-dose medicinal wine group 10 1.7±0.1 32.14 <0.01 High-dose medicinal wine group 10 0.7±0.1 75.00 <0.01 Triamcinolone acetonide group 10 0.2±0.1 92.86 <0.001

[0170] The results in Table 6 show that the traditional Chinese medicine medicated wine prepared in Example 1 can significantly reduce the swelling degree of mice, indicating that it has an obvious anti-inflammatory effect.

[0171] 2. Rat paw edema method

[0172] Take 50 healthy male Sprague-Dawley (SD) rats and randomly divide them into 5 groups: a normal saline group, high-dose and low-dose groups of the traditional Chinese medicine medicated wine prepared in Example 1, and a dexamethasone intraperitoneal injection group. After continuous intragastric administration for three days, 1 hour after the last administration and 30 minutes after intraperitoneal injection of dexamethasone, inflammation induction was started. At the left and right ankle joints of each rat, 0.05 ml of 1% carrageenan was subcutaneously injected. The circumferences around the left and right ankle joints of the rats were measured before inflammation induction and at 1, 2, 3, 4, and 6 hours after inflammation induction. The difference in the circumferences before and after inflammation induction was taken as the swelling degree, and it was compared between each administration group and the normal saline group. The results are shown in Table 7.

[0173] Table 7 Effects of the traditional Chinese medicine medicated wine prepared in Example 1 on the paw swelling degree of rats X-±SD

[0174]

[0175] *P<0.05, **P<0.01, ***P<0.001;

[0176] The results were obvious. After three days of intragastric administration of the traditional Chinese medicine medicated wine prepared in Example 1, the paw swelling degree of rats caused by carrageenan was significantly reduced, and there was a dose-dependent relationship. The effect became evident 1 hour after administration and lasted for about 4 hours. This indicates that the traditional Chinese medicine medicated wine prepared in Example 1 has an obvious effect of inhibiting carrageenan-induced inflammation.

[0177] 3. Effects on proliferative inflammation (rat cotton ball implantation method)

[0178] Healthy male rats were randomly divided into 5 groups and administered by intragastric administration or intraperitoneal injection. Normal saline and dexamethasone were used as negative and positive controls respectively. Under light ether anesthesia, two sterile cotton balls weighing 50 mg were implanted subcutaneously in the groin of the rats. Administration started on the day of surgery, once a day for 7 consecutive days, and the cotton balls were removed on the 8th day. They were dried at 60 °C for 12 hours and weighed, and the weight of the original cotton ball was subtracted. This was the weight of granulation hyperplasia. It was compared between the administration group and the normal saline group, and the results are shown in Table 8.

[0179] Table 8 Effects of the traditional Chinese medicine medicated wine prepared in Example 1 on proliferative inflammation in rats X-±SD

[0180] Group Number of animals Dose Granulation tissue (mg / 100 g body weight) Inhibition rate (%) P value Saline control group 10 — 158.1±50.5 0.00 Low-dose group 10 0.1 ml / 100 / g 34.90±11.2 77.93 <0.01 High-dose group 10 0.2 ml / 100 / g 68.33±16.8 56.78 <0.01 Dexamethasone group 10 0.4 mg / 100 / g 18.60±9.30 88.22 <0.001

[0181] As can be seen from Table 8, the weight of the granulation tissue stimulated by the cotton balls in the animals of the traditional Chinese medicine medicated wine group prepared in Example 1 was significantly reduced. This indicates that it has the effect of inhibiting proliferative inflammation.

[0182] IV. Effects of sedation, calming the mind, and promoting memory function

[0183] 1. Sedative effect (synergistic method with sodium pentobarbital)

[0184] ICR strain mice, weighing 22 ± 4 g, with an equal number of males and females, were randomly divided into 4 groups: saline + pentobarbital group, Xiantequ + pentobarbital group, and high-dose and low-dose traditional Chinese medicine medicinal wine prepared in Example 1 + sodium pentobarbital group. First, the animals in each administration group were continuously intragastrically administered for three days. One hour after the last administration, the saline group was intragastrically administered with saline, and then the animals in each group were intraperitoneally injected with 25 mg / kg of sodium pentobarbital. The animals whose righting reflex disappeared for 1 minute were observed as being in a sleeping state, and the incidence of sleep in each group of animals was calculated. The results are shown in Table 9.

[0185] Table 9 Effects of the traditional Chinese medicine medicinal wine prepared in Example 1 on the synergistic sleep-promoting effect of sodium pentobarbital X-±SD

[0186]

[0187] As can be seen from the results in Table 9, the sleep rate of the animals in the traditional Chinese medicine medicinal wine group prepared in Example 1 increased significantly, and there was a dose-dependent relationship, indicating that it has an obvious sedative and tranquilizing effect.

