Detection method for a spleen-strengthening and hangover-relieving mixture

Abstract

The present invention belongs to the field of biomedicine, and particularly relates to a detection method for a spleen-strengthening and hangover-relieving mixture. Combining the clinical experience of famous veteran traditional Chinese medicine doctors, the present invention is modified and added on the basis of Gehua Jiedu Decoction, taking into account strengthening the spleen and protecting the liver, promoting digestion and stopping vomiting, clearing heat and resolving dampness, and the spleen-strengthening and hangover-relieving empirical formula is composed as follows: Flos Puerariae, Radix Puerariae, Flos Chrysanthemi, Scutellariae Baicalensis Tablets, Fructus Citri Sarcodactylis, Rhizoma Atractylodis Macrocephalae, Radix Paeoniae Alba, Poria, Caulis Bambusae in Taenia, Cortex Moutan, Fermented Medicated Leaven, Rhizoma Bletillae, Pericarpium Citri Reticulatae, Fructus Amomi, Radix Glycyrrhizae. The present invention has carried out thin-layer identification, characteristic spectrum analysis, and quantitative analysis of index components on this preparation, and has the remarkable advantages of simple operation and easy popularization and use, and can be applied to the product quality analysis of this mixture.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to a detection method for a spleen-strengthening and hangover-relieving mixture. Background Art

[0002] In recent years, with the improvement of people's living standards and the change of drinking habits, the incidence of alcohol poisoning has been increasing, and it has become the third largest public health problem in the world after cardiovascular and cerebrovascular diseases and tumors. Moderate drinking is beneficial to the body, but excessive drinking or long-term heavy drinking can cause acute or chronic alcohol poisoning. The liver is the main organ for ethanol metabolism, and the most common complication of alcohol poisoning is alcoholic liver disease. It can also lead to weakened brain inhibitory function, reduced discrimination, memory, and comprehension, or even disappearance. Visual impairment, drowsiness, and vomiting may occur. Chronic poisoning is manifested as cardiovascular disorders, malnutrition, intellectual decline, chronic gastritis, degeneration of the heart, liver, and kidneys, and even liver cirrhosis. After drinking, patients may experience symptoms such as weakness, thirst, bitter taste and bad breath in the mouth, nausea, dryness and anorexia, diarrhea, etc. Some patients may experience these symptoms for several days, seriously affecting the quality of life. Western medicine commonly uses naloxone hydrochloride, Xingnaojing injection, etc. to promote wakefulness, and proton pump inhibitors, vitamin B6, etc. to protect the stomach. However, the curative effect is general, and it is difficult to improve symptoms such as dizziness, weakness, thirst, bitter taste and bad breath in the mouth, nausea, dryness and anorexia in patients. Therefore, there is an urgent need for a traditional Chinese medicine compound composition that can play a role in protecting the liver and strengthening the spleen while relieving hangovers, so as to enable the human body to quickly recover health.

[0003] (1) Kudzu Root

[0004] Kudzu root is the dried root of Pueraria lobata (Willd.) Ohwi, a plant of the Leguminosae family. In the Chinese Pharmacopoeia 2020 edition, puerarin is used as the reference component for the content determination of kudzu root medicinal materials.

[0005] Kudzu root is a Chinese herbal medicine food rich in various active ingredients such as isoflavones, triterpenes and triterpene saponins, coumarins, alkaloids, and kudzu root polysaccharides. Kudzu root isoflavones are the main active ingredients, and kudzu root isoflavones contain more than 20 effective ingredients, such as puerarin, daidzin, 3'-hydroxypuerarin, daidzein, genistein, formononetin, iridin, etc. In recent years, the research on the components of kudzu root has been relatively active. Modern pharmacological research shows that its main active ingredient, isoflavones, has a special effect in maintaining the stability of the vascular system and is commonly used in the adjuvant treatment of cardiovascular and cerebrovascular diseases in clinical practice. In addition, based on its functions such as antioxidant damage, anti-cell apoptosis, and improvement of blood lipid and blood glucose metabolism, certain progress has also been made in the treatment of diseases such as osteoporosis, alcohol poisoning, and premenstrual syndrome.

[0006] (2) Chrysanthemum

[0007] Chrysanthemum is the dried capitulum of Chrysanthemum morifolium Ramat. of the Compositae family. In the 2020 edition of the Chinese Pharmacopoeia, chlorogenic acid is used as the reference substance for thin-layer chromatography identification, and chlorogenic acid, luteoloside, and 3,5-O-dicaffeoylquinic acid reference substances are used as the index components for content determination.

[0008] Chrysanthemum contains a variety of chemical components, mainly flavonoids, volatile oils, phenylpropanoids, terpenoids, amino acids, etc. Among them, flavonoids and phenylpropanoid compounds are the main pharmacodynamic components of chrysanthemum. Li Shizhen's Compendium of Materia Medica states: Chrysanthemum can benefit the five meridians, regulate the four limbs, and treat head wind and heat; modern pharmacological research shows that Hangbai chrysanthemum has the effects of dissipating heat, detoxifying, improving eyesight, and reducing blood pressure. Its extract can improve myocardial nutrition, remove reactive oxygen free radicals, strengthen the resistance of capillaries, and reduce the content of fat and cholesterol in the blood; it also has certain effects on curbing tumors, delaying aging, and enhancing human immunity.

[0009] (3) Scutellaria baicalensis Georgi Tablets

[0010] Scutellaria baicalensis Georgi is the dried root of Scutellaria baicalensis Georgi of the Lamiaceae family.

[0011] Scutellaria baicalensis Georgi has diverse chemical components, mainly including flavonoids, phenolic acids, amino acids, trace elements, etc. Among them, flavonoids account for a relatively large effective component. Currently, more than 40 flavonoid components with identified structures have been found, including baicalein, baicalin, 5,7,4-trihydroxy-8-methoxyflavone, oroxylin A-7-glucuronide, etc. In addition to its effects of acting as an antibiotic, antiviral, anticancer, analgesic, and antipyretic, Scutellaria baicalensis Georgi also helps to protect the body's liver, immune system, and nervous system.

[0012] (4) Fructus Citri Sarcodactylis

[0013] Fructus Citri Sarcodactylis is the dried fruit of Citrus medica L. var. sarcodactylis Swingle of the Rutaceae family.

