Quality detection method of traditional Chinese medicine preparation for treating leukorrheal diseases

Abstract

The invention relates to a quality detection method of a traditional Chinese medicine preparation for treating leukorrheal diseases, and establishes a thin-layer chromatography identification method for soil-fried bighead atractylodes rhizome, fried Chinese yam, wine-fried white peony root, wine-fried semen plantaginis, bran-fried rhizoma atractylodis, ginseng, liquorice, radix bupleuri, pericarpium citri reticulatae and charred schizonepeta spike in the traditional Chinese medicine preparation for treating leukorrheal diseases. Meanwhile, the fingerprint spectrum of the traditional Chinese medicine preparation for treating leukorrheal diseases is established by adopting high performance liquid chromatography. According to the quality detection method of the traditional Chinese medicine preparation for treating the leukorrheal diseases, comprehensive quality evaluation can be carried out on the traditional Chinese medicine preparation for treating the leukorrheal diseases, the method is simple, convenient, high in accuracy and good in reproducibility, a scientific basis is provided for quality detection and evaluation, and the product quality is effectively controlled.

Description

Technical Field

[0001] The present application relates to the technical field of drug detection, and in particular to a quality detection method for a traditional Chinese medicine preparation for treating leucorrhea. Background Art

[0002] The name "leucorrhea" was first seen in Huangdi Neijing, while the name "leucorrhea disease" was first seen in Zhubingyuanhoulun. Leucorrhea can be divided into broad and narrow senses. The broad sense of leucorrhea refers to gynecological and obstetric diseases. Because these diseases occur below the belt meridian, they are called "leucorrhea". Narrow sense of leucorrhea includes physiological leucorrhea and pathological leucorrhea. Among them, pathological leucorrhea refers to abnormalities in the amount, color, quality, and taste of leucorrhea, or accompanied by systemic or local symptoms, so it is called "leucorrhea disease". In Western medicine, it is common to see an increase or decrease in leucorrhea caused by diseases such as vaginitis, cervicitis, pelvic inflammatory disease, premature ovarian failure, amenorrhea, infertility, and gynecological tumors. The main cause of leucorrhea disease is dampness, so it is difficult to cure, recurring, and often complicated by irregular menstruation, amenorrhea, infertility and other diseases. It is a common disease among female patients second only to menstrual diseases. Since ancient times, Chinese medicine has a unique advantage in the prevention and treatment of leucorrhea, and has been concerned by doctors of all generations. Therefore, providing a quality inspection method for traditional Chinese medicine preparations for treating leukorrhea and analyzing the experience of using medicines for treating leukorrhea can provide useful reference for the clinical treatment of leukorrhea.

[0003] Wandai Decoction is one of the 100 classic prescriptions in the "Catalogue of Ancient Classic Prescriptions (First Batch)" issued by the State Administration of Traditional Chinese Medicine. Wandai Decoction comes from "Fu Qingzhu's Gynecology". Its main ingredients include Atractylodes macrocephala, Chinese yam, ginseng, white peony, plantain seed, Atractylodes lancea, licorice (roasted), dried tangerine peel, black mustard spike, bupleurum, etc. These ingredients work together to achieve the effects of tonifying the spleen and soothing the liver, removing dampness and stopping leukorrhea. Wandai Decoction is suitable for the treatment of spleen deficiency, liver depression, dampness and turbidity leukorrhea, manifested by symptoms such as white leukorrhea, clear and thin like mucus, pale complexion, fatigue and loose stools, pale tongue with white fur, slow or weak pulse, etc. In order to enable the ancient classic prescriptions to be widely used in clinical practice and ensure their safety and effectiveness, it is very important to establish qualitative and quantitative methods for their quality evaluation. Summary of the invention

[0004] Based on this, it is necessary to provide a quality inspection method for traditional Chinese medicine preparations for the treatment of leucorrhea, which can conduct a comprehensive quality evaluation of traditional Chinese medicine preparations for the treatment of leucorrhea. The method is simple, accurate and reproducible, which can provide a scientific basis for quality inspection and evaluation and effectively control the quality of the product.

[0005] The technical solution of this application is as follows:

[0006] In one aspect of the present invention, a quality detection method for a Chinese medicine preparation for treating leukorrhea is provided, comprising constructing a fingerprint of the Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography, and identifying components of the Chinese medicine preparation for treating leukorrhea by thin layer chromatography; the raw materials of the Chinese medicine preparation for treating leukorrhea include the following components: stir-fried atractylodes macrocephala, stir-fried yam, white peony root with wine, plantain seed with wine, stir-fried atractylodes lancea with bran, ginseng, liquorice, bupleurum, dried orange peel and schizonepeta spicate charcoal;

[0007] The method of constructing the fingerprint of the traditional Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography comprises the following steps:

[0008] The Chinese herbal medicine for treating leukorrhea is extracted using an extraction solvent to prepare a test solution;

[0009] The test solution is subjected to high performance liquid chromatography detection to construct a fingerprint of the traditional Chinese medicine preparation for treating leucorrhea;

[0010] The method of identifying the components of the Chinese medicine preparation for treating leukorrhea by thin layer chromatography comprises the following steps:

[0011] Take a reference substance, add a dissolving solvent to dissolve it, and prepare a reference substance solution; the reference substance includes at least one of atractylodes lactone I, atractylodes lactone II, atractylodes lactone III, paeoniflorin, gallic acid, ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, liquiritin, isoliquiritigenin, ammonium glycyrrhizinate, saikosaponin a, saikosaponin d, and hesperidin;

[0012] The traditional Chinese medicine preparation for treating leucorrhea is extracted with water and an organic solvent, and the organic phases are combined to prepare a test solution;

[0013] Mix water and reference medicinal materials, decoct, filter and collect the filtrate to prepare a reference medicinal material solution; the reference medicinal materials include at least one of stir-fried Atractylodes macrocephala, stir-fried Chinese yam, stir-fried white peony root, stir-fried Plantago asiatica, stir-fried Atractylodes lancea with bran, ginseng, liquorice, bupleurum, dried orange peel and schizonepeta spicate charcoal;

[0014] The test solution, control medicinal material solution and / or control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a developing agent, dried, developed with a color developing agent, and inspected.

[0015] In one embodiment, the conditions of the high performance liquid chromatography method include: mobile phase A is aqueous formic acid solution, mobile phase B is acetonitrile, and gradient elution is used;

[0016] The procedure of the gradient elution includes:

[0017] 0-15min, the volume percentage of the mobile phase B is maintained at 5%;

[0018] 15min-60min, the volume percentage of the mobile phase B increases from 5% to 30%;

[0019] 60min-75min, the volume percentage of the mobile phase B increases from 30% to 85%;

[0020] 75min-85min, the volume percentage of the mobile phase B increases from 85% to 95%;

[0021] From 85 min to 90 min, the volume percentage of the mobile phase B is maintained at 95%.

[0022] In one embodiment, when constructing the fingerprint of the traditional Chinese medicine preparation for treating leucorrhea by high performance liquid chromatography, at least one of the following conditions is met:

[0023] (1) Flow rate: 0.8 mL / min-1.2 mL / min;

[0024] (2) Column temperature is 25°C-35°C;

[0025] (3) The detection wavelength is 200nm-400nm;

[0026] (4) The injection volume is 5 μL-20 μL;

[0027] (5) In the formic acid aqueous solution, the volume percentage of formic acid is 0.01%-0.15%;

[0028] (6) The extraction solvent is a methanol-water solution; optionally, the volume percentage of methanol in the methanol-water solution is 50%-90%.

[0029] In one embodiment, the conditions for identifying the Chinese medicine preparation for treating leucorrhea by thin layer chromatography include: the dissolving agent is at least one of methanol and ethanol; and / or

[0030] The organic phase is selected from at least one of n-butanol, ethyl acetate, dichloromethane and diethyl ether; and / or

[0031] The developing agent is at least one selected from the group consisting of cyclohexane, a mixture of ethyl acetate and toluene, a mixture of toluene and acetone, a mixture of ethyl acetate, methanol, formic acid and water, a mixture of chloroform, ethyl acetate, methanol and water, a mixture of ethyl acetate, formic acid, glacial acetic acid and water, a mixture of ethyl acetate, methanol and water, and a mixture of cyclohexane, ethyl acetate, formic acid and water; and / or

[0032] The color developer is selected from at least one of sulfuric acid ethanol solution, vanillin sulfuric acid solution, p-dimethylaminobenzaldehyde sulfuric acid solution and aluminum chloride ethanol solution.