[0188] 2. Effects on learning and memory function ("Y" maze method)

[0189] Forty ICR strain mice weighing 20 ± 2 g and forty old ICR strain mice weighing 28 ± 2 g were respectively randomly divided into 4 groups. All were administered by gavage. (1) Determination of conditioned reflex learning and memory function: Continuous administration was carried out for 10 days for the experiment. The experimental mice performed one round of "Y" maze tests every day, with a stimulation voltage of 45 mV, for 10 consecutive times. The 2nd and 3rd safe areas alternated, and a red vial was used as the safe area signal. The correct number of times the animals entered the safe area was recorded, and the average of the 10 days was taken as the experimental result. Each administration group was compared with the saline group. The results are shown in Table 10. (2) Memory retention test for old mice. The aforementioned old mice were taken and continuously subjected to 15 "Y" maze stimulation trainings for three consecutive days, so that in each 15 stimulations, more than 12 times were correct. At the same time, intragastric administration was carried out. After 4 days of stopping the training, on the 7th day of administration, the previous experiment was repeated. The results are shown in Table 10.

[0190] Table 10 Effects of the traditional Chinese medicine medicinal wine prepared in Example 1 on memory ability and memory retention ability X-±SD

[0191]

[0192] As can be seen in Table 10, after 10 days of administration, the conditioned reflex learning and memory ability of the animals in the administration group to the "Y" maze stimulation was improved, and after 1 day of administration, the memory retention ability of the old mice was significantly improved (P < 0.05). It shows that the traditional Chinese medicine medicinal wine prepared in Example 1 has the effect of improving memory ability and memory retention ability.

[0193] V. Spleen-Strengthening and Appetite-Improving Effect (Method of Inducing Spleen Deficiency with Rhubarb Decoction)

[0194] Fifty healthy ICR mice, weighing 21±3 g, with half males and half females, were randomly divided into 5 groups. Using normal saline as the negative control and Sijunzi Decoction as the positive control, each mouse was intragastrically administered 1 ml / head of 100% rhubarb decoction once a day for 8 consecutive days. Then, the mice in the drug administration group were intragastrically administered the traditional Chinese medicine medicinal wine prepared in Example 1, the saline group was intragastrically administered normal saline, and the positive control group was intragastrically administered 0.2 ml / head of 25% Sijunzi Decoction decoction. The body weight, body temperature and all conditions of the animals were observed, such as hair, diet, defecation and urination, and activities, etc. for 7 consecutive days. It was found that after 8 days of intragastric administration of rhubarb decoction, the body weight of the animals decreased significantly, the body temperature dropped, and symptoms of spleen deficiency such as anorexia, withered and dull hair, reduced activities and fear of cold appeared. With the extension of the drug administration time, the general condition of the animals improved, the body weight and body temperature recovered, and there were significant differences compared with the saline group. The results are shown in Table 11.

[0195] Table 11 Effects of the traditional Chinese medicine medicinal wine prepared in Example 1 on the body weight and body temperature of mice with spleen deficiency syndrome X-±SD

[0196]

[0197] *P<0.05, **P<0.01;

[0198] As can be seen from the results in Table 11, the traditional Chinese medicine medicinal wine prepared in Example 1 significantly increased the body weight and body temperature of the animals with spleen deficiency syndrome and improved the symptoms, proving that it has the effect of invigorating the spleen and promoting appetite.

[0199] VI. Cough-Suppressing and Phlegm-Removing Effect

[0200] 1. Cough-Suppressing Effect (Method of Inducing Asthma in Mice with Ammonia Water)

[0201] Healthy ICR mice, weighing 20±2 g, with half males and half females, were grouped and administered as before. Using normal saline as the negative control and Kehaokuai (product of Dalian Jinyi Pharmaceutical Factory, 911027) as the positive control, after continuous administration for three days, the animals were placed in a 500-ml bell jar, and a cotton ball dropped with 0.2 ml of ammonia water was placed in the bell jar. The number of coughs of the mice within 2 minutes was observed. The results are shown in Table 12. The results in the table show that the number of coughs of the animals in the high-dose and low-dose groups of the traditional Chinese medicine medicinal wine prepared in Example 1 within 2 minutes decreased significantly. (P<0.05), and there were significant differences compared with the saline group. It shows that the traditional Chinese medicine medicinal wine prepared in Example 1 has a cough-suppressing effect.