[0014] The chemical components of Fructus Citri Sarcodactylis mainly contain volatile oils, coumarins, flavonoids, polysaccharides, inorganic elements, and amino acids and other various bioactive substances (such as limettin, limonin, dimethoxycoumarin, trihydroxydimethoxyflavone, daucosterol, β-sitosterol, p-hydroxycinnamic acid, palmitic acid, succinic acid, limettin dimer, a small amount of diosmin, and hesperidin, etc.). Currently, it has been found that Fructus Citri Sarcodactylis has various pharmacological activities such as anti-inflammatory, antitumor, blood sugar regulation, and lipid-lowering. Summary of the Invention

[0015] In order to solve the above-mentioned existing technical problems, the present application provides the following technical solutions:

[0016] The present invention provides a detection method for a spleen-strengthening and hangover-relieving mixture. The spleen-strengthening and hangover-relieving mixture includes kudzu root, flos puerariae, chrysanthemum, scutellaria baicalensis, fingered citron, atractylodes macrocephala, white peony root, poria cocos, bamboo shavings with ginger, cortex moutan, six-fermented medicated leaven, bletilla striata, dried tangerine peel, liquorice root, and amomum villosum. The detection method includes thin-layer identification of white peony root and fingered citron, and determination of the content of the target component baicalin in scutellaria baicalensis by liquid chromatography:

[0017] Among them, the identification method for white peony root and fingered citron includes the following steps:

[0018] S11: Take an appropriate amount of the spleen-strengthening and hangover-relieving mixture, add ethyl acetate for extraction, concentration, and re-dissolution to obtain a test solution A1;

[0019] S12: Weigh a white peony root control sample or a fingered citron control sample, and add ethyl acetate to prepare a control solution B1;

[0020] S13: Spot the test solution A1 and the control solution B1 on a stationary phase respectively and place them in a developing agent for development. Observe the results after drying;

[0021] The determination of the content of baicalin includes the following steps:

[0022] S21: Take an appropriate amount of the spleen-strengthening and hangover-relieving mixture, dilute it with methanol, and filter to obtain a test solution A2;

[0023] S22: Weigh a baicalin control sample and add methanol to prepare a control solution B2;

[0024] S23: Respectively draw the test solution A2 and the control solution B2 and inject them into a liquid chromatograph for determination. Establish a linear regression equation based on the concentration and peak area in the determination result of the control solution B2;

[0025] S24: Determine the content of baicalin in the test solution A2 according to the linear regression equation and the peak area in the determination result of the test solution A2.

[0026] Preferably, the preparation method of the spleen-strengthening and hangover-relieving mixture includes the following steps:

[0027] S31: By weight, decoct 30 parts of kudzu root, 30 parts of flos puerariae, 12 parts of chrysanthemum, 10 parts of scutellaria baicalensis, 10 parts of fingered citron, 12 parts of atractylodes macrocephala, 20 parts of white peony root, 20 parts of poria cocos, 6 parts of dried tangerine peel, 10 parts of bamboo shavings with ginger, 15 parts of cortex moutan, 10 parts of six-fermented medicated leaven, 3 parts of bletilla striata, and 6 parts of liquorice root twice with water, and filter to obtain filtrates of various medicinal herb pieces; among them, the method for the first decoction is: add 6 times the total mass of the solids of water, soak for 0.5 h, decoct for 0.5 h, and filter; the method for the second decoction is: add 3 times the total mass of the solids remaining after the first decoction filtration of water, and decoct for 0.5 h;

[0028] S32: Mix the filtrates of the medicinal herb slices, add 6 parts of fructus amomi, concentrate under reduced pressure, and then add steviol glycoside and water and mix to obtain the spleen-strengthening and hangover mixture.

[0029] Preferably, in step S32, after mixing, stir evenly, filter, seal, and sterilize.

[0030] Preferably, it further includes a method for analyzing the characteristic spectrum of the drug, specifically: analyzing the liquid-phase test sample of the spleen-strengthening and hangover mixture and the liquid-phase test sample of a single medicinal herb by liquid chromatography, and using the medicinal herb with characteristic index components and no interference with other components as the index drug in the spleen-strengthening and hangover mixture; the index drug is selected from pueraria root, white peony root, chrysanthemum, and scutellaria baicalensis;

[0031] The chromatographic conditions of the liquid chromatography method for determining the index drug are as follows:

[0032] The chromatographic column is an octadecyl-modified silica gel chromatographic column, the flow rate is 1 mL / min, and the injection volume is 20 μL;

[0033] The detector is a DAD detector, and the detection wavelength is 200 - 400 nm;

[0034] Mobile phase A is acetonitrile, mobile phase B is 0.1 wt% phosphoric acid aqueous solution, and the total volume of mobile phase A and mobile phase B is 100%;

[0035] Gradient conditions: from 0 to 5 min, the volume of mobile phase A is 3%, from 5 to 10 min, the volume of mobile phase A is 3% - 14%, from 10 to 15 min, the volume of mobile phase A is 14%, from 15 to 20 min, the volume of mobile phase A is 14% - 22%, from 20 to 25 min, the volume of mobile phase A is 22%, from 25 to 30 min, the volume of mobile phase A is 22% - 40%, from 30 to 35 min, the volume of mobile phase A is 40%, from 35 to 40 min, the volume of mobile phase A is 40% - 100%, from 40 to 45 min, the volume of mobile phase A is 100%; among them, the detection wavelength of pueraria root is 250 nm; the detection wavelength of white peony root is 230 nm; the detection wavelength of chrysanthemum is 348 nm; the detection wavelength of scutellaria baicalensis is 280 nm.

[0036] Preferably, in step S11, the test sample solution A1 is obtained by shaking and extracting the spleen-strengthening and hangover mixture with ethyl acetate, evaporating under reduced pressure, and then dissolving in ethyl acetate.

[0037] Furthermore, the number of times of shaking extraction is two.

[0038] Preferably, in step S13, when the reference solution B1 is a Paeonia lactiflora reference sample, the developing solvent is a mixed solution of chloroform, ethyl acetate, methanol, and formic acid; the mass ratio of chloroform, ethyl acetate, methanol, and formic acid is 40:5:10:0.2.

[0039] Further, in step S13, after drying, it is sprayed with a developer and heated at 105 °C until color development; the developer is an ethanol solution of 5 wt% vanillin sulfuric acid; comparing the chromatograms of the test solution A1 and the reference solution B1, if purple-blue spots appear at the same position, it indicates that the sample to be tested contains Paeonia lactiflora.

[0040] Preferably, in step S13, when the reference solution B1 is a Citrus medica var. sarcodactylis reference sample, the developing solvent is a mixed solution of cyclohexane and ethyl acetate; the mass ratio of cyclohexane and ethyl acetate is 3:1.