[0033] In one embodiment, the method for thin layer chromatography identification of Atractylodes macrocephala and Atractylodes lancea in the traditional Chinese medicine preparation for treating leucorrhea comprises the following steps:

[0034] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0035] The control medicinal material solutions were prepared by mixing water, Atractylodes macrocephala control medicinal material and Atractylodes lancea control medicinal material, decocting, filtering and collecting the filtrate;

[0036] The reference substances of atractylodes lactone I, atractylodes lactone II and atractylodes lactone III were dissolved in methanol to prepare reference substance solutions;

[0037] The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, developed with a mixture of cyclohexane, ethyl acetate and toluene as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0038] In the developing solvent, the volume ratio of cyclohexane, ethyl acetate and toluene is (14-16):3:3;

[0039] The color developer is a sulfuric acid ethanol solution; optionally, the volume percentage of sulfuric acid in the sulfuric acid ethanol solution is 10%;

[0040] The method for thin layer chromatography identification of yam in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0041] Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and dichloromethane, combining the organic phases, and preparing a test solution;

[0042] Mix water and yam control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution;

[0043] The test solution and the control medicinal material solution are spotted on the same silica gel G thin layer plate respectively, and developed with a mixed solution of toluene and acetone as a developing agent, the silica gel G thin layer plate is dried, and a color developer is used for color development;

[0044] In the developing agent, the volume ratio of toluene to acetone is (4-6):1;

[0045] The color developer is a vanillin sulfuric acid solution; optionally, in the vanillin sulfuric acid solution, the volume percentage of vanillin is 5%.

[0046] In one embodiment, the method for thin layer chromatography identification of white peony root in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0047] Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and ethyl acetate, combining the organic phases, and preparing a test solution;

[0048] Mix water and white peony root control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution;

[0049] The reference substances of paeoniflorin and gallic acid were dissolved in ethanol to prepare reference substance solutions;

[0050] The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a mixture of ethyl acetate, methanol, formic acid and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 254 nm;

[0051] In the developing solvent, the volume ratio of ethyl acetate, methanol, formic acid and water is (15-17):2:1.5:1;

[0052] The color developer is a vanillin sulfuric acid solution; optionally, the volume percentage of vanillin in the vanillin sulfuric acid solution is 5%;

[0053] The method for thin layer chromatography identification of Plantago in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0054] Using water and ether, and water and n-butanol to extract the traditional Chinese medicine preparation for treating leucorrhea in sequence, combining the organic phases, and preparing a test solution;

[0055] Mix water and Plantago seed control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution;

[0056] The test solution and the control medicinal material solution are spotted on the same silica gel G thin layer plate, developed with a mixture of butyl acetate, methanol, formic acid and water as a developing agent, dried, and developed with a color developer;

[0057] In the developing agent, the volume ratio of butyl acetate, methanol, formic acid and water is (5-7): 1.5: 1.5: 1;

[0058] The color developer is a vanillin sulfuric acid solution; optionally, in the vanillin sulfuric acid solution, the volume percentage of vanillin is 0.5%.

[0059] In one embodiment, the method for thin layer chromatography identification of ginseng in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0060] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0061] The control medicinal material solution is prepared by mixing water and ginseng control medicinal material respectively, decocting, filtering and taking the filtrate;

[0062] The reference substances of ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1 were dissolved in methanol to prepare reference substance solutions;

[0063] The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate, developed with a mixture of chloroform, ethyl acetate, methanol and water as a developing agent, dried, developed with a color developer, and examined under sunlight and ultraviolet light at 365 nm, respectively;

[0064] In the developing solvent, the volume ratio of chloroform, ethyl acetate, methanol and water is (14-16): (39-41): 22: 10;

[0065] The color developer is a sulfuric acid ethanol solution; optionally, the volume percentage of sulfuric acid in the sulfuric acid ethanol solution is 10%;

[0066] The method for thin layer chromatography identification of liquorice in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0067] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0068] The control medicinal material solution is prepared by mixing water and ginseng control medicinal material respectively, decocting, filtering and taking the filtrate;

[0069] The reference substances, liquiritin, isoliquiritin and ammonium glycyrrhizinate, were dissolved in methanol to prepare reference substance solutions;

[0070] The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a mixed solution of ethyl acetate, formic acid, glacial acetic acid and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0071] In the developing solvent, the volume ratio of ethyl acetate, formic acid, glacial acetic acid and water is (14-16): 1: 1: 2;

[0072] The color developer is a sulfuric acid ethanol solution; optionally, in the sulfuric acid ethanol solution, the volume percentage of sulfuric acid is 10%.

[0073] In one embodiment, the method for thin layer chromatography identification of the Chinese medicinal preparation Bupleurum for treating leucorrhea comprises the following steps:

[0074] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0075] The control medicinal material solution was prepared by mixing water and the control medicinal material Bupleurum respectively, decocting, filtering and taking the filtrate;

[0076] Dissolve saikosaponin a and saikosaponin d reference substances in methanol to prepare reference substance solutions;

[0077] The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, developed with a mixture of ethyl acetate, methanol and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0078] In the developing solvent, the volume ratio of ethyl acetate, methanol and water is (7-9):2:1;

[0079] The color developer is a sulfuric acid solution of p-dimethylaminobenzaldehyde; optionally, the volume percentage of p-dimethylaminobenzaldehyde in the sulfuric acid solution of p-dimethylaminobenzaldehyde is 2%;

[0080] The method for thin layer chromatography identification of dried orange peel and schizonepeta spicate in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0081] Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and ethyl acetate, combining the organic phases, and preparing a test solution;

[0082] The control medicinal materials of dried tangerine peel and schizonepeta spicate were respectively mixed with water, decocted, filtered and the filtrate was collected to prepare a control medicinal material solution;

[0083] The hesperidin reference substance was dissolved in methanol to prepare a reference substance solution;

[0084] The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, first developed with a mixture of ethyl acetate, methanol and water for 4.5 cm, then developed with a mixture of cyclohexane, ethyl acetate, formic acid and water for 8 cm, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0085] In the developing solvent, the volume ratio of ethyl acetate, methanol and water is 100:(16-18):(12-14); the volume ratio of cyclohexane, ethyl acetate, formic acid and water is (9-11):(9-11):2:5;

[0086] The color developer is an aluminum chloride ethanol solution; optionally, in the aluminum chloride ethanol solution, the volume percentage of aluminum chloride is 5%.

[0087] In one embodiment, the raw materials of the Chinese medicine preparation for treating leucorrhea include the following components in parts by weight:

[0088] 20 parts of stir-fried Atractylodes macrocephala, 20 parts of stir-fried Chinese yam, 10 parts of white peony root with wine, 6 parts of plantain seeds with wine, 6 parts of stir-fried Atractylodes macrocephala with bran, 4 parts of ginseng, 2 parts of liquorice, 1.2 parts of bupleurum, 1 part of dried tangerine peel and 1 part of schizonepeta spicate charcoal;

[0089] Optionally, the Chinese medicine preparation for treating leucorrhea is one of the pills, powders, pastes, tablets, granules, lozenges, capsules, aqueous solutions or tinctures of Wandai Decoction.

[0090] Another aspect of the present invention provides the application of the quality detection method of the traditional Chinese medicine preparation for treating leucorrhea as described above in the quality control of the traditional Chinese medicine preparation for treating leucorrhea.

[0091] Compared with the prior art, this application has the following beneficial effects:

[0092] The present application provides a quality detection method for Chinese medicine preparations for treating leukorrhea, and establishes a thin layer chromatography identification method of stir-fried Atractylodes macrocephala, stir-fried Chinese yam, white peony root with wine, plantain seed with wine, stir-fried Atractylodes macrocephala with bran, ginseng, licorice, bupleurum, tangerine peel and schizonepeta spicate charcoal in Chinese medicine preparations for treating leukorrhea, and at the same time, a fingerprint of Chinese medicine preparations for treating leukorrhea is established by high performance liquid chromatography. Through the above-mentioned quality detection method for Chinese medicine preparations for treating leukorrhea, a comprehensive quality evaluation of Chinese medicine preparations for treating leukorrhea can be carried out, and the method is simple, accurate and reproducible, providing a scientific basis for quality detection and evaluation, and effectively controlling the quality of products. BRIEF DESCRIPTION OF THE DRAWINGS

[0093] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0094] Figure 1 The HPLC chromatograms of different decoction times of Wandai Decoction in Example 1 are shown.

[0095] Figure 2 The HPLC chromatograms of Wandai decoction before and after concentration in Example 1 are shown.

[0096] Figure 3 The HPLC chromatograms of Wandai decoction before and after freeze-drying in Example 1 are shown.

[0097] Figure 4 HPLC chromatograms of water and different concentrations of ethanol extracted in Example 1, from top to bottom: H 2 O, 50% ethanol, 70% ethanol and 90% ethanol extraction.

[0098] Figure 5 The HPLC chromatograms after extraction with different concentrations of methanol in Example 1 are shown.

[0099] Figure 6 HPLC chromatograms of different wavelengths in Example 1.

[0100] Figure 7 The optimized HPLC chromatogram in Example 1 is shown in FIG.

[0101] Figure 8 This is the chromatographic peak attribution diagram of the stir-fried Atractylodes macrocephala in Wandai Decoction in Example 1.

[0102] Fig. 9 This is the chromatographic peak attribution diagram of fried yam in Wandai soup in Example 1.

[0103] Fig.10 This is the attribution diagram of the chromatographic peaks of Plantago asiatica in Wandai Decoction of Example 1.

[0104] Fig.11 This is the chromatographic peak attribution diagram of the white peony root in Wandai Decoction in Example 1.

[0105] Fig.12 This is the chromatographic peak attribution diagram of the stir-fried Atractylodes macrocephala with bran in the Wandai soup of Example 1.

[0106] Fig.13 This is the chromatographic peak attribution diagram of ginseng in Wandai Decoction of Example 1.