[0202] Table 12 Effects of the traditional Chinese medicine medicinal wine prepared in Example 1 on ammonia water-induced cough in mice X-±SD

[0203] Group Number of animals Dose Number of coughs (times / 2 minutes) Inhibition rate P value Normal saline group 10 — 76±17 0.00 — Xi'an Tequ group 10 0.15 ml / 10 g 75±15 1.32 >0.05 Low-dose medicinal wine group 10 0.1 ml / 10 g 51±13 32.89 <0.05 High-dose medicinal wine group 10 0.2 ml / 10 g 48±15 36.84 <0.05 Ankehaokuai group 10 0.15 ml / 10 g 34±12 42.11 <0.05

[0204] 2. Expectorant effect (phenol red method)

[0205] Healthy white mice, weighing 21±3 g, with an equal number of males and females, were randomly divided into 5 groups. Using ammonium chloride as the positive control, all groups were administered by gavage, and the experiment was conducted three days later. After 12 hours of fasting but with free access to water for each group of animals, 0.5 ml of 0.5 g / dl phenol red solution was injected intraperitoneally. Half an hour later, the animals were sacrificed by dislocation of the cervical vertebrae. The neck skin was cut open, and a 7-gauge injection needle without a tip was inserted into the trachea below the larynx, ligated and fixed. Using a 1-ml syringe, 0.5 ml of 5 g / dl NaHCO 3 solution was used to lavage the respiratory tract 3 times, and then the lavage fluid was aspirated, dropped into a colorimetric cuvette, and flushed 3 times in the same way. The aspirated washing fluids were combined and colorimetrically analyzed with a standard phenol red tube using a photoelectric colorimeter at the same time. Calculate the amount of phenol red in the aspirated washing fluid of each group each time. The more the content, the better the expectorant effect. The results are shown in Table 13.

[0206] Table 13 Effects of the Chinese medicinal wine prepared in Example 1 on the content of phenol red in the tracheal aspirate of mice X-±SD

[0207] Group Number of animals Dose Phenol red excretion (μg / ml) Relative percentage (%) P value Normal saline group 10 0.97±0.09 100 Xi'an Tequ group 10 0.15 ml / 10 g 0.94±0.12 100.00 >0.05 Low-dose medicinal wine group 10 0.1 ml / 10 g 1.53±0.40 157.73 <0.05 High-dose medicinal wine group 10 0.2 ml / 10 g 1.82±0.32 187.63 <0.01 Ammonium chloride group 10 14 mg / 10 g 1.73±0.43 178.35 <0.01

[0208] As can be seen from the results in Table 13, the amount of phenol red excreted from the tracheas of animals in the high-dose and low-dose groups of the Chinese medicinal wine prepared in Example 1 increased significantly, indicating that the Chinese medicinal wine prepared in Example 1 has an obvious expectorant effect.

[0209] VII. Anti-aging effect

[0210] 1. Effect on the tensile strength of rat tail tendon collagen fibers

[0211] Thirty healthy adult white rats, with an equal number of males and females, weighing 280±30 g, were randomly divided into 5 groups. After continuous gavage administration for 7 days according to the doses in Table 16, the experiment was conducted. Using total gypenosides as the positive control. After the animals were anesthetized with ether, their tails were cut off and placed in water at -10°C. At a position 1 - 1.5 cm from the tip of the tail, the skin was incised, and a bundle of collagen fibers was dissected. The collagen fibers were further divided into single fibers (Φ = 0.15 mm, length 3 - 4 cm) in cold physiological saline and fixed in a bath containing urea buffer solution (40°C) with a tension of 2 g. Record the time when the collagen fiber breaks. The shorter the time, the stronger the anti-aging effect. The results were compared with the saline group. The results are shown in Table 14.

[0212] Table 14 Effects of the Chinese medicinal wine prepared in Example 1 on the tensile strength of rat tail tendon collagen fibers X-±SD

[0213] Group Number of animals Dose Tensile strength (minutes) Shrinkage rate (%) P value Normal saline group 6 — 48.7±3.9 0.00 — Xi'an Tequ group 6 1.5 ml / 100 g 47.9±4.1 1.64 >0.05 Low-dose medicinal wine group 6 1.0 ml / 100 g 36.2±3.6 25.67 <0.05 High-dose medicinal wine group 6 2.0 ml / 100 g 30.4±2.3 37.58 <0.05 Gynostemma pentaphyllum group 6 1.4 mg / 100 g 31.3±2.7 35.72 <0.05

[0214] The results in Table 14 show that the tensile effect of the animal tail tendon collagen fibers in the traditional Chinese medicine medicated wine group prepared in Example 1 is significantly reduced, indicating that the traditional Chinese medicine medicated wine prepared in Example 1 can reduce the degree of fibrosis and hardness of the rat tail tendon, showing an obvious anti-aging effect.