[0041] Further, in step S13, after drying, the result is observed with a 365 nm ultraviolet lamp; comparing the chromatograms of the test solution A1 and the reference solution B1, if fluorescent spots appear at the same position for the test sample and the corresponding reference sample, it indicates that the test sample contains Citrus medica var. sarcodactylis.

[0042] Preferably, in steps S22 and S23, a reference solution with a concentration of 20 - 1040 μg / mL is prepared with methanol and determined under the following chromatographic conditions:

[0043] The chromatographic column is an octadecyl-modified silica gel chromatographic column, the column temperature is 30 °C; the injection volume is 20 μL; the flow rate is 1.0 mL / min; the detector is a DAD detector, and the detection wavelength for baicalin is 280 nm;

[0044] Mobile phase A is an aqueous solution of 0.1 wt% phosphoric acid, and mobile phase B is acetonitrile; the total volume of mobile phase A and mobile phase B is 100%, and gradient elution is performed; the gradient elution conditions are as follows: the volume of mobile phase B is 3% from 0 - 5 min, 3% - 14% from 5 - 10 min, 14% from 10 - 15 min, 14 - 22% from 15 - 20 min, 22% from 20 - 25 min, 22 - 40% from 25 - 30 min, 40% from 30 - 35 min, 40 - 95% from 35 - 40 min, 95% from 40 - 45 min, 95% to 3% from 45 - 50 min, and 3% from 50 - 60 min.

[0045] Preferably, in the step S23, the linear regression equation is y = 18.806x + 62.539, R 2 = 0.9999; where y is the peak area, x is the concentration, and x = 26 - 416 μg·mL -1 .

[0046] The technical solution of the present invention has the following advantages compared with the prior art:

[0047] Combined with the clinical experience of famous veteran traditional Chinese medicine doctors, on the basis of Gegen Jiedu Decoction, it is modified and added, taking into account strengthening the spleen and protecting the liver, promoting digestion and stopping vomiting, clearing heat and resolving dampness, and forming the following empirical formula for strengthening the spleen and sobering up: Flos Puerariae, Radix Puerariae, Flos Chrysanthemi, Scutellariae Radix, Fructus Citri Sarcodactylis, Rhizoma Atractylodis Macrocephalae, Radix Paeoniae Alba, Poria, Caulis Bambusae In Taeniis, Cortex Moutan, Massa Medicata Fermentata, Bletillae Rhizoma, Pericarpium Citri Reticulatae, Fructus Amomi, Radix Glycyrrhizae. In the formula, Flos Puerariae is the monarch drug, which relieves alcohol and awakens the spleen; the minister drugs are Massa Medicata Fermentata for promoting digestion and regulating the stomach, Rhizoma Atractylodis Macrocephalae for strengthening the spleen and drying dampness, Radix Paeoniae Alba for soothing the liver and nourishing the liver, Fructus Amomi for promoting appetite and awakening the spleen, and Poria for promoting diuresis and percolating dampness; the assistant drug is Caulis Bambusae In Taeniis for resolving phlegm and stopping vomiting; Radix Puerariae for promoting the production of body fluid and quenching thirst, Flos Chrysanthemi, Scutellariae Radix, and Cortex Moutan for clearing heat and detoxifying, Bletillae Rhizoma for protecting the stomach and promoting granulation, Fructus Citri Sarcodactylis for regulating the stomach and relieving pain, and Pericarpium Citri Reticulatae for strengthening the spleen and resolving phlegm; Radix Glycyrrhizae for coordinating the actions of various drugs.

[0048] According to the prescription of the spleen-strengthening and sobering-up mixture, referring to the requirements of the Chinese Pharmacopoeia and relevant guidelines, thin-layer identification, inspection items such as relative density, pH value, and solid content, as well as characteristic spectrum analysis and quantitative analysis of index components were carried out on this preparation. During the development of the spleen-strengthening and sobering-up mixture, it was found that there was a lack of relevant detection methods for this preparation. Through systematic qualitative and quantitative analysis and research on this preparation, the present invention was completed. Description of the Drawings

[0049] Figure 1 It is the thin-layer identification diagram of Radix Paeoniae Alba; among them, 1 is the reference medicinal material of Radix Paeoniae Alba, 2 to 4 are the test samples, and 5 is the negative control of Radix Paeoniae Alba;

[0050] Figure 2 It is the thin-layer identification diagram of Fructus Citri Sarcodactylis; among them, 1 is the reference medicinal material of Fructus Citri Sarcodactylis, 2 to 4 are the test samples, and 5 is the negative control of Fructus Citri Sarcodactylis;

[0051] Figure 3 It is the characteristic spectrum of HPLC of the concentrated solution of the spleen-strengthening and sobering-up mixture; the spectra in the figure are the spectra at 230 nm, 250 nm, 280 nm, and 348 nm respectively;

[0052] Figure 4 It is the HPLC characteristic spectrum of Radix Paeoniae Alba in the spleen-strengthening and sobering-up mixture at 230 nm;

[0053] Figure 5 It is the HPLC characteristic spectrum of Radix Puerariae in the spleen-strengthening and sobering-up mixture at 250 nm;

[0054] Figure 6It is the HPLC characteristic spectrum of Flos Chrysanthemi in the Jianpi Xingjiu Mixture at 348 nm;

[0055] Figure 7 It is the HPLC characteristic spectrum of Radix Scutellariae in the Jianpi Xingjiu Mixture at 280 nm;

[0056] Figure 8 It is the HPLC characteristic spectrum of baicalin reference substance solution at 280 nm;

[0057] Figure 9 It is the HPLC characteristic spectrum of the test solution at 280 nm;

[0058] Figure 10 It is the HPLC characteristic spectrum of the negative sample solution of Radix Scutellariae at 280 nm. Specific Embodiments

[0059] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, so that those skilled in the art can better understand the present invention and be able to implement it, but the examples given are not intended to limit the present invention.

[0060] Traditional Chinese medicine decoction pieces: Pueraria lobata (batch number: 211117), Flos Puerariae lobatae (batch number: 220214), Flos Chrysanthemi (batch number: 220531), Radix Scutellariae tablets (batch number: 220607), Citrus medica sarcodactylis (batch number: 220125), Atractylodes macrocephala (batch number: 211010), Radix Paeoniae Alba (batch number: 220525), Poria cocos (batch number: 211231), Caulis Bambusae in Taeniis (batch number: 220506), Cortex Moutan (batch number: 220224), Shenqu (batch number: 220527), Bletilla striata (batch number: 220517), Pericarpium Citri Reticulatae (batch number: 220415), Licorice tablets (batch number: 220305), Amomum villosum (batch number: 220321), etc., 16 kinds of decoction pieces, all purchased from Nantong Sanyue Traditional Chinese Medicine Decoction Pieces Co., Ltd.