[0107] Fig.14 This is the attribution diagram of the chromatographic peaks of liquorice in Wandai Decoction in Example 1.

[0108] Fig.15 This is the chromatographic peak attribution diagram of Radix Bupleuri in Wandai Decoction of Example 1.

[0109] Fig.16 This is the attribution diagram of the chromatographic peaks of dried orange peel in Wandai Decoction of Example 1.

[0110] Fig.17 This is the chromatographic peak attribution diagram of Schizonepeta spicata charcoal in Wandai decoction in Example 1.

[0111] Fig.18 It is the chromatogram of Wandai Decoction-gallic acid in Example 1.

[0112] Fig.19 This is the chromatogram of Wandai Decoction-peonyside in Example 1.

[0113] Fig. 20 This is the chromatogram of Wandai Decoction-Verbascoside in Example 1.

[0114] Fig.21 This is the chromatogram of Wandai Decoction-Hesperidin in Example 1.

[0115] Fig. 22It is the chromatogram of Wandai Decoction-ammonium glycyrrhizinate in Example 1.

[0116] Fig.23 This is the chromatogram of Wandai Decoction-Atractylodes lactone III in Example 1.

[0117] Fig.24 This is the chromatogram of Wandai Decoction-Glycyrrhizin in Example 1.

[0118] Fig.25 This is a summary diagram of the attribution of the chromatographic peaks of each medicinal material in Example 1.

[0119] Fig.26 This is a summary of the chromatographic peaks identified in Example 1.

[0120] Fig. 27 The chromatogram is a thin layer identification chromatogram of Wandai Decoction-Atractylodes macrocephala and Atractylodes lancea in Example 1. Among them, 1 is Atractylodes macrocephala and Atractylodes macrocephala double negative; 2 is Wandai Decoction; 3 is Atractylodes macrocephala; 4 is Atractylodes macrocephala; 5 is Atractylodes lactones I, II, and III (from top to bottom).

[0121] Fig.28 This is the thin layer identification chromatogram of Wandai Soup-Chinese Yam in Example 1, where A. is the immediate picture after color development; B. is the picture left overnight after color development.

[0122] Fig.29 This is the thin layer identification chromatogram of Wandai Decoction-White Peony Root in Example 1, wherein 1 is gallic acid + paeoniflorin, 2 is Wandai Decoction, 3 is White Peony Root with Wine, and 4 is White Peony Root with Wine negative.

[0123] Fig.30 This is the thin layer identification chromatogram of Wandai Decoction-Plantago Seed in Example 1, wherein 1 is Plantago Seed negative, 2 is Wandai Decoction, and 3 is Plantago Seed.

[0124] Fig.31 This is the thin layer identification chromatogram of Wandai Decoction-Ginseng in Example 1, wherein 1 is ginsenoside Rg1; 2 is ginsenoside Rb1; 3 is ginsenoside Re; 4 is ginseng negative; 5 is ginseng; 6 and 7 are Wandai Decoction.

[0125] Fig.32 This is the thin layer identification chromatogram of Wandai Decoction-Glycyrrhiza in Example 1, wherein 1 is the control of liquiritigenin, isoliquiritigenin, and ammonium glycyrrhizinate; 2 is Wandai Decoction; 3 is Glycyrrhiza; and 4 is the negative control of Glycyrrhiza.

[0126] Fig.33 It is the thin layer identification chromatogram (365nm) of Wandai Decoction-Bupleurum in Example 1, wherein 1 is saikosaponin a; 2 is Bupleurum negative; 3 is Bupleurum; 4 is Wandai Decoction 5μL; 5 is Wandai Decoction (ginseng); 6 is Wandai Decoction 10μL; 7 is saikosaponin d; 8 is saikosaponin a+saikosaponin d.

[0127] Fig.34 It is the thin layer chromatogram (365nm) of dried tangerine peel and schizonepeta spicate in Example 1, wherein 1 is hesperidin; 2 is dried tangerine peel negative; 3 is dried tangerine peel 2μL; 4 is dried tangerine peel 5μL; 5 is Wandai decoction; 6 is schizonepeta spicate charcoal 2μL; 7 is schizonepeta spicate charcoal 5μL. DETAILED DESCRIPTION

[0128] The present application will be further described in detail below in conjunction with specific embodiments. The present application can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present application more thoroughly understood.

[0129] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0130] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0131] As used herein, "one or more" refers to any one, any two, or any two or more of the listed items.

[0132] Herein, the optional scope of "and / or", "or / and", and "and / or" includes any one of two or more relevant listed items, and also includes any and all combinations of the relevant listed items, wherein said any and all combinations include any two relevant listed items, any more relevant listed items, or a combination of all relevant listed items.

[0133] Herein, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0134] Herein, "first aspect", "second aspect", "third aspect", "fourth aspect", etc. are used for descriptive purposes only and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. In addition, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present application, the meaning of "multiple" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined. In the description of the present application, the meaning of "several" is at least one, such as one, two, etc., unless otherwise clearly and specifically defined.

[0135] In this application, when it comes to numerical ranges, unless otherwise specified, the above numerical ranges are deemed to be continuous and include the minimum and maximum values ​​of the range, as well as each value between such minimum and maximum values. Further, when a range refers to an integer, each integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0136] The percentage contents involved in this application, unless otherwise specified, refer to mass percentage for solid-liquid mixing and solid-solid mixing, and refer to volume percentage for liquid-liquid mixing.

[0137] The percentage concentrations mentioned in this application, unless otherwise specified, refer to the final concentration, which refers to the percentage of the added component in the system after the addition of the component.

[0138] The temperature parameters in this application, unless otherwise specified, allow both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±5°C, ±2°C, ±1°C, ±0.5°C, ±0.4°C, ±0.3°C, ±0.2°C, and ±0.1°C are allowed. Normal temperature or room temperature in this application refers to no temperature control operation, generally 4°C to 35°C, preferably 20±5°C.

[0139] In one aspect of the present invention, a quality detection method for a Chinese medicine preparation for treating leukorrhea is provided, comprising constructing a fingerprint of the Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography, and identifying components of the Chinese medicine preparation for treating leukorrhea by thin layer chromatography; the raw materials of the Chinese medicine preparation for treating leukorrhea include the following components: stir-fried atractylodes macrocephala, stir-fried yam, white peony root with wine, plantain seed with wine, stir-fried atractylodes lancea with bran, ginseng, liquorice, bupleurum, dried orange peel and schizonepeta spicate charcoal;

[0140] The method of constructing the fingerprint of the traditional Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography comprises the following steps:

[0141] The Chinese herbal medicine for treating leukorrhea is extracted using an extraction solvent to prepare a test solution;

[0142] The test solution is subjected to high performance liquid chromatography detection to construct a fingerprint of the traditional Chinese medicine preparation for treating leucorrhea;

[0143] The method of identifying the components of the Chinese medicine preparation for treating leukorrhea by thin layer chromatography comprises the following steps:

[0144] Take a reference substance, add a dissolving solvent to dissolve it, and prepare a reference substance solution; the reference substance includes at least one of atractylodes lactone I, atractylodes lactone II, atractylodes lactone III, paeoniflorin, gallic acid, ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, liquiritin, isoliquiritigenin, ammonium glycyrrhizinate, saikosaponin a, saikosaponin d, and hesperidin;

[0145] The traditional Chinese medicine preparation for treating leucorrhea is extracted with water and an organic solvent, and the organic phases are combined to prepare a test solution;

[0146] Mix water and reference medicinal materials, decoct, filter and collect the filtrate to prepare a reference medicinal material solution; the reference medicinal materials include at least one of stir-fried Atractylodes macrocephala, stir-fried Chinese yam, stir-fried white peony root, stir-fried Plantago asiatica, stir-fried Atractylodes lancea with bran, ginseng, liquorice, bupleurum, dried orange peel and schizonepeta spicate charcoal;

[0147] The test solution, control medicinal material solution and / or control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a developing agent, dried, developed with a color developing agent, and inspected.

[0148] In some of the examples, the conditions of the HPLC method include: mobile phase A is formic acid aqueous solution, mobile phase B is acetonitrile, and gradient elution is used;

[0149] The procedure of the gradient elution includes:

[0150] 0-15min, the volume percentage of the mobile phase B is maintained at 5%;

[0151] 15min-60min, the volume percentage of the mobile phase B increases from 5% to 30%;

[0152] 60min-75min, the volume percentage of the mobile phase B increases from 30% to 85%;

[0153] 75min-85min, the volume percentage of the mobile phase B increases from 85% to 95%;

[0154] From 85 min to 90 min, the volume percentage of the mobile phase B is maintained at 95%.

[0155] In some examples, when constructing the fingerprint of the traditional Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography, at least one of the following conditions is met:

[0156] (1) Flow rate: 0.8 mL / min-1.2 mL / min;

[0157] (2) Column temperature is 25°C-35°C;

[0158] (3) The detection wavelength is 200nm-400nm;

[0159] (4) The injection volume is 5 μL-20 μL.

[0160] In some of the examples, the conditions of the high performance liquid chromatography also include: in the formic acid aqueous solution, the volume percentage of formic acid is 0.01%-0.15%.

[0161] In some examples, the conditions of the high performance liquid chromatography also include: the extraction solvent is a methanol-water solution.