[0215] 2. Influence on the immune system

[0216] 1) Immune cell antibody secretion function test (spectrophotometer method)

[0217] Fifty ICR strain white mice, half male and half female, with a body weight of 25±2 g, were randomly divided into 5 groups and given drugs by gavage. After continuous administration for seven days, each mouse was intraperitoneally injected with 0.2 ml of 5% sheep red blood cell suspension. Four days later, the animals were decapitated and sacrificed, and the spleens were taken and made into a spleen cell suspension of 15 mg spleen / ml with PBS. Then, 0.5 ml of the spleen cell suspension, 0.2% sheep red blood cells, and 1:10 guinea pig serum were added to the test tubes in sequence, incubated in a 37°C water bath for 1 hour, centrifuged at 300 rpm for 5 minutes, the supernatant was taken, and the optical density (OD value) was measured by colorimetry at 413 mm with a 722 grating spectrophotometer. Levamisole was used as the positive control, and the results are shown in Table 15.

[0218] Table 15 Influence of the traditional Chinese medicine medicated wine prepared in Example 1 on the cellular immune function of mice X-±SD

[0219] Group Number of animals Dose OD value Enhancement rate (%) P value Normal saline group 10 — 0.36 0.00 Xi'an Tequ group 10 0.15 ml / 10 g 0.35 0.00 >0.05 Low-dose medicinal wine group 10 0.1 ml / 10 g 0.48 33.33 <0.05 High-dose medicinal wine group 10 0.2 ml / 10 g 0.49 36.11 <0.05 Levamisole group 10 8 μg / 10 g 0.53 47.22 <0.01 。

[0220] The results in Table 15 show that the OD values of the animals in the high-dose and low-dose groups of the traditional Chinese medicine medicated wine prepared in Example 1 are significantly increased (P<0.05). This indicates that the antibody generation ability and antibody-forming cells of the immune cells of this group of animals are significantly improved, suggesting that the traditional Chinese medicine medicated wine prepared in Example 1 has the effect of enhancing the cellular immune ability of animals.

[0221] 2) E-rosette test

[0222] Fifty ICR strain white mice, with a body weight of 20±2 g, half male and half female, were randomly divided into 5 groups and given drugs by gavage for 3 days. One hour after the last administration, the animals were sacrificed by decapitation, 4 ml of the prepared lymphocyte suspension was taken, washed three times, the supernatant was discarded, an equal volume of 1% sheep red blood cell suspension was added, mixed well, centrifuged at 500 rpm for 5 minutes, fixed with 0.8% glutaraldehyde for 15 minutes, the supernatant was discarded, the precipitate was taken and dropped on a glass slide, spread into a film, dried and stained, and the positive rate of rosette formation in 200 lymphocytes was counted under the microscope. Levamisole was used as the positive control. The results are shown in Table 16.

[0223] Table 16 Influence of the traditional Chinese medicine medicated wine prepared in Example 1 on the E-rosette formation rate of mouse lymphocytes X-±SD

[0224] Group Number of animals Dose E rosette formation rate (%) P value Normal saline group 10 — 15.43±1.20 Xi'an Tequ group 10 0.15 ml / 10 g 15.03±1.80 >0.05 Low-dose medicinal wine group 10 0.1 ml / 10 g 20.85±2.79 <0.05 High-dose medicinal wine group 10 0.2 ml / 10 g 24.05±3.01 <0.05 Levamisole group 10 8.0 μg / 10 g 26.01±1.99 <0.05 。

[0225] The results shown in Table 16 above indicate that the Chinese medicinal wine prepared in Example 1 can significantly increase the E-rosette formation rate of mouse lymphocytes (P<0.05), indicating that it can enhance the humoral immunity.

[0226] The above experimental results show that the Chinese medicinal wine prepared in Example 1 of the present invention can significantly improve the fatigue resistance of mice in forced swimming and the cold tolerance at low temperature, and prolong the time of tolerance to hypoxia under normal pressure, indicating that the Chinese medicinal wine prepared in Example 1 has an obvious effect of improving and enhancing the physical fitness of animals.

[0227] The Chinese medicinal wine prepared in the present invention can significantly increase the pain threshold of mice induced by hot plate and reduce the number of writhing times of mice induced by chemicals, indicating that the Chinese medicinal wine prepared in Example 1 has a good analgesic effect.