[0061] Control medicinal materials: Radix Paeoniae Alba control medicinal material, batch number 120905 - 202011; Citrus medica sarcodactylis control medicinal material, batch number 120933 - 201706; Radix Scutellariae (batch number: 120955 - 201810); Pueraria lobata (batch number: 121551 - 201805); all purchased from the National Institutes for Food and Drug Control.

[0062] Reagents: Thin layer chromatography silica gel G type, purchased from Qingdao Ocean Chemical Industry Co., Ltd.; analytical grade cyclohexane, chloroform, n - hexane, etc., Jiangsu Qiangsheng Functional Chemical Co., Ltd.; toluene, acetone, etc., Yonghua Chemical Co., Ltd.; Wahaha pure water, Hangzhou Wahaha Group Co., Ltd.

[0063] Example 1 Preparation of the mixture and negative sample

[0064] Preparation of the pilot-scale sample of the Spleen-Strengthening and Hangover-Relieving Mixture: Each batch number (batch numbers are JP20230101, JP20230102, and JP20230103) is prepared according to the preparation method, and 300 L is prepared for each batch number, with 500 mL per bottle.

[0065] The preparation method of the Spleen-Strengthening and Hangover-Relieving Mixture consists of the following steps:

[0066] (1) Prescription composition: 180 g of Pueraria lobata, 180 g of Flos Puerariae, 72 g of Chrysanthemum morifolium, 60 g of Scutellaria baicalensis slices, 60 g of Citrus medica var. sarcodactylis, 72 g of Atractylodes macrocephala, 120 g of Paeonia lactiflora, 120 g of Poria cocos, 36 g of Citrus reticulata Blanco, 60 g of Caulis Bambusae in Taeniis, 90 g of Cortex Moutan, 60 g of Six-Flavored Medicinal Leaven, 18 g of Bletilla striata, 36 g of Glycyrrhiza uralensis Fisch. slices, 36 g of Amomum villosum (added later), a total of 15 herbs.

[0067] (2) Preparation method: Weigh the medicinal herb slices according to the prescription amount in (1). Except for Amomum villosum, the other fourteen kinds of slices are decocted twice with water. For the first time, add 6 times the amount of water of the slices, soak for 0.5 h, decoct for 0.5 h, and filter; for the second time, add 3 times the amount of water of the slices, decoct for 0.5 h, and filter. Combine the above filtrates, add the Amomum villosum slices, concentrate under reduced pressure to about 900 mL, add an appropriate amount of steviol glycoside, adjust the total amount to 1000 mL with water, stir well, filter, seal, and sterilize to obtain the product.

[0068] Negative control test samples: 15 kinds of negative control test samples lacking single herbs. According to the prescription amount of the Spleen-Strengthening and Hangover-Relieving prescription, after removing the corresponding single herbs from the original prescription respectively, the preparation method is referred to that of the Spleen-Strengthening and Hangover-Relieving Mixture.

[0069] Single-herb test samples: Prepared respectively according to the methods under the thin-layer identification of each herb in the Chinese Pharmacopoeia 2020 edition.

[0070] Reference substance test samples: Solutions of puerarin, paeoniflorin, and baicalin reference substances are prepared respectively according to the methods under the thin-layer identification of each herb in the Chinese Pharmacopoeia 2020 edition.

[0071] Example 2

[0072] (1) Thin-layer identification of Paeonia lactiflora

[0073] Take 3 batches of samples and shake well (batch numbers are JP20230101, JP20230102, and JP20230103), measure 20 mL, extract twice with ethyl acetate by shaking, 20 mL each time, take the ethyl acetate layer for evaporation under reduced pressure, dissolve the residue in 2 mL of ethyl acetate to obtain the test solution; weigh 1.0 g of Paeonia lactiflora medicinal herb, add 2 mL of ethyl acetate, extract by shaking to obtain the Paeonia lactiflora reference herb solution; take 5 mL of the negative sample lacking Paeonia lactiflora, extract twice with ethyl acetate by shaking, 5 mL each time, and obtain the negative control solution lacking Paeonia lactiflora according to the preparation method of the test solution.

[0074] According to the test under the thin-layer chromatography method in Part IV of the Chinese Pharmacopoeia (2020 Edition), 10 μL, 10 μL, and 2.5 μL of the above three solutions were respectively taken and spotted on the same silica gel G thin-layer plate. Using chloroform-ethyl acetate-methanol-formic acid (40:5:10:0.2) as the developing agent, develop, take out, dry, spray with 5% vanillin sulfuric acid ethanol solution, and heat until the spots are clearly visible.

[0075] The results of thin-layer chromatography are shown in Figure 1 .

[0076] The three batches of test samples showed the same blue-violet spots at the corresponding positions as the Paeonia lactiflora Pall. reference medicinal material in the thin-layer chromatography. The spots were clear and distinct, with good resolution, and the Paeonia lactiflora Pall. negative control had no interference, so it was included in the detection method.

[0077] (2) Thin-layer identification of Citrus medica L. var. sarcodactylis Swingle

[0078] Respectively take 20 mL of the three batches of samples and shake well (batch numbers are JP20230101, JP20230102, JP20230103), extract twice with ethyl acetate by shaking, 20 mL each time, take the ethyl acetate layer and concentrate it under reduced pressure to dryness. Dissolve the residue in 2 mL of ethyl acetate to obtain the test solution; weigh 0.5 g of the Citrus medica L. var. sarcodactylis Swingle medicinal material, add 5 mL of ethyl acetate, shake and extract, and take the continuous filtrate as the Citrus medica L. var. sarcodactylis Swingle medicinal material solution; take 5 mL of the negative sample without Citrus medica L. var. sarcodactylis Swingle, extract twice with ethyl acetate by shaking, 5 mL each time, and obtain the negative control solution without Citrus medica L. var. sarcodactylis Swingle according to the preparation method of the test solution. According to the experiment under the thin-layer chromatography method in Part IV of the Chinese Pharmacopoeia (2020 Edition), 10 μL, 10 μL, and 2.5 μL of the above three solutions were respectively taken and spotted on the same silica gel G thin-layer plate. Using cyclohexane-ethyl acetate (3:1) as the developing agent, develop, take out, dry in the air, and examine under an ultraviolet lamp (365 nm) to observe the color and position of the fluorescent spots of the test sample and the medicinal material.