[0162] In some examples, the volume percentage of methanol in the methanol aqueous solution is 50%-90%. Further, the volume percentage of methanol in the methanol aqueous solution is 50%-70%.

[0163] In some of the examples, constructing the fingerprint of the traditional Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography also includes the following steps:

[0164] Dissolving the reference substance to prepare a reference substance mixed solution;

[0165] Performing high performance liquid chromatography on the reference solution to obtain a reference spectrum;

[0166] According to the reference spectrum, the chromatographic peaks of the fingerprint spectrum of the traditional Chinese medicine preparation for treating leucorrhea are identified as common peaks;

[0167] Wherein, the reference substance includes one or more of gallic acid, paeoniflorin, verbascoside, hesperidin, ammonium glycyrrhizinate, liquiritin, atractylodes lactone I, atractylodes lactone II, atractylodes lactone III, bupleurosaponin a and glycyrrhizin.

[0168] In some of the examples, the chromatographic peaks of the fingerprint of the traditional Chinese medicine preparation for treating leucorrhea contain 14 chromatographic peaks, among which the identified characteristic peaks include: Peak 1 is earth-fried Atractylodes macrocephala, Peak 3 and Peak 7 are wine-fried white peony root, Peak 6 is fried yam and bran-fried Atractylodes macrocephala, Peak 8, Peak 9 and Peak 14 are licorice, Peak 10 is wine-fried Plantago seed, Peak 12 is wine-fried Plantago seed, and Peaks 12 and 13 are dried tangerine peel.

[0169] In some of the examples, the fingerprint of the traditional Chinese medicine preparation for treating leukorrhea was identified by comparing it with the retention time and ultraviolet absorption spectrum of the reference substance, and a total of 7 chromatographic peaks were identified, including: gallic acid, paeoniflorin, verbascoside, hesperidin, ammonium glycyrrhizinate, atractylodes lactone III, and glycyrrhizin.

[0170] In some of the examples, the conditions for identifying the traditional Chinese medicine preparation for treating leucorrhea by thin layer chromatography include: the dissolving reagent is at least one of methanol and ethanol.

[0171] In some of the examples, the conditions for identifying the traditional Chinese medicine preparation for treating leukorrhea by thin layer chromatography include: the organic phase is selected from at least one of n-butanol, ethyl acetate, dichloromethane and ether.

[0172] In some of the examples, the conditions for using thin layer chromatography to identify the traditional Chinese medicine preparation for treating leukorrhea include: the developing agent is selected from at least one of cyclohexane, a mixture of ethyl acetate and toluene, a mixture of toluene and acetone, a mixture of ethyl acetate, methanol, formic acid and water, chloroform, ethyl acetate, methanol and water, a mixture of ethyl acetate, formic acid, glacial acetic acid and water, a mixture of ethyl acetate, methanol and water, and a mixture of cyclohexane, ethyl acetate, formic acid and water.

[0173] In some of the examples, the conditions for using thin layer chromatography to identify the traditional Chinese medicine preparation for treating leukorrhea include: the color developer is selected from at least one of sulfuric acid ethanol solution, vanillin sulfuric acid solution, p-dimethylaminobenzaldehyde sulfuric acid solution and aluminum chloride ethanol solution.

[0174] In some of the examples, when the TLC method is used to identify the Chinese medicine preparation for treating leukorrhea, the inspection method includes: comparing the TLC of the test solution with the TLC of the reference solution to see whether there is an identical spot at a position corresponding to the TLC of the reference solution;

[0175] In some examples, the method for thin layer chromatography identification of Atractylodes macrocephala and Atractylodes lancea in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0176] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0177] The control medicinal material solutions were prepared by mixing water, Atractylodes macrocephala control medicinal material and Atractylodes lancea control medicinal material, decocting, filtering and collecting the filtrate;

[0178] The reference substances of atractylodes lactone I, atractylodes lactone II and atractylodes lactone III were dissolved in methanol to prepare reference substance solutions;

[0179] The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, developed with a mixture of cyclohexane, ethyl acetate and toluene as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0180] In the developing solvent, the volume ratio of cyclohexane, ethyl acetate and toluene is (14-16):3:3;

[0181] The color developer is a sulfuric acid ethanol solution; optionally, in the sulfuric acid ethanol solution, the volume percentage of sulfuric acid is 10%.

[0182] In some examples, the method for thin layer chromatography identification of yam in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0183] Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and dichloromethane, combining the organic phases, and preparing a test solution;

[0184] Mix water and yam control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution;

[0185] The test solution and the control medicinal material solution are spotted on the same silica gel G thin layer plate respectively, and developed with a mixed solution of toluene and acetone as a developing agent, the silica gel G thin layer plate is dried, and a color developer is used for color development;

[0186] In the developing agent, the volume ratio of toluene to acetone is (4-6):1;

[0187] The color developer is a vanillin sulfuric acid solution; optionally, in the vanillin sulfuric acid solution, the volume percentage of vanillin is 5%.

[0188] In some examples, the method for thin layer chromatography identification of white peony root in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0189] Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and ethyl acetate, combining the organic phases, and preparing a test solution;

[0190] Mix water and white peony root control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution;

[0191] The reference substances of paeoniflorin and gallic acid were dissolved in ethanol to prepare reference substance solutions;

[0192] The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a mixture of ethyl acetate, methanol, formic acid and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 254 nm;

[0193] In the developing solvent, the volume ratio of ethyl acetate, methanol, formic acid and water is (15-17):2:1.5:1;

[0194] The color developer is a vanillin sulfuric acid solution; optionally, in the vanillin sulfuric acid solution, the volume percentage of vanillin is 5%.

[0195] In some examples, the method for thin layer chromatography identification of Plantago in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0196] Using water and ether, and water and n-butanol to extract the traditional Chinese medicine preparation for treating leucorrhea in sequence, combining the organic phases, and preparing a test solution;

[0197] Mix water and Plantago seed control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution;

[0198] The test solution and the control medicinal material solution are spotted on the same silica gel G thin layer plate, developed with a mixture of butyl acetate, methanol, formic acid and water as a developing agent, dried, and developed with a color developer;

[0199] In the developing agent, the volume ratio of butyl acetate, methanol, formic acid and water is (5-7): 1.5: 1.5: 1;

[0200] The color developer is a vanillin sulfuric acid solution; optionally, in the vanillin sulfuric acid solution, the volume percentage of vanillin is 0.5%.

[0201] In some examples, the method for thin layer chromatography identification of ginseng in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0202] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0203] The control medicinal material solution is prepared by mixing water and ginseng control medicinal material respectively, decocting, filtering and taking the filtrate;

[0204] The reference substances of ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1 were dissolved in methanol to prepare reference substance solutions;

[0205] The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate, developed with a mixture of chloroform, ethyl acetate, methanol and water as a developing agent, dried, developed with a color developer, and examined under sunlight and ultraviolet light at 365 nm, respectively;

[0206] In the developing solvent, the volume ratio of chloroform, ethyl acetate, methanol and water is (14-16): (39-41): 22: 10;

[0207] The color developer is a sulfuric acid ethanol solution; optionally, in the sulfuric acid ethanol solution, the volume percentage of sulfuric acid is 10%.

[0208] In some examples, the method for thin layer chromatography identification of licorice in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0209] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0210] The control medicinal material solution is prepared by mixing water and ginseng control medicinal material respectively, decocting, filtering and taking the filtrate;

[0211] The reference substances, liquiritin, isoliquiritin and ammonium glycyrrhizinate, were dissolved in methanol to prepare reference substance solutions;

[0212] The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a mixed solution of ethyl acetate, formic acid, glacial acetic acid and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0213] In the developing solvent, the volume ratio of ethyl acetate, formic acid, glacial acetic acid and water is (14-16): 1: 1: 2;

[0214] The color developer is a sulfuric acid ethanol solution; optionally, in the sulfuric acid ethanol solution, the volume percentage of sulfuric acid is 10%.

[0215] In some examples, the method for thin layer chromatography identification of the traditional Chinese medicine preparation Bupleurum for treating leucorrhea comprises the following steps:

[0216] Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution;

[0217] The control medicinal material solution was prepared by mixing water and the control medicinal material Bupleurum respectively, decocting, filtering and taking the filtrate;

[0218] Dissolve saikosaponin a and saikosaponin d reference substances in methanol to prepare reference substance solutions;

[0219] The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, developed with a mixture of ethyl acetate, methanol and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0220] In the developing solvent, the volume ratio of ethyl acetate, methanol and water is (7-9):2:1;

[0221] The color developer is a sulfuric acid solution of p-dimethylaminobenzaldehyde; optionally, in the sulfuric acid solution of p-dimethylaminobenzaldehyde, the volume percentage of p-dimethylaminobenzaldehyde is 2%.