[0228] It can significantly reduce the ear swelling degree of mice induced by croton oil and the paw swelling degree of rats induced by carrageenan, and alleviate the proliferative inflammation, indicating that the Chinese medicinal wine prepared in Example 1 has an obvious anti-inflammatory effect.

[0229] It can significantly increase the animal sleep rate of subthreshold dose of sodium pentobarbital; improve the correct rate of the safe area of the "Y"-type maze, and also improve the correct rate of old mice after training. It shows that the Chinese medicinal wine prepared in Example 1 can produce obvious effects of sedation and tranquilization, improving memory and memory retention ability.

[0230] It can increase the body weight of animals with spleen deficiency syndrome induced by rhubarb, raise the body temperature, and improve the general conditions such as activity and diet. It shows that the Chinese medicinal wine has the effects of strengthening the spleen and promoting appetite, and strengthening the body.

[0231] It can increase the Hb and RBC counts of rats with qi and blood deficiency syndrome induced by acetylphenylhydrazine, significantly reduce the WBC and Heinz body counts (P<0.05), and improve the mesenteric microcirculation of animals, indicating that the Chinese medicinal wine has the effects of tonifying qi and blood, and promoting blood circulation to remove blood stasis.

[0232] The Chinese medicinal wine significantly reduces the number of coughs of mice induced by ammonia water within 2 minutes, and significantly increases the phenol red content in the tracheal extract, indicating that the Chinese medicinal wine has an obvious antitussive and expectorant effect.

[0233] The Chinese medicinal wine significantly reduces the tensile strength of the tail tendon collagen fibers of rats, delays sclerosis, significantly increases the number of antibody-forming cells in the spleen of mice and their antibody-forming ability, and significantly increases the E-rosette formation rate of mouse lymphocytes. It shows that the Chinese medicinal wine has obvious anti-aging and immune enhancing effects on the body.

[0234] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A ginseng and tortoise tonic wine, characterized in that: It is made from the following raw materials: Angelica, Astragalus, Codonopsis, Poria, Cornus officinalis, Chuanxiong, Atractylodes, Schisandra, Tangerine peel, Lycium barbarum, Saposhnikovia divaricata, Notopterygium, Poria, Ophiopogon japonicus, Rehmannia root, Rehmannia root, Jujube, and Tortoise shell glue.

2. The ginseng and tortoise-reinforcing wine according to claim 1, characterized in that: It is made of the following raw materials in parts by weight: 15-45 parts of Chinese Angelica, 30-90 parts of Astragalus, 15-45 parts of Codonopsis, 18-54 parts of Poria, 18-54 parts of Cornus, 10-30 parts of Ligusticum chuanxiong, 30-90 parts of Atractylodes, 12-36 parts of Schisandra, 18-54 parts of Tangerine Peel, 15-45 parts of Lycium barbarum, 10-30 parts of Saposhnikovia, 10-30 parts of Notopterygium, 30-90 parts of Poria, 7.5-22.5 parts of Ophiopogon, 35-105 parts of Rehmannia root, 30-90 parts of Rehmannia root, 35-105 parts of Jujube, and 10-30 parts of Tortoise Shell Glue.

3. A ginseng and tortoise tonic wine according to claim 2, characterized in that: It is made of the following raw materials in parts by weight: 15-30 parts of Chinese Angelica, 30-60 parts of Astragalus, 15-30 parts of Codonopsis, 18-36 parts of Poria, 18-36 parts of Cornus, 10-20 parts of Chuanxiong, 30-60 parts of Atractylodes, 12-24 parts of Schisandra, 18-36 parts of Tangerine Peel, 15-30 parts of Lycium barbarum, 10-20 parts of Saposhnikovia, 10-20 parts of Notopterygium, 30-60 parts of Poria, 7.5-15 parts of Ophiopogon, 35-70 parts of Rehmannia root, 30-60 parts of Rehmannia root, 35-70 parts of Jujube, and 10-20 parts of Tortoise Shell Glue.

4. A ginseng and tortoise tonic wine, characterized in that: It is made of the following raw materials in parts by weight: 15 parts of Chinese Angelicae Sinensis, 31.25 parts of Astragalus, 15 parts of Codonopsis Pilosula, 18.75 parts of Poria, 18.75 parts of Cornus Officinalis, 11.25 parts of Ligusticum chuanxiong, 31.25 parts of Atractylodes Macrocephala, 12.5 parts of Schisandrae Chinensis, 18.75 parts of Tangerine Peel, 15 parts of Lycium Barbarum, 11.25 parts of Saposhnikovia Divaricata, 11.25 parts of Notopterygium Incisi, 31.25 parts of Poria, 7.5 parts of Ophiopogon japonicus, 37.5 parts of Radix Rehmanniae, 31.25 parts of Rehmanniae Preparata, 37.5 parts of Jujube, and 11.25 parts of Tortoise Shell Glue.