[0079] The results of thin-layer chromatography are shown in Figure 2 . The thin-layer chromatography of the Citrus medica L. var. sarcodactylis Swingle reference medicinal material showed fluorescent spots mainly at R f 0.3; the three batches of test samples showed the same main spots as the Citrus medica L. var. sarcodactylis Swingle reference medicinal material in the thin-layer chromatography position, with good resolution, and the Citrus medica L. var. sarcodactylis Swingle negative control had no interference, so it was included in the detection method.

[0080] Example 3

[0081] Three batches of pilot-scale samples JP20230101, JP20230102, and JP20230103 were used for inspection according to the relevant regulations in Part IV of the Chinese Pharmacopoeia (2020 Edition).

[0082] (1) Relative density

[0083] According to the relative density determination method in General Chapter 0601 of the Fourth Part of the Chinese Pharmacopoeia (2020 Edition), the relative density of three batches of pilot-scale samples was determined (at room temperature), and the results are shown in Table 1.

[0084] (2) pH value

[0085] According to the pH value determination method in General Chapter 0631 of the Fourth Part of the Chinese Pharmacopoeia (2020 Edition), the pH values of three parallel samples were determined using pH test paper, and the results are shown in Table 1.

[0086] (3) Others

[0087] This product is a preparation with multi-dose packaging. It was inspected according to the "volumetric method" under the [Minimum Fill Inspection Method] (General Chapter 0942 of the Fourth Part of the Pharmacopoeia of the People's Republic of China, 2020 Edition), and the fill volume should comply with the regulations. The results are shown in Table 1.

[0088] (4) Total solids

[0089] Refer to the test method for the determination of total solids in the Chinese Pharmacopoeia (2020 Edition) (General Chapter 3101). The Jianpi Xingjiu Mixture was pre-shaken, and 1 mL of the Jianpi Xingjiu Mixture was precisely measured with a pipette and placed in a 50 mL evaporating dish (previously weighed to constant weight), and dried in an oven at 105 °C to constant weight (the difference in weight between two consecutive weighings does not exceed 5 mg), and then precisely weighed. Calculate according to the following formula:

[0090]

[0091] In the formula, c x is the total solids of the test sample, g / mL -1 ; W is the weight of the test sample after weighing to constant weight, g; V is the volume of the test sample, mL.

[0092] Reference quality analysis of the concentrated solution of the Jianpi Xingjiu Mixture: The three samples used were observed. Compared with its decoction, the color of the concentrated solution was slightly darker, and there were slightly precipitates that were easily dispersed by gentle shaking; it had an aromatic smell and a pungent, slightly sour, and bitter taste. Due to the influence of process parameters such as volume change during the preparation process of the concentrated solution compared with the decoction, the content fluctuated greatly, but it did not affect the stability of the finished product content. Inspected according to the general chapter of the Chinese Pharmacopoeia, the average relative density of the concentrated solution of the Jianpi Xingjiu Mixture was 1.06 (at room temperature), the pH value was 4.6, and the solid content was 10.58%.

[0093] Table 1 Quality analysis of the Jianpi Xingjiu Mixture

[0094] Serial number Relative density pH Solid content (%) Filling volume JP20230101 1.06 4.6 10.72 Not less than 500 mL JP20230102 1.04 4.6 10.37 Not less than 500 mL JP20230103 1.06 4.6 10.64 Not less than 500 mL Average value 1.05 4.6 10.58 Not less than 500 mL

[0095] (5) Microbiological limit inspection

[0096] The suitability test of the microbial limit inspection method was carried out according to the 2020 Edition of Chinese Pharmacopoeia, Volume IV, 1105 and 1106, and a suitable inspection method was established.

[0097] For the three batches of pilot-scale samples, according to the results of the suitability test: the number of aerobic bacteria, molds and yeasts was counted by the plate method; the control bacterium Escherichia coli was detected by the conventional method, and the results are shown in Table 2 below.

[0098] Table 2 Results of the microbial limit inspection of three batches of samples

[0099]

[0100] According to the microbial limit inspection method in the 2020 Edition of Chinese Pharmacopoeia, Volume IV, 1105 and 1106, the total number of aerobic bacteria per milliliter of Jianpi Xingjiu Mixture should be < 10 2 cfu / mL, the total number of molds and yeasts should be < 10 cfu / mL, and Escherichia coli (1 mL) should not be detected. The three batches of pilot-scale samples of Jianpi Xingjiu Mixture all meet the standards.

[0101] Example 4

[0102] Using Jianpi Xingjiu Mixture and related single herbs, negative control herbs, etc. as samples, relevant liquid chromatography conditions were studied and established to analyze the main chemical composition and the source of single herbs in this preparation, providing reference for the present invention.

[0103] (1) Raw materials and reagents

[0104] Sliced herbs, reference herbs, negative control test samples: the same as in Example 1.

[0105] (2) Preparation of test samples

[0106] Preparation of the test sample solution of Jianpi Xingjiu Mixture: Accurately measure 5 mL of the Jianpi Xingjiu Mixture solution, place it in a 25 mL volumetric flask, add methanol and shake well, make up to the scale line, let stand, take an appropriate amount of the solution, and filter it through a 0.22 μm organic microporous membrane to obtain the test sample solution.

[0107] Preparation of the test sample solution of single herbs: Prepared respectively according to the methods under the thin-layer identification of each herb in the 2020 Edition of Chinese Pharmacopoeia.

[0108] ① Paeonia lactiflora Pall.: Accurately weigh 1.019 g of this product, place it in a stoppered conical flask, accurately add 100 mL of 30% ethanol, weigh, ultrasonically treat for 45 min, cool to room temperature, weigh again, and make up the lost weight with 60% ethanol, shake well, and let stand. Take an appropriate amount of the solution and filter it through a 0.22 μm organic microporous membrane to obtain the test sample solution.

[0109] ② Kudzu Root: Weigh accurately 1.026 g of this product, place it in a stoppered conical flask, accurately add 100 ml of 30% ethanol, weigh, ultrasonically treat for 45 min, cool to room temperature, weigh again, make up the lost weight with 30% ethanol, shake well, and let stand. Take an appropriate amount of the solution and filter it through a 0.22 μm organic microporous membrane to obtain the test solution.

[0110] ③ Scutellaria Root: Weigh accurately 1.001 g of this product, place it in a stoppered conical flask, accurately add 100 ml of 70% ethanol, weigh, ultrasonically treat for 45 min, cool to room temperature, weigh again, make up the lost weight with 70% ethanol, shake well, and let stand. Take an appropriate amount of the solution and filter it through a 0.22 μm organic microporous membrane to obtain the test solution.