[0222] In some examples, the method for thin layer chromatography identification of dried tangerine peel and schizonepeta spicate in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps:

[0223] Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and ethyl acetate, combining the organic phases, and preparing a test solution;

[0224] The control medicinal materials of dried tangerine peel and schizonepeta spicate were respectively mixed with water, decocted, filtered and the filtrate was collected to prepare a control medicinal material solution;

[0225] The hesperidin reference substance was dissolved in methanol to prepare a reference substance solution;

[0226] The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, first developed with a mixture of ethyl acetate, methanol and water for 4.5 cm, then developed with a mixture of cyclohexane, ethyl acetate, formic acid and water for 8 cm, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm;

[0227] In the developing solvent, the volume ratio of ethyl acetate, methanol and water is 100:(16-18):(12-14); the volume ratio of cyclohexane, ethyl acetate, formic acid and water is (9-11):(9-11):2:5;

[0228] The color developer is an aluminum chloride ethanol solution; optionally, in the aluminum chloride ethanol solution, the volume percentage of aluminum chloride is 5%.

[0229] In some of the examples, the dosage form of the traditional Chinese medicine preparation for treating leucorrhea is selected from one of pills, powders, ointments, tablets, granules, lozenges, capsules, aqueous solutions or tinctures.

[0230] In some examples, the raw materials of the traditional Chinese medicine preparation for treating leukorrhea include the following components in parts by weight:

[0231] 20 parts of stir-fried Atractylodes macrocephala, 20 parts of stir-fried Chinese yam, 10 parts of white peony root with wine, 6 parts of plantain seeds with wine, 6 parts of stir-fried Atractylodes macrocephala with bran, 4 parts of ginseng, 2 parts of liquorice, 1.2 parts of bupleurum, 1 part of dried tangerine peel and 1 part of schizonepeta spicate charcoal;

[0232] In some of the examples, the traditional Chinese medicine preparation for treating leucorrhea is one of the pills, powders, ointments, tablets, granules, lozenges, capsules, aqueous solutions or tinctures of Wandai Decoction.

[0233] Another aspect of the present invention provides the application of the quality detection method of the traditional Chinese medicine preparation for treating leucorrhea as described above in the quality control of the traditional Chinese medicine preparation for treating leucorrhea.

[0234] The following is further described in conjunction with specific embodiments. The raw materials involved in the following specific embodiments, unless otherwise specified, can all be commercially available; the instruments used, unless otherwise specified, can all be commercially available; the processes involved, unless otherwise specified, are all routinely selected by those skilled in the art.

[0235] Example 1

[0236] 1. Study on the fingerprint of Wandai Decoction

[0237] 1 Materials and Instruments

[0238] 1.1 Materials

[0239] Fried Atractylodes macrocephala (Zhejiang, 180401), fried yam (Henan, 180401), white peony root with wine (Anhui, 180501), plantain seed with wine (Jiangxi, 180501), fried Atractylodes macrocephala with bran (Inner Mongolia, 180501), ginseng (Jilin, 170402), licorice (Gansu, 180301), bupleurum (Gansu, 180401), tangerine peel (Hunan, 180201), and charcoal of Schizonepeta tenuifolia (Anhui, 180301).

[0240] 1.2 Instruments

[0241] Heating mantle (Shanghai Huaxing Instrument Factory), electronic balance (Sartorius, BL310), electronic balance (Sartorius, BS224S), electronic balance (METTLER TOLEDO, PL203), electronic balance (METTLER TOLEDO, XP26), rotary evaporator (BUCHI, R-215).

[0242] 2 Experimental methods

[0243] 2.1 Preparation of Wandai Tang Test Solution

[0244] Take 37.50g of stir-fried Atractylodes macrocephala, 37.50g of stir-fried yam, 18.75g of white peony root with wine, 11.25g of Plantain seed with wine, 11.25g of stir-fried Atractylodes lancea with bran, 7.50g of ginseng, 3.75g of licorice, 2.25g of bupleurum, 1.88g of tangerine peel, and 1.88g of Schizonepeta spicate charcoal, add 6 times water, soak for 30min, heat in a round-bottom flask to boil, then decoct for 60min after boiling, filter while hot (300 mesh), and freeze-dry the filtrate.

[0245] Weigh 0.5 g of the freeze-dried powder of the finished soup into a conical flask, add 10 mL of 70% methanol, ultrasonicate for 20 min, shake well, filter through a 0.45 μm filter membrane, and take the filtrate to obtain the final product.

[0246] The preparation methods of control medicinal materials and negative control lyophilized powder are the same as above.

[0247] 2.2 Preparation of reference solution

[0248] Accurately weigh appropriate amounts of gallic acid, paeoniflorin, liquiritin, atractylodes lactone I, atractylodes lactone II, atractylodes lactone III, ammonium glycyrrhizinate, hesperidin, verbascoside, saikosaponin, and liquiritigenin reference substances, and add methanol to prepare a stock solution containing 1 mg of each substance per 1 mL. Take 0.2 mL of the stock solution into a volumetric flask, dilute with methanol to prepare a 200 μg / mL reference substance solution, and obtain the reference substance solution.

[0249] 2.3 Chromatographic conditions

[0250] Chromatographic column: Elite ODS-BP, filler: octadecylsilane bonded silica gel; mobile phase: 0.1% formic acid-water (A)-acetonitrile (B); flow rate: 1.0 mL / min, column temperature: 30°C, detection wavelength: 275 nm, injection volume: 10 μL, elution gradient: as shown in Table 1 below:

[0251] Table 1

[0252]

[0253] 3 Results and discussion

[0254] 3.1 Preparation method investigation

[0255] 3.1.1 Study on decoction time

[0256] The finished decoction medicinal materials under item "2.1" were boiled and then decocted for 30 minutes and 60 minutes respectively, and the extraction yield and HPLC chromatogram were examined. The results showed that the extraction yield of decocting for 30 minutes was 17.6%, and the extraction yield of decocting for 60 minutes was 20.8%.

[0257] Chromatograms of different decoction times are shown in Figure 1 As can be seen from the figure, the chromatograms of decoction for 30 minutes and 60 minutes are basically the same, indicating that its material basis has not changed significantly.

[0258] 3.1.2 Comparison before and after concentration

[0259] After examining the chromatograms of the original solution, concentrated solution, and reconstituted solution after spin drying, the results are as follows: Figure 2 As shown in the figure, there is no significant change in the chromatographic peaks before and after concentration, indicating that the material basis of Wandai Decoction is relatively stable and the material composition will not change significantly before and after concentration.

[0260] 3.1.3 Comparison before and after freeze-drying

[0261] After investigating the changes in the material basis of the water extract before and after freeze-drying, the results are as follows Figure 3 As shown, the results showed that the material basis of the Wandai decoction water extract did not change significantly before and after freeze-drying.

[0262] 3.1.4 Solvent investigation

[0263] (1) Water, different concentrations of ethanol

[0264] Weigh 0.5 g of the lyophilized powder in a conical flask and add H 2 O, 50% ethanol, 70% ethanol, 90% ethanol 10mL, ultrasonic for 20min, shake well, filter through 0.45μm filter membrane, take the filtrate, and analyze by HPLC. Figure 4 As shown, the results showed that the chromatographic peaks of ethanol solutions of different concentrations had significant tailing at 10min-30min. Therefore, 50%-90% ethanol was not suitable as a solvent for Wandai Tang freeze-dried powder.

[0265] (2) Methanol at different concentrations

[0266] Weigh 0.5 g of the lyophilized powder of the finished soup into a conical flask, add 10 mL of 50% methanol, 70% methanol, and 90% methanol, respectively, ultrasonicate for 20 min, shake well, filter through a 0.45 μm filter membrane, take the filtrate, and analyze by HPLC. Figure 5 As shown in the figure, the results show that the gallic acid chromatographic peak of 90% methanol has tailing phenomenon, while the chromatographic peaks of 50% methanol and 70% methanol have no tailing phenomenon. Considering the number of chromatographic peaks and separation degree, 70% methanol was selected as the solvent.

[0267] 3.1.5 Research on the contribution of single medicinal materials

[0268] Referring to the preparation method of the test sample under "2.1", the test solutions of the 10 medicinal materials in the prescription were prepared in turn, and their extract content, extraction yield and contribution rate were investigated. The results are shown in Table 2. The results showed that the top five medicinal materials that contributed the most to the extraction yield in Wandai Decoction were earth-fried Atractylodes macrocephala, ginseng, white peony root with wine, stir-fried Chinese yam, and stir-fried Atractylodes lancea with bran, and the sum of the extraction yield contribution rates reached 95.5%, suggesting that attention should be paid to the quality control of earth-fried Atractylodes macrocephala, ginseng, white peony root with wine, stir-fried Chinese yam, and stir-fried Atractylodes lancea with bran to ensure the stability of product quality.

[0269] Table 2 Extracts and contribution rates of various medicinal materials

[0270]

[0271] 3.2 Optimization of chromatographic conditions

[0272] 3.2.1 Detection wavelength selection

[0273] The chromatograms of Wandai decoction at different detection wavelengths are as follows Figure 6As shown in the figure, the results showed that when 275nm was used as the detection wavelength, the baseline of the chromatogram was stable, more chromatographic peaks were detected, and the peak shape and separation of each chromatographic peak were good, which could more comprehensively reflect the overall appearance of the various components in Wandai Decoction. Therefore, 275nm was selected as the detection wavelength for the HPLC fingerprint study of Wandai Decoction.

[0274] The chromatogram after the chromatographic conditions were optimized is shown in Figure 7 shown.