5. The method for preparing the ginseng and turtle-enhancing wine according to any one of claims 1 to 4, characterized in that: The following steps are involved: Take Chinese angelica, astragalus, codonopsis, tuckahoe, cornus officinalis, ligusticum chuanxiong, atractylodes, schisandra chinensis, tangerine peel, wolfberry, siler, notopterygium, poria, ophiopogon, raw rehmannia root, cooked rehmannia root, jujube, and tortoise shell glue by weight, grind them, add crystal sugar, then add white wine, soak in a sealed container, steam, cool, store in a cellar, filter, dilute with water, centrifuge, and filter to obtain the product.

6. The method for preparing the ginseng and turtle-enhancing wine according to claim 5, characterized in that: The following steps are involved: Take Chinese angelica, astragalus, codonopsis, poria, cornus officinalis, ligusticum chuanxiong, atractylodes, schisandra chinensis, tangerine peel, wolfberry, siler, notopterygium, poria, ophiopogon, rehmannia root, prepared rehmannia root, jujube, and tortoise shell glue by weight, grind them, add crystal sugar, and then add 5 to 50 times the amount of white wine as the medicinal materials, soak in a sealed container for 24 to 48 hours, steam for 2 to 3 hours, cool, put into a cellar for 10 to 21 days, filter, dilute with water, centrifuge, and filter.

7. The identification method of Shengui Guben wine according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) Identification of reference herbs of Angelica sinensis and Ligusticum chuanxiong (1.1) Take Shengui Guben wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, shake it with ether, extract it, combine the extracts, evaporate it to dryness, and dissolve the residue with ethyl acetate to prepare the test solution; (1.2) Take another reference drug, Angelica sinensis and Ligusticum chuanxiong, add ether to each, soak in cold water, shake from time to time, filter, evaporate the filtrate, and dissolve the residue in ethyl acetate to prepare the reference drug solution; (1.3) The test solution and the control medicinal material solution in step (1) are respectively aspirated and spotted on the same silica gel G thin layer plate, and petroleum ether-ethyl acetate is used as a developing solvent. The plate is developed, taken out, dried, and examined under a UV lamp at 365 nm. In the chromatogram of the test sample, a fluorescent spot of the same color is displayed at the corresponding position of the chromatogram of the control medicinal material; (2) Identification of Loganin from Cornus officinalis (2.1) Take ginseng and turtle wine, evaporate it to near dryness, add ethyl acetate to dissolve it, heat it under reflux, filter it, evaporate the filtrate to dryness, and add methanol to dissolve the residue to prepare the test solution; (2.2) Take another loganin reference substance and add methanol to prepare a reference substance solution; (2.3) The test solution of step (2.1) and the reference substance of step (2.2) are respectively aspirated and spotted on the same silica gel G thin layer plate, and ethyl acetate-acetone-formic acid-water is used as a developing solvent. The plate is developed, taken out, air-dried, sprayed with 10% sulfuric acid ethanol solution, and baked until the spots are clearly colored; in the chromatogram of the test sample, spots of the same color are displayed at the corresponding positions in the chromatogram of the reference substance; (3) Identification of Schisandrin A in Schisandra chinensis (3.1) Take the ginseng and turtle wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, extract it with ethyl acetate, combine the extracts, concentrate them on a water bath, and use them as the test solution; (3.2) Take another reference substance of Schisandrin A and add ethyl acetate to prepare a reference solution; (3.3) Pipette the test solution of step (3.1) and the reference substance of step (3.2), spot them respectively on the same silica gel GF254 thin layer plate, use petroleum ether-ethyl acetate-formic acid as the developing solvent, develop, take out, dry, and examine under ultraviolet light at 254nm. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the reference substance. (4) Identification of Chinese wolfberry reference medicinal materials (4.1) Take the ginseng and turtle wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, extract it with ethyl acetate, combine the extracts, concentrate them on a water bath, and use them as the test solution; (4.2) Take wolfberry fruit as a control medicinal material, add ethyl acetate to soak, filter, and concentrate the filtrate as the control medicinal material solution; (4.3) Take the test sample solution and the wolfberry reference medicinal material solution, spot them on the same silica gel G thin layer plate, use ethyl acetate-chloroform-formic acid as the developing solvent, develop, take out, dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the wolfberry reference medicinal material, a fluorescent spot of the same color appears; (5) Identification of hesperidin in tangerine peel (5.1) Take Shengui Guben wine, steam it until there is no alcohol taste, add water and transfer it to a separatory funnel in batches, extract it with water-saturated n-butanol, combine the extracts, evaporate to dryness, and add methanol to dissolve the residue as the test solution; (5.2) Take another hesperidin reference substance and add methanol to prepare a reference substance solution; (5.3) Take dried tangerine peel as a control medicinal material, add methanol to ultrasonicate, filter, and concentrate the filtrate to serve as the control medicinal material solution; (5.4) Pipette the test solution, reference substance and reference medicinal material solution, spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-water as the developing solvent, develop, take out, dry, then use the upper layer solution of toluene-ethyl acetate-formic acid-water as the developing solvent, develop, take out, dry, spray with aluminum chloride test solution, and examine under ultraviolet light at 365nm. In the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference medicinal material and the reference substance, a fluorescent spot of the same color is displayed; (6) Identification of oleanolic acid (6.1) Take the ginseng and turtle wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, shake it with ether, extract it, combine the extracts, evaporate it to dryness, and dissolve the residue with ethyl acetate to prepare the test solution; (6.2) Take oleanolic acid reference substance and add ethyl acetate to prepare reference substance solution; (6.3) Pipette the test solution and oleanolic acid reference solution and spot them on the same silica gel G thin layer plate respectively. Use toluene-ethyl acetate-glacial acetic acid as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and blow with hot air until the spots are clearly colored. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the reference.