[0111] ④ Chrysanthemum: Weigh accurately 1.01 g of this product, place it in a stoppered conical flask, accurately add 100 ml of 70% methanol, tightly stopper, weigh, ultrasonically treat for 45 min, cool, weigh again, make up the lost weight with 70% methanol, shake well, and let stand. Take an appropriate amount of the solution and filter it through a 0.22 μm organic microporous membrane to obtain the test solution.

[0112] (3) Chromatographic conditions

[0113] Agilent 1260 Infinity Ⅱ (DAD detector) system, Agilent Technologies, Inc., USA. The chromatographic column is Cosmosil 5C18-MS-Ⅱ (4.6 mm × 250 mm, 5 μm); using acetonitrile (A)-0.1% phosphoric acid aqueous solution (B) as the mobile phase for gradient elution (0 - 5 min, 3% A; 5 - 10 min, 3% - 14% A; 10 - 15 min, 14% A; 15 - 20 min, 14% - 22% A; 20 - 25 min, 22% A; 25 - 30 min, 22% - 40% A; 30 - 35 min, 40% A; 35 - 40 min, 40% - 100% A; 40 - 45 min, 100% A); the detection wavelength is the full wavelength of 200 - 400 nm; the flow rate is 1.0 mL·min -1 ; the injection volume is 20 μL.

[0114] (4) Results

[0115] The characteristic spectrum detection results of the Spleen-Strengthening and Hangover Mixture are as Figure 3 . Under the above optimized chromatographic conditions, there are 4 main characteristic peaks in this mixture. After comparison with the chromatograms of 4 medicinal materials ( Figures 4 to 7)By comparing separately and analyzing in combination with the Chinese Pharmacopoeia and relevant literature, it was confirmed that chromatographic peak 1 (puerarin) was from Pueraria lobata, peak 2 (paeoniflorin) was from Paeonia lactiflora Pall., peak 3 was from Chrysanthemum morifolium Ramat., and peak 4 (baicalin) was from Scutellaria baicalensis Georgi. Among other medicinal flavors, the main chromatographic peaks of each medicinal material could not be detected yet possibly due to their characteristic components being liposoluble components or the corresponding low contents of the medicinal materials in the mixture.

[0116] In the spleen-strengthening and hangover-relieving mixture, the main components that could be identified were from Pueraria lobata, Paeonia lactiflora Pall., Chrysanthemum morifolium Ramat., and Scutellaria baicalensis Georgi, which was consistent with the results of thin-layer identification. The main components could be selected for subsequent content analysis. In addition, the components and resolution in the characteristic spectrum of this preparation were poor.

[0117] Example 5

[0118] According to the above results of thin-layer identification and characteristic analysis, the content of the main component baicalin in this preparation was determined.

[0119] (1) Raw materials, reagents and methods

[0120] Ⅰ Experimental instruments, raw materials and reagents

[0121] Experimental instruments: Agilent 1260 Infinity Ⅱ (DAD detector): Agilent Technologies, Inc., USA; FA1004 electronic balance: Shanghai Shunyu Hengping Scientific Instrument Co., Ltd.

[0122] Baicalin (batch number 110715 - 202223, content 97.2%): Shanghai Yuanye Bio-Technology Co., Ltd.; spleen-strengthening and hangover-relieving mixture, negative sample without Scutellaria baicalensis Georgi: Hai'an City Traditional Chinese Medicine Hospital; Scutellaria baicalensis Georgi (batch number: 33 - 1 - 05): Nantong Sanyue Chinese Herbal Pieces Co., Ltd.; HPLC-grade methanol, acetonitrile: OCEANPAK, Sweden; chromatographic-grade phosphoric acid: Shanghai Aladdin Biochemical Technology Co., Ltd.; Wahaha purified water: Hangzhou Wahaha Group Co., Ltd.

[0123] Ⅱ Preparation of test samples

[0124] Preparation of test solution of spleen-strengthening and hangover-relieving mixture: Precisely measure 2 mL of spleen-strengthening and hangover-relieving mixture solution, place it in a 25 mL volumetric flask, add methanol, shake well, make up the volume to the scale, let it stand, take an appropriate amount of the solution and filter it through a 0.22 μm organic microporous filter membrane, and take the subsequent filtrate, that is obtained. Prepare the negative sample without Scutellaria baicalensis Georgi in the same way.

[0125] Preparation of reference solution: Precisely weigh 10.40 mg of baicalin reference substance, place it in a 10 mL volumetric flask, add an appropriate amount of methanol and dissolve it by ultrasonic wave. After cooling to room temperature, make up the volume to obtain the reference substance stock solution, in which the mass concentration of baicalin is 1040 μg·mL -1 .

[0126] Reference crude drug: Weigh 1.001 g of Scutellaria baicalensis Georgi accurately, place it in a conical flask with a stopper, accurately add 100 ml of 70% ethanol, weigh it, ultrasonically treat it for 45 minutes, cool it to room temperature, weigh it again, make up the lost weight with 70% ethanol, shake well, and let it stand. Take an appropriate amount of the solution, filter it through a 0.22 μm organic microporous membrane to obtain the reference crude drug solution.

[0127] Ⅲ Chromatographic conditions

[0128] The chromatographic column is Cosmosil 5C 18 -MS-Ⅱ (250 mm × 4.6 mm, 5 μm); with acetonitrile (B)-0.1% phosphoric acid aqueous solution (A) as the mobile phase for gradient elution (0 - 5 min, 3% B; 5 - 10 min, 3% - 14% B; 10 - 15 min, 14% B; 15 - 20 min, 14% - 22% B; 20 - 25 min, 22% B; 25 - 30 min, 22% - 40% B; 30 - 35 min, 40% B; 35 - 40 min, 40% - 95% B; 40 - 45 min, 95% B; 45 - 50 min, 95% - 3% B; 50 - 60 min, 3% B); the detection wavelength of baicalin is 280 nm; the flow rate is 1.0 mL·min -1 ; the column temperature is 30 °C; the injection volume is 20 μL.

[0129] (2) Methodological investigation

[0130] Ⅰ Specificity result investigation

[0131] Precisely pipette 20 μL each of the reference substance solution and the test solution, inject the samples according to the above chromatographic conditions, and investigate the specificity of this method.

[0132] Results: The results of the specificity investigation are shown in Figures 8 to 10 , compared with the reference substance solution and the reference crude drug, the chromatogram of the test solution shows chromatographic peaks with the same retention time at the same detection wavelength, and the negative sample solution without Scutellaria baicalensis Georgi has no corresponding chromatographic peak. It shows that the negative has no interference and the specificity of this method is good.