[0275] 3.3 Main chromatographic peaks and their medicinal properties

[0276] Prepare the test solution with soup, the test solution of ten medicinal materials and the corresponding negative solution according to "2.1" and perform the determination according to the chromatographic conditions under "2.3". The results are as follows Figure 8-Figure 17 By comparing the chromatographic retention behaviors of the main chromatographic peaks of the test solution of the soup and the test solution of the ten medicinal materials, the sources of the 10 main chromatographic peaks were clearly identified.

[0277] 3.3.1 Chromatographic peaks of stir-fried Atractylodes macrocephala

[0278] According to the retention time, the peak of Atractylodes macrocephala mainly appeared in the first 10 min on the HPLC spectrum ( Figure 8 ), peak 1 is partially derived from stir-fried Atractylodes macrocephala.

[0279] 3.3.2 Chromatographic peaks of stir-fried yam

[0280] According to the retention time, peak 6 mainly comes from fried yam ( Fig. 9 ).

[0281] 3.3.3 Chromatographic peaks of Plantago asiatica

[0282] Depend on Fig.10 It can be seen that Peak 10 comes from Plantago asiatica, and Peak 12 partially comes from Plantago asiatica.

[0283] 3.3.4 Chromatographic Peaks of White Peony Root Extract

[0284] Chromatogram of white peony root ( Fig.11 ) It can be seen that Peak 3 and Peak 7 are derived from White Peony Root Wine, and are the exclusive peaks of White Peony Root Wine.

[0285] 3.3.5 Chromatographic peaks of stir-fried Atractylodes lancea

[0286] According to the retention time, peak 6 was partially derived from the fried Atractylodes lancea with bran ( Fig.12 ).

[0287] 3.3.6 Ginseng chromatographic peak attribution

[0288] Depend on Fig.13It can be seen that ginseng basically does not appear in the HPLC chromatogram of Wandai Decoction, which may be due to the small proportion of ginseng in the prescription and the low sensitivity of ginsenosides.

[0289] 3.3.7 Chromatographic peaks of licorice

[0290] Depend on Fig.14 It can be seen that Peak 8, Peak 9 and Peak 14 are derived from licorice and are exclusive peaks of licorice.

[0291] 3.3.8 Chromatographic Peaks of Bupleurum

[0292] From the chromatogram of Bupleurum chinense ( Fig.15 ) It can be seen that the Bupleurum chinense in Wandai Decoction basically does not show any peak, which may be due to the small proportion of its dosage in the whole prescription or the inappropriate sample pretreatment method.

[0293] 3.3.9 Chromatographic Peaks of Tangerine Peel

[0294] According to the retention time, peaks 12 and 13 are derived from tangerine peel and are exclusive peaks of tangerine peel ( Fig.16 ).

[0295] 3.3.10 Chromatographic peaks of Schizonepeta spicata charcoal

[0296] Depend on Fig.17 It can be seen that the charcoal of Schizonepeta spicata in Wandai Decoction basically does not show any peak in the HPLC chromatogram, which may be related to the processing method of Schizonepeta spicata (fried charcoal) and the prescription amount.

[0297] 3.4 Chromatographic peak identification

[0298] Through identification with the retention time and ultraviolet absorption spectrum of the reference substance, a total of 7 chromatographic peaks have been identified, including gallic acid, paeoniflorin, verbascoside, hesperidin, ammonium glycyrrhizinate, atractylodes lactone III, and liquiritin. The specific chromatograms are as follows ( Figure 18-Figure 24 ):

[0299] 4 Summary

[0300] This application investigates the preparation method of Wandai Decoction, chromatographic conditions, attribution of chromatographic peaks of medicinal materials, identification of main chromatographic peaks, and determines the method of fingerprint of Wandai Decoction. There are 14 main chromatographic peaks in total, and 7 chromatographic peaks have been identified so far. The chromatograms of various medicinal materials are summarized in Fig.25 , the summary of the identified chromatographic peaks is shown in Fig.26 .

[0301] 2. Thin layer chromatography study of Wandai decoction

[0302] 1 Materials and Instruments

[0303] 1.1 Materials

[0304] Stir-fried Atractylodes macrocephala (Zhejiang, 180401), stir-fried yam (Henan, 180401), white peony root with wine (Anhui, 180501), plantain seed with wine (Jiangxi, 180501), stir-fried Atractylodes macrocephala with bran (Inner Mongolia, 180501), ginseng (Jilin, 170402), licorice (Gansu, 180301), bupleurum (Gansu, 180401), tangerine peel (Hunan, 180201), schizonepeta spicate charcoal (Anhui, 180301), silica gel G thin layer plate (Merck Chemical Technology Co., Ltd., Qingdao Ocean Chemical Plant), saikosaponin a (China Food and Drug Inspection Institutes, batch number: 200406), saikosaponin d (China Food and Drug Inspection Institutes, batch number: 200505).

[0305] 1.2 Instruments

[0306] Thin layer heater (CAMAG III), UV analyzer (ZF1-I, Jintan Shenglan Instrument Co., Ltd.).

[0307] 2 Experimental methods

[0308] 2.1 Thin layer chromatography identification of Atractylodes macrocephala and Atractylodes lancea

[0309] Take 1.2g of the lyophilized powder with soup, 0.8g of the double negative lyophilized powder of stir-fried Atractylodes macrocephala and stir-fried Atractylodes macrocephala, 0.2g of the lyophilized powder of stir-fried Atractylodes macrocephala, and 0.2g of the lyophilized powder of stir-fried Atractylodes macrocephala, add 10mL of water to dissolve, ultrasonicate for 30 minutes, extract with n-butanol, evaporate the organic layer to dryness, and add 1mL of methanol to dissolve, as the test solution, negative control solution and control medicinal material solution. In addition, take the Atractylodes macrocephala lactone I, II, and III reference substances, add methanol to make a solution containing 1mg per 1ml, as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), take 10μL of the newly prepared solution, spot it on the same silica gel G thin layer plate, use cyclohexane-ethyl acetate-toluene (14:3:3) as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat until the spots are clearly colored, and observe under ultraviolet light (365nm). In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the control medicinal material.

[0310] 2.2 TLC Identification of Yam

[0311] Take 3.6g of lyophilized powder with soup, 1.0g of lyophilized powder of stir-fried yam, and 2.6g of negative lyophilized powder of stir-fried yam, add 20mL of water respectively, dissolve by ultrasonic for 30min, extract with dichloromethane 3 times, 20mL each time, combine the dichloromethane solution, evaporate to dryness, add 1mL of dichloromethane to the residue to dissolve, as the test solution, control medicinal material solution and negative control solution. According to the thin layer chromatography method (General Rule 0502), take 15μL of each of the above 4 solutions, spot them on the same silica gel G thin layer plate, use toluene-acetone (5:1) as the developing solvent, develop, take out, dry, spray with 5% vanillin sulfuric acid solution, and heat at 105℃ until the spots are clearly colored. In the chromatogram of the test sample, the same red spots appear at the corresponding position of the chromatogram of the control medicinal material.

[0312] 2.3 Thin layer chromatography identification of white peony root

[0313] Take 0.71g of lyophilized powder with soup, 0.1g of lyophilized powder with white peony root, and 0.61g of lyophilized powder with white peony root negative, add 20mL of water respectively, ultrasonic for 30 minutes, extract twice with 20mL of ethyl acetate each time, combine ethyl acetate, evaporate to dryness, add 1mL of ethanol to the residue to dissolve, as the test solution, control medicinal material solution and negative control solution. Separately take paeoniflorin and gallic acid reference substances, add ethanol to make a solution containing 1mg per 1mL, as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), take 10μL of each of the above 6 solutions, and spot them on the same silica gel GF254 thin layer plate, use ethyl acetate-methanol-formic acid-water (16:2:1.5:1) as the developing agent, develop, take out, dry, and examine under ultraviolet light (254mn). In the chromatogram of the test substance, spots of the same color appear at the corresponding position of the chromatogram of gallic acid. Spray with 5% vanillin sulfuric acid solution and heat at 105°C until the spots are clearly colored. In the chromatogram of the test sample, the same blue-purple spots appear at the corresponding positions of the chromatograms of paeoniflorin and gallic acid.

[0314] 2.4 Thin layer chromatography identification of Plantago

[0315] Take 1.19g of lyophilized powder with soup, 0.1g of Plantago asiatica lyophilized powder, and 1.09g of Plantago asiatica negative lyophilized powder, add 10mL of water, ultrasonically treat for 30 minutes, cool, extract twice with ether, 20mL each time, discard the ether solution, extract the aqueous solution with water-saturated n-butanol twice, 20mL each time, combine the n-butanol solution, wash the n-butanol solution with n-butanol saturated water twice, 20mL each time, discard the water layer, evaporate to dryness, add 1mL of methanol to the residue to dissolve it, and use it as the test solution, control medicinal material solution and negative control solution. According to the thin layer chromatography method (General Rule 0502), 5 μL of each of the three solutions mentioned above were taken and spotted on the same silica gel G plate, and butyl acetate-methanol-formic acid-water (6:1.5:1.5:1) was used as the developing solvent. The plate was developed, taken out, dried, sprayed with 0.5% vanillin sulfuric acid solution, and heated at 105°C until the spots were clearly colored. The Wandai Decoction showed spots of the same color at the corresponding position of the Plantago seed chromatogram.