8. The identification method of Shengui Guben wine according to claim 7, characterized in that: The following steps are involved: (1) Identification of reference herbs of Angelica sinensis and Ligusticum chuanxiong (1.1) Take Shengui Guben wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, and extract it 2 to 3 times with ether, 20 to 30 ml each time. Combine the extracts, evaporate them to dryness, and dissolve the residue with ethyl acetate to prepare the test solution; (1.2) Take another reference drug, Angelica sinensis and Ligusticum chuanxiong, add ether to each, soak in cold water, shake from time to time, filter, evaporate the filtrate, and dissolve the residue in ethyl acetate to prepare the reference drug solution; (1.3) The test solution and the control medicinal material solution of step (1) are respectively aspirated and spotted on the same silica gel G thin layer plate, and petroleum ether-ethyl acetate in a volume ratio of 20:1 is used as a developing solvent, developed, taken out, dried, and examined under a UV lamp at 365 nm. In the chromatogram of the test sample, a fluorescent spot of the same color is displayed at the corresponding position of the chromatogram of the control medicinal material; (2) Identification of Loganin from Cornus officinalis (2.1) Take ginseng and turtle wine, evaporate it to near dryness, add ethyl acetate to dissolve it, heat and reflux for 1 to 2 hours, filter it, evaporate the filtrate to dryness, and add methanol to dissolve the residue as the test solution; (2.2) Take another loganin reference substance and add methanol to make a solution containing 1-5 mg of the reference substance per 1 ml; (2.3) The test solution of step (2.1) and the reference substance of step (2.2) are respectively aspirated and spotted on the same silica gel G thin layer plate, and developed with ethyl acetate-acetone-formic acid-water in a volume ratio of 12:8:1:1, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and baked at 10-50° C. until the spots are clearly colored; in the chromatogram of the test sample, spots of the same color are displayed at the corresponding positions in the chromatogram of the reference substance; (3) Identification of schisandrin A and schisandrin B in Schisandra chinensis (3.1) Take the ginseng and turtle wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, extract it with ethyl acetate 2 to 3 times, 20 to 30 ml each time, combine the extracts, concentrate them on a water bath, and use them as the test solution; (3.2) Take schisandrin A and schisandrin B reference substances and add ethyl acetate to prepare a mixed reference substance solution containing 1-5 mg per 1 ml; (3.3) Pipette the test solution of step (3.1) and the reference substance of step (3.2), spot them respectively on the same silica gel GF254 thin layer plate, use petroleum ether-ethyl acetate-formic acid in a volume ratio of 15:5:1 as the developing solvent, develop, take out, dry, and examine under ultraviolet light at 254nm. In the chromatogram of the test sample, spots of the same color are displayed at the corresponding positions in the chromatogram of the reference substance. (4) Identification of Chinese wolfberry reference medicinal materials (4.1) Take the ginseng and turtle wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, extract it with ethyl acetate 2 to 3 times, 20 to 30 ml each time, combine the extracts, concentrate them on a water bath, and use them as the test solution; (4.2) Take wolfberry reference medicinal material, add ethyl acetate and soak for 1 to 2 hours, filter, and concentrate the filtrate as the reference medicinal material solution; (4.3) Take the test sample solution and the control medicinal material solution, spot them on the same silica gel G thin layer plate, use ethyl acetate-chloroform-formic acid in a volume ratio of 3:2:1 as the developing solvent, develop, take out, dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the wolfberry control medicinal material, a fluorescent spot of the same color appears; (5) Identification of hesperidin in tangerine peel (5.1) Take Shengui Guben wine, steam it until there is no alcohol taste, add water and transfer it to a separatory funnel in batches, extract it 1 to 3 times with water-saturated n-butanol, 10 ml each time, combine the extracts, evaporate to dryness, add methanol to dissolve the residue, and use it as the test solution; (5.2) Take another hesperidin reference substance and add methanol to prepare a reference substance solution; (5.3) Take dried tangerine peel as a control medicinal material, add methanol to ultrasonicate, filter, and concentrate the filtrate to serve as the control medicinal material solution; (5.4) Pipette the test solution, reference substance and reference medicinal material solution, spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-water with a volume ratio of 100:17:13 as the developing solvent, develop, take out, dry, then use the upper layer solution of toluene-ethyl acetate-formic acid-water with a volume ratio of 20:10:1:1 as the developing solvent, develop, take out, dry, spray with aluminum chloride test solution, and examine under ultraviolet light at 365nm. In the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference medicinal material and the reference substance, a fluorescent spot of the same color is displayed; (6) Identification of oleanolic acid (6.1) Take the ginseng and turtle wine, steam it until there is no alcohol taste, cool it, transfer it to a separatory funnel with water, and extract it 2 to 3 times with ether, 20 to 30 ml each time. Combine the extracts, evaporate them to dryness, and dissolve the residue with ethyl acetate to prepare the test solution; (6.2) Take the oleanolic acid reference substance and add ethyl acetate to prepare a reference substance solution containing 1 to 5 mg per 1 ml; (6.3) Pipette the test sample solution and oleanolic acid reference solution and spot them on the same silica gel G thin layer plate respectively. Use toluene-ethyl acetate-acetic acid in a volume ratio of 14:4:0.5 as the developing solvent. Develop the plate, take it out, dry it, spray it with 10% sulfuric acid ethanol solution, and blow it with hot air until the spots are clearly colored. In the chromatogram of the test sample, a spot of the same color appears at the corresponding position in the chromatogram of the reference.