[0133] Ⅱ Linear relationship investigation

[0134] Precisely pipette 0.25, 0.5, 1, 2, 3, 4 mL of the reference substance stock solution respectively, place them in 10 mL volumetric flasks, add methanol to the scale, shake well, filter through a 0.22 μm organic microporous membrane, take the filtrate, and prepare baicalin mass concentrations of 26.00 μg·mL -1 , 52.00 μg·mL -1 , 104.0 μg·mL -1 , 208.0 μg·mL -1 , 312.0 μg·mL -1, a series of reference substance solutions with a concentration of 416.0 μg·mL -1 , and each reference substance has 6 concentrations. Inject and analyze according to the above chromatographic conditions respectively. Using the mass concentration (μg·mL -1 ) of the reference substance as the abscissa (x) and the integrated peak area value as the ordinate (y), calculate the linear regression equation and range.

[0135] Result: The regression equation of baicalin is y = 18.806x + 62.539, R 2 = 0.9999. The linear relationship of baicalin concentration is good in the range of 26.00 - 416.0 μg·mL -1 .

[0136] Ⅲ Precision investigation

[0137] Take the reference substance solution with a baicalin concentration of 208.0 μg·mL -1 under the linear relationship investigation, accurately pipette 20 μL, inject samples 6 times under the above chromatographic conditions, and measure successively. Calculate the relative standard deviation (RSD) of the peak area of the baicalin chromatographic peak to investigate the precision.

[0138] The results of the precision investigation are shown in Table 3. The results of the precision investigation show that the RSD of the peak area of baicalin is 0.13%, and the instrument precision is good.

[0139] Table 3 Results of the precision investigation for the determination of baicalin content in Jianpi Xingjiu Mixture

[0140] Number 1 2 3 4 5 6 RSD% Peak area 3823 3818 3828 3830 3832 3825 0.13

[0141] Ⅳ Repeatability investigation

[0142] Take 6 portions of the same batch of Jianpi Xingjiu Mixture, prepare the test solution according to the above method for preparing the test solution, accurately pipette 20 μL, inject samples under the above chromatographic conditions, and measure. Calculate the RSD value of the peak area of the baicalin chromatographic peak to investigate the repeatability.

[0143] The results of the repeatability investigation are shown in Table 4. The results of the repeatability investigation show that the RSD of the peak area of baicalin is 0.45%, and the reproducibility is good, which is applicable to this method for preparing the test solution.

[0144] Table 4 Results of the repeatability investigation for the determination of baicalin content in Jianpi Xingjiu Mixture

[0145] Number 1 2 3 4 5 6 RSD% Peak area 2881 2914 2898 2910 2887 2891 0.45

[0146] Ⅴ Stability investigation

[0147] Precisely pipette 2 mL of the Jianpi Xingjiu Mixture, prepare the test solution according to the preparation method of the test solution, precisely pipette 20 μL of the test solution, and inject and measure at 0, 2, 4, 8, 12, and 24 h respectively according to the above chromatographic conditions. Calculate the RSD value of the peak area of the baicalin chromatographic peak respectively to investigate the stability.

[0148] The results of the stability investigation are shown in Table 5. The investigation results show that the RSD of the baicalin peak area is 0.82%, and the test solution has good stability within 24 h.

[0149] Table 5 Results of the stability investigation for the determination of baicalin content in the Jianpi Xingjiu Mixture

[0150] Time (h) 1 2 3 4 5 6 Average value RSD% Peak area 2781 2774 2792 2783 2816 2834 2797 0.82

[0151] Ⅵ Spike recovery

[0152] Precisely pipette 0.5 mL of the Jianpi Xingjiu Mixture with known baicalin content into a 5 mL volumetric flask, precisely add 1 mL of the reference substance solution of baicalin (150.2 μg·mL -1 ) dissolved in methanol, treat according to the preparation method of the test solution, prepare 6 portions in parallel, take an appropriate amount of the solution and filter it through a 0.22 μm organic microporous filter membrane, and take the filtrate to obtain the test solution. Measure the peak area of the baicalin chromatographic peak in the test solution according to the above chromatographic conditions, calculate the concentration of the baicalin component in the test solution, and then calculate the spike recovery (spike recovery (%) = (measured value of the spiked sample - measured value of the sample) / spiked amount × 100%).

[0153] The results of the spike recovery are shown in Table 6. The average spike recovery of baicalin is %, between 95.00% and 105.00%, and the RSD value is 0.34%, indicating that this method can be used for the determination of baicalin content in the Jianpi Xingjiu Mixture.

[0154] Table 6 Results of the spike recovery investigation for the determination of baicalin content in the Jianpi Xingjiu Mixture

[0155]

[0156] (3) Baicalin content in the Jianpi Xingjiu Mixture

[0157] Prepare the test solutions of three batches of the Jianpi Xingjiu Mixture according to the preparation method of the test solution under item (1) of this example. Inject and measure according to the chromatographic conditions under item (1) of this example. Each of the two parallel samples in each batch is injected once, and the results are averaged. The results of the content determination of the three batches of the Jianpi Xingjiu Mixture are shown in Table 7. The baicalin content of the three batches of mixtures is in the range of 1.231 to 1.286 mg·mL -1 range.

[0158] Table 7 Results of the determination of baicalin content in three batches of the Jianpi Xingjiu Mixture

[0159]

[0160] (4) Summary

[0161] The present invention has established a method for determining baicalin in the spleen-strengthening and hangover mixture. The determination results of three batches of test samples show that the contents of the two index components are stable, and the detection method is feasible.

[0162] During the research process of the present invention, it was found that the content of baicalin in the test samples is stable and can be selected as the index component for the investigation of the detection method.

[0163] Obviously, the above embodiments are merely examples given for clear illustration and are not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to enumerate all the implementation manners here. And the obvious changes or modifications derived therefrom are still within the protection scope of the present invention.