[0316] 2.5 Identification of ginseng by thin layer chromatography

[0317] Take 3.56g of lyophilized powder with soup, 0.2g of ginseng lyophilized powder, and 3.36g of ginseng negative lyophilized powder, add 20mL of water respectively, ultrasonic for 30min, extract 3 times with water-saturated n-butanol, 20mL each time, combine the water-saturated n-butanol solution, add 60mL of ammonia test solution, shake well, let it stand for stratification, take the upper layer and wash it 3 times with n-butanol saturated water, 20mL each time, evaporate the upper n-butanol solution to dryness, add 1mL of methanol to dissolve the residue, as the test solution, control medicinal material solution and negative control solution. Take ginsenoside Rb1 reference substance, ginsenoside Re reference substance and ginsenoside Rg1 reference substance, add methanol to make a mixed solution containing 2mg of each in 1mL, as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), 1 to 2 μL of each of the above 7 solutions were taken and spotted on the same silica gel G thin layer plate, and the lower layer solution placed below 10°C in chloroform-ethyl acetate-methanol-water (15:40:22:10) was used as the developing agent. The plate was developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and examined under sunlight and ultraviolet light (365nm). In the chromatogram of the test sample, spots of the same color or fluorescent spots appeared at the corresponding positions of the chromatogram of the reference medicinal material and the chromatogram of the reference sample.

[0318] 2.6 TLC Identification of Licorice

[0319] Take 1.78g of lyophilized powder with soup, 0.05g of lyophilized powder of licorice, and 1.73g of lyophilized powder of licorice negative, add 20mL of water respectively, ultrasonicate for 30min, extract with 40mL of ether, discard the ether solution, extract the water layer with n-butanol 3 times, 20mL each time, combine the n-butanol solution, wash with n-butanol saturated water 3 times, discard the water solution, evaporate the n-butanol solution, add 1mL of methanol to dissolve the residue, and use it as the test solution, control medicinal material solution and negative control solution. Take liquiritigenin, isoliquiritigenin and ammonium glycyrrhizinate reference substances separately, add methanol to make a solution containing 2mg per 1mL, as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), 1 to 2 μL of each of the above 6 solutions were taken and spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water (15:1:1:2) was used as the developing agent. After development, the plate was taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and examined under ultraviolet light (365nm). In the chromatogram of the test sample, a fluorescent spot of the same color appeared at the corresponding position of the chromatogram of the reference medicinal material; and in the chromatogram of the reference sample, the same orange-yellow fluorescent spot appeared at the corresponding position.

[0320] 2.7 TLC Identification of Bupleurum

[0321] Take 1.18g of the lyophilized powder with soup, 0.2g of the lyophilized powder of Bupleurum, and 1.16g of the negative lyophilized powder of Bupleurum, add 20mL of water respectively, ultrasonicate for 30min to dissolve, extract with saturated n-butanol with water for 3 times, 20mL each time, combine the n-butanol solution, wash with ammonia test solution for 2 times, 20mL each time, discard the ammonia test solution, wash with saturated n-butanol water for 2 times, 20mL each time, discard the water solution, evaporate the n-butanol solution, add 1mL of methanol to dissolve the residue, and use it as the test solution, control medicinal material solution and negative control solution. Separately take the reference substances of saikosaponin a and saikosaponin d, add methanol to make a solution containing 0.5mg per 1mL, as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), 5 μL of each of the above 5 solutions was taken and spotted on the same silica gel G thin layer plate, and ethyl acetate-methanol-water (8:2:1) was used as the developing solvent. After development, the plate was taken out, dried, sprayed with 2% p-dimethylaminobenzaldehyde in 40% sulfuric acid solution, heated at 60°C until the spots were clearly colored, and examined under ultraviolet light (365nm). In the chromatogram of the test sample, spots of the same color or fluorescent spots appeared at the corresponding positions of the chromatogram of the reference medicinal material and the chromatogram of the reference sample.

[0322] 2.8 Identification of dried tangerine peel and schizonepeta spicate by charcoal thin

[0323] Take 1.2g of the lyophilized powder of the soup, 0.1g of the lyophilized powder of dried orange peel, 0.5g of the lyophilized powder of Nepeta paniculata, and 1.1g of the double negative lyophilized powder of dried orange peel and Nepeta paniculata, add 20mL of water respectively, dissolve them by ultrasonic for 30min, extract them with ethyl acetate 3 times, 20mL each time, combine the ethyl acetate solution, evaporate to dryness, and dissolve the residue with 1mL of methanol as the test solution, control medicinal material solution and negative control solution. Take another hesperidin reference substance, add methanol to make a 1mL saturated solution as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), 10 μL of the test solution and 5 μL of the reference solution are taken and spotted on the same silica gel G thin layer plate, developed with ethyl acetate-methanol-water (100:17:13) for 4.5 cm, taken out, dried, and then developed with cyclohexane-ethyl acetate-formic acid-water (10:10:2:5) upper layer solution for 8 cm, developed, taken out, dried, sprayed with 5% aluminum chloride ethanol solution, and examined under ultraviolet light (365 nm). In the chromatogram of the test sample, the same fluorescent spot is shown at the corresponding position of the chromatogram of the reference medicinal material.

[0324] 3 Results and discussion

[0325] 3.1 Thin layer chromatography identification of Atractylodes macrocephala and Atractylodes lancea

[0326] like Fig. 27 As shown, in the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatograms of the reference medicinal materials and the Atractylodes lactone I, II, and III reference samples.

[0327] 3.2 TLC Identification of Chinese Yam

[0328] like Fig.28 As shown in the figure, in the chromatogram of the test sample, the same red spot appears at the corresponding position of the chromatogram of the control medicinal material. After being left overnight, the spot becomes clearer.

[0329] 3.3 Thin layer chromatography identification of white peony root

[0330] like Fig.29 As shown, in the chromatogram of the test sample, there are spots of the same color at the corresponding positions in the chromatogram of the reference sample, indicating that Wandai Decoction contains gallic acid and paeoniflorin.

[0331] 3.4 Thin layer chromatography identification of Plantago

[0332] like Fig.30 As shown, Wandai Decoction shows the same purple spots at the corresponding position of the chromatogram of Plantago seed.

[0333] 3.5 Thin layer chromatography identification of ginseng

[0334] In the chromatogram of the test sample, spots of the same color appear at the positions corresponding to ginsenoside Rg1, ginsenoside Rb1, and ginsenoside Re ( Fig.31), indicating that Wandai Decoction contains ginsenoside Rg1, ginsenoside Rb1 and ginsenoside Re.

[0335] 3.6 TLC Identification of Licorice

[0336] like Fig.32 As shown, Wandai Decoction contains liquiritigenin, isoliquiritigenin and ammonium glycyrrhizinate; at the corresponding position of the chromatogram of licorice medicinal materials, Wandai Decoction shows fluorescent spots of the same color.

[0337] 3.7 TLC Identification of Bupleurum

[0338] like Fig.33 As shown, in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of saikosaponin a and Bupleurum medicinal materials, and the sample contains saikosaponin a.

[0339] 3.8 Identification of dried tangerine peel and schizonepeta spicate by charcoal thin layer chromatography

[0340] like Fig.34 As shown, in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions in the chromatograms of the medicinal materials of dried tangerine peel, schizonepeta spicate and the reference sample.

[0341] 4 Summary

[0342] This application investigated the thin layer chromatography identification method of 10 medicinal materials in Wandai Decoction, and the specific situation is summarized in Table 3 below:

[0343] Table 3

[0344] Medicinal Materials Thin layer chromatography identification Atractylodes macrocephala The method is feasible and can detect atractylodes lactones I, II, and III Fried yam The method is feasible White Peony Root The method is feasible and can detect paeoniflorin Plantago The method is feasible Fried Atractylodes macrocephala with bran The method is feasible and can detect atractylodes lactones I, II, and III Ginseng The method is feasible, ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re Licorice The method is feasible and can detect liquiritin, isoliquiritin and ammonium glycyrrhizinate Bupleurum The method is feasible and can detect saikosaponin a Tangerine peel The method is feasible and hesperidin can be detected Nepeta Spike Charcoal The method is feasible and hesperidin can be detected, but pulegone, a pharmacopoeia index, cannot be detected.

[0345] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0346] The above-described embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be construed as limiting the scope of the patent application. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent application shall be subject to the attached claims, and the description may be used to interpret the content of the claims.