9. The quality detection method of Shengui Guben wine according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) Preparation of reference solution Take reference substance solutions of astragaloside IV, hesperidin, loganin, ferulic acid, oleanolic acid, schisandrin A and schisandrin B respectively, add ethanol solvent to prepare mixed reference substance solution; (2) Take the ginseng and turtle wine, evaporate it to dryness, add ethanol to ultrasonically extract it, filter the extract, and filter it through a 0.22 μm filter membrane to use as the test solution; (3) Establishment of standard curve The mixed reference solution of step (1) was taken and diluted into 5 concentrations in sequence, and filtered through a 0.22 μm filter membrane respectively. 5 μL of the solution was taken and injected into HPLC for analysis, and the standard curve equation was drawn with the concentration as the horizontal axis and the peak area as the vertical axis; (4) Take 10 μL of the test solution in step (2) and inject it into HPLC for analysis to obtain a chromatogram. According to the retention time of the reference substance, substitute the peak area into the standard curve equation to calculate the content of each component.

10. The quality detection method of Shengui Guben Wine according to claim 9, characterized in that: The following steps are involved: The chromatographic conditions of step (3) and step (4) are Waters Sun Fire C18 chromatographic column, the specification of the chromatographic column is 250mm×4.6mm, 5μm; methanol is used as phase A, 0.1% phosphoric acid water is used as phase B, gradient elution, 0~6.8min, 40%-60%A; 6.8~15min, 60%-75%A; 15~26min, 75%-85%A; 26~34min, 85%-40%A; 34~50min, 40%A; 50~80min, 60%A; flow rate: 1mL / min, column temperature: 30℃, detection wavelength 245nm.

11. The quality detection method of Shengui Guben Wine according to claim 1, characterized in that: The standard curve equations for each active ingredient are as follows: 。 12. Use of the Shengui Guben wine according to any one of claims 1 to 4 in anti-inflammatory, analgesic, cough and expectorant.

13. Use of the ginseng and turtle tonic wine according to any one of claims 1 to 4 in anti-aging and improving immunity.

14. Use of the Shengui Guben wine according to any one of claims 1 to 4 in preventing Alzheimer's disease.

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