Claims

1. A method for detecting a spleen-strengthening and alcohol-relief mixture, characterized in that: The detection method includes thin layer identification of white peony root and bergamot root, and liquid chromatography to determine the content of baicalin, an indicator component in scutellaria baicalensis root; Wherein, the method for identifying white peony root and bergamot root comprises the following steps: S11: Take an appropriate amount of the spleen-strengthening and hangover-revitalizing mixture, add ethyl acetate to extract, concentrate and redissolve to obtain a test solution A1; S12: Weigh a white peony root control sample or a citron root control sample, and add ethyl acetate to prepare a reference solution B1; S13: Spot the test solution A1 and the reference solution B1 on the stationary phase respectively and place them in a developing agent for development, and observe the results after drying; The determination of the content of baicalin comprises the following steps: S21: Take an appropriate amount of the spleen-strengthening and hangover-relief mixture, add methanol to dilute, and filter to obtain the test solution A2; S22: weighing a baicalin reference sample and adding methanol to prepare a reference solution B2; S23: respectively aspirate the test solution A2 and the reference solution B2 and inject them into the liquid chromatograph for determination, and establish a linear regression equation according to the concentration and peak area of ​​the determination result of the reference solution B2; S24: Determine the content of baicalin in the test solution A2 according to the linear regression equation and the peak area in the test result of the test solution A2; in the steps S22 and S23, the determination is performed under the following chromatographic conditions: the chromatographic column is a carbon 18 modified silica gel chromatographic column, the column temperature is 30°C; the injection volume is 20 μL; the flow rate is 1.0 mL / min; the detector is a DAD detector, and the detection wavelength for determining baicalin is 280 nm; The preparation method of the spleen-strengthening and hangover-relief mixture comprises the following steps: S31: By weight, 30 parts of kudzu root, 30 parts of kudzu flower, 12 parts of chrysanthemum, 10 parts of scutellaria, 10 parts of bergamot, 12 parts of atractylodes, 20 parts of white peony root, 20 parts of tuckahoe, 6 parts of dried orange peel, 10 parts of ginger bamboo, 15 parts of peony bark, 10 parts of jiaoshenliuqu, 3 parts of bletilla striata and 6 parts of liquorice are respectively decocted twice with water, and filtered to obtain filtrates of various medicinal materials; wherein the first decoction method is: adding 6 times the total weight of solid water, soaking for 0.5 h, decocting for 0.5 h, and filtering; the second decoction method is: adding 3 times the total weight of solid water remaining after filtering in the first decoction, and decocting for 0.5 h; S32: mixing the filtrate of the medicinal material slices, adding 6 parts of Amomum villosum, concentrating under reduced pressure, adding stevioside and water to mix, and obtaining the spleen-strengthening and hangover-relief mixture; When the chromatography is used for determination, the mobile phase A is a 0.1wt% phosphoric acid aqueous solution, and the mobile phase B is acetonitrile; the total volume of the mobile phase A and the mobile phase B is 100%, and gradient elution is performed; the gradient elution conditions are: the volume of the mobile phase B is 3% at 0-5 min, the volume of the mobile phase B is 3%-14% at 5-10 min, the volume of the mobile phase B is 14% at 10-15 min, the volume of the mobile phase B is 14% at 15-20 min, the volume of the mobile phase B is 14-22% at 20-25 min, the volume of the mobile phase B is 22% at 25-30 min, the volume of the mobile phase B is 22-40% at 30-35 min, the volume of the mobile phase B is 40% at 30-40 min, the volume of the mobile phase B is 40-95% at 35-40 min, the volume of the mobile phase B is 95% at 40-45 min, the volume of the mobile phase B is 95% at 45-50 min, and the volume of the mobile phase B is 95% to 3% at 50-60 min.

2. The detection method of the spleen-strengthening and alcohol-relief mixture as claimed in claim 1, characterized in that: The invention also includes a characteristic spectrum analysis method of the drug, specifically: analyzing the liquid phase test sample of the spleen-strengthening and alcohol-revitalizing mixture and the liquid phase test sample of a single medicinal material by liquid chromatography, and using the medicinal material having characteristic index components and not interfering with other components as the index drug in the spleen-strengthening and alcohol-revitalizing mixture; the index drugs are kudzu root, white peony root, chrysanthemum and scutellaria; The chromatographic conditions of the liquid chromatography method for determining the index drug are as follows: The chromatographic column was a carbon 18-modified silica gel column, the flow rate was 1 mL / min, and the injection volume was 20 μL; The detector is a DAD detector with a detection wavelength of 200-400nm; Mobile phase A was acetonitrile, mobile phase B was 0.1 wt% phosphoric acid aqueous solution, and the total volume of mobile phase A and mobile phase B was 100%; Gradient conditions: 3% volume of mobile phase A at 0-5 min, 3%-14% volume of mobile phase A at 5-10 min, 14% volume of mobile phase A at 10-15 min, 14% volume of mobile phase A at 15-20 min, 14%-22% volume of mobile phase A at 20-25 min, 22% volume of mobile phase A at 25-30 min, 22%-40% volume of mobile phase A at 30-35 min, 40% volume of mobile phase A at 35-40 min, 40%-100% volume of mobile phase A at 40-45 min, and 100% volume of mobile phase A at 40-45 min; the detection wavelength of Pueraria lobata was 250 nm; the detection wavelength of white peony root was 230 nm; the detection wavelength of chrysanthemum was 348 nm; and the detection wavelength of Scutellaria baicalensis was 280 nm.

3. The detection method of the spleen-strengthening and alcohol-relief mixture as claimed in claim 1, characterized in that: In the step S11, the test solution A1 is obtained by extracting the spleen-strengthening and hangover-relief mixture twice by shaking with ethyl acetate, evaporating under reduced pressure, and then adding ethyl acetate to dissolve it.

4. The detection method of the spleen-strengthening and alcohol-relief mixture as claimed in claim 1, characterized in that: In step S13, when the reference solution B1 is a white peony root reference sample, the developing agent is a mixed solution of chloroform, ethyl acetate, methanol and formic acid; wherein the mass ratio of chloroform, ethyl acetate, methanol and formic acid is 40:5:10:0.

2.

5. The detection method of the spleen-strengthening and alcohol-relief mixture as claimed in claim 4, characterized in that: In the step S13, after drying, spray with a color developer and heat at 105°C until color is developed; the color developer is a 5 wt% vanillin sulfuric acid ethanol solution; the chromatograms of the test solution A1 and the reference solution B1 are compared, and if blue-purple spots appear at the same position, it means that the sample to be tested contains white peony root.

6. The detection method of the spleen-strengthening and alcohol-relief mixture as claimed in claim 1, characterized in that: In step S13, when the reference solution B1 is a bergamot reference sample, the developing agent is a mixed solution of cyclohexane and ethyl acetate, wherein the mass ratio of cyclohexane to ethyl acetate is 3:

1.

7. The detection method of the spleen-strengthening and alcohol-relief mixture as claimed in claim 6, characterized in that: In the step S13, after drying, the results are observed using a 365nm ultraviolet lamp; the chromatograms of the test solution A1 and the reference solution B1 are compared, and if fluorescent spots appear at the same position in the test sample and the corresponding reference solution, it means that the test sample contains bergamot.

8. The detection method of the spleen-strengthening and alcohol-relief mixture as claimed in claim 1, characterized in that: In the steps S22 and S23, methanol is used to prepare a reference solution with a concentration of 20-1040 μg / mL.

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