Claims

1. A quality detection method for a Chinese medicine preparation for treating leucorrhea, characterized in that: The method comprises the following steps: constructing a fingerprint of the Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography, and identifying the components of the Chinese medicine preparation for treating leukorrhea by thin layer chromatography; the raw materials of the Chinese medicine preparation for treating leukorrhea include the following components: stir-fried Atractylodes macrocephala, stir-fried Chinese yam, white peony root with wine, plantain seed with wine, stir-fried Atractylodes macrocephala with bran, ginseng, liquorice, bupleurum root, dried orange peel and schizonepeta spicate charcoal; The method of constructing the fingerprint of the traditional Chinese medicine preparation for treating leukorrhea by high performance liquid chromatography comprises the following steps: The Chinese herbal medicine for treating leukorrhea is extracted using an extraction solvent to prepare a test solution; The test solution is subjected to high performance liquid chromatography detection to construct a fingerprint of the traditional Chinese medicine preparation for treating leucorrhea; The method of identifying the components of the Chinese medicine preparation for treating leukorrhea by thin layer chromatography comprises the following steps: Take a reference substance, add a dissolving solvent to dissolve it, and prepare a reference substance solution; the reference substance includes at least one of atractylodes lactone I, atractylodes lactone II, atractylodes lactone III, paeoniflorin, gallic acid, ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, liquiritin, isoliquiritigenin, ammonium glycyrrhizinate, saikosaponin a, saikosaponin d, and hesperidin; The traditional Chinese medicine preparation for treating leucorrhea is extracted with water and an organic solvent, and the organic phases are combined to prepare a test solution; Mix water and reference medicinal materials, decoct, filter and collect the filtrate to prepare a reference medicinal material solution; the reference medicinal materials include at least one of stir-fried Atractylodes macrocephala, stir-fried Chinese yam, stir-fried white peony root, stir-fried Plantago asiatica, stir-fried Atractylodes lancea with bran, ginseng, liquorice, bupleurum, dried orange peel and schizonepeta spicate charcoal; The test solution, control medicinal material solution and / or control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a developing agent, dried, developed with a color developing agent, and inspected.

2. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to claim 1, characterized in that, The conditions of the high performance liquid chromatography method include: mobile phase A is formic acid aqueous solution, mobile phase B is acetonitrile, and gradient elution is adopted; The procedure of the gradient elution includes: 0-15min, the volume percentage of the mobile phase B is maintained at 5%; 15min-60min, the volume percentage of the mobile phase B increases from 5% to 30%; 60min-75min, the volume percentage of the mobile phase B increases from 30% to 85%; 75min-85min, the volume percentage of the mobile phase B increases from 85% to 95%; From 85 min to 90 min, the volume percentage of the mobile phase B is maintained at 95%.

3. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to claim 1, characterized in that, When constructing the fingerprint of the traditional Chinese medicine preparation for treating leucorrhea by high performance liquid chromatography, at least one of the following conditions is met: (1) Flow rate: 0.8 mL / min-1.2 mL / min; (2) Column temperature is 25°C-35°C; (3) The detection wavelength is 200nm-400nm; (4) The injection volume is 5 μL-20 μL; (5) In the formic acid aqueous solution, the volume percentage of formic acid is 0.01%-0.15%; (6) The extraction solvent is a methanol-water solution; optionally, the volume percentage of methanol in the methanol-water solution is 50%-90%.

4. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to claim 1, characterized in that, The conditions for identifying the Chinese medicine preparation for treating leucorrhea by thin layer chromatography include: the dissolving agent is at least one of methanol and ethanol; and / or The organic phase is selected from at least one of n-butanol, ethyl acetate, dichloromethane and diethyl ether; and / or The developing agent is at least one selected from the group consisting of cyclohexane, a mixture of ethyl acetate and toluene, a mixture of toluene and acetone, a mixture of ethyl acetate, methanol, formic acid and water, a mixture of chloroform, ethyl acetate, methanol and water, a mixture of ethyl acetate, formic acid, glacial acetic acid and water, a mixture of ethyl acetate, methanol and water, and a mixture of cyclohexane, ethyl acetate, formic acid and water; and / or The color developer is selected from at least one of sulfuric acid ethanol solution, vanillin sulfuric acid solution, p-dimethylaminobenzaldehyde sulfuric acid solution and aluminum chloride ethanol solution.

5. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to claim 4, characterized in that, The method for thin layer chromatography identification of Atractylodes macrocephala and Atractylodes lancea in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps: Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution; The control medicinal material solutions were prepared by mixing water, Atractylodes macrocephala control medicinal material and Atractylodes lancea control medicinal material, decocting, filtering and collecting the filtrate; The reference substances of atractylodes lactone I, atractylodes lactone II and atractylodes lactone III were dissolved in methanol to prepare reference substance solutions; The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, developed with a mixture of cyclohexane, ethyl acetate and toluene as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm; In the developing solvent, the volume ratio of cyclohexane, ethyl acetate and toluene is (14-16):3:3; The color developer is a sulfuric acid ethanol solution; optionally, the volume percentage of sulfuric acid in the sulfuric acid ethanol solution is 10%; The method for thin layer chromatography identification of yam in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps: Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and dichloromethane, combining the organic phases, and preparing a test solution; Mix water and yam control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution; The test solution and the control medicinal material solution are spotted on the same silica gel G thin layer plate respectively, and developed with a mixed solution of toluene and acetone as a developing agent, the silica gel G thin layer plate is dried, and a color developer is used for color development; In the developing agent, the volume ratio of toluene to acetone is (4-6):1; The color developer is a vanillin sulfuric acid solution; optionally, in the vanillin sulfuric acid solution, the volume percentage of vanillin is 5%.

6. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to claim 4, characterized in that, The method for thin layer chromatography identification of white peony root in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps: Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and ethyl acetate, combining the organic phases, and preparing a test solution; Mix water and white peony root control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution; The reference substances of paeoniflorin and gallic acid were dissolved in ethanol to prepare reference substance solutions; The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a mixture of ethyl acetate, methanol, formic acid and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 254 nm; In the developing solvent, the volume ratio of ethyl acetate, methanol, formic acid and water is (15-17):2:1.5:1; The color developer is a vanillin sulfuric acid solution; optionally, the volume percentage of vanillin in the vanillin sulfuric acid solution is 5%; The method for thin layer chromatography identification of Plantago in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps: Using water and ether, and water and n-butanol to extract the traditional Chinese medicine preparation for treating leucorrhea in sequence, combining the organic phases, and preparing a test solution; Mix water and Plantago seed control medicinal material, decoct, filter and take the filtrate to prepare a control medicinal material solution; The test solution and the control medicinal material solution are spotted on the same silica gel G thin layer plate, developed with a mixture of butyl acetate, methanol, formic acid and water as a developing agent, dried, and developed with a color developer; In the developing agent, the volume ratio of butyl acetate, methanol, formic acid and water is (5-7): 1.5: 1.5: 1; The color developer is a vanillin sulfuric acid solution; optionally, in the vanillin sulfuric acid solution, the volume percentage of vanillin is 0.5%.

7. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to claim 4, characterized in that, The method for thin layer chromatography identification of ginseng in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps: Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution; The control medicinal material solution is prepared by mixing water and ginseng control medicinal material respectively, decocting, filtering and taking the filtrate; The reference substances of ginsenoside Rb1, ginsenoside Re and ginsenoside Rg1 were dissolved in methanol to prepare reference substance solutions; The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate, developed with a mixture of chloroform, ethyl acetate, methanol and water as a developing agent, dried, developed with a color developer, and examined under sunlight and ultraviolet light at 365 nm, respectively; In the developing solvent, the volume ratio of chloroform, ethyl acetate, methanol and water is (14-16): (39-41): 22: 10; The color developer is a sulfuric acid ethanol solution; optionally, the volume percentage of sulfuric acid in the sulfuric acid ethanol solution is 10%; The method for thin layer chromatography identification of liquorice in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps: Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution; The control medicinal material solution is prepared by mixing water and ginseng control medicinal material respectively, decocting, filtering and taking the filtrate; The reference substances, liquiritin, isoliquiritin and ammonium glycyrrhizinate, were dissolved in methanol to prepare reference substance solutions; The test solution, the control medicinal material solution and the control substance solution are spotted on the same silica gel G thin layer plate respectively, developed with a mixed solution of ethyl acetate, formic acid, glacial acetic acid and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm; In the developing solvent, the volume ratio of ethyl acetate, formic acid, glacial acetic acid and water is (14-16): 1: 1: 2; The color developer is a sulfuric acid ethanol solution; optionally, in the sulfuric acid ethanol solution, the volume percentage of sulfuric acid is 10%.

8. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to claim 4, characterized in that, The method for thin layer chromatography identification of the Chinese medicinal preparation Bupleurum for treating leucorrhea comprises the following steps: Extracting the traditional Chinese medicine preparation for treating leucorrhea with water and n-butanol, combining the organic phases, and preparing a test solution; The control medicinal material solution was prepared by mixing water and the control medicinal material Bupleurum respectively, decocting, filtering and taking the filtrate; Dissolve saikosaponin a and saikosaponin d reference substances in methanol to prepare reference substance solutions; The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, developed with a mixture of ethyl acetate, methanol and water as a developing agent, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm; In the developing solvent, the volume ratio of ethyl acetate, methanol and water is (7-9):2:1; The color developer is a sulfuric acid solution of p-dimethylaminobenzaldehyde; optionally, the volume percentage of p-dimethylaminobenzaldehyde in the sulfuric acid solution of p-dimethylaminobenzaldehyde is 2%; The method for thin layer chromatography identification of dried orange peel and schizonepeta spicate in the traditional Chinese medicine preparation for treating leukorrhea comprises the following steps: Extracting the traditional Chinese medicine preparation for treating leukorrhea with water and ethyl acetate, combining the organic phases, and preparing a test solution; The control medicinal material solutions were prepared by mixing water with dried orange peel and schizonepeta spicata control medicinal materials respectively, boiling them, filtering and collecting the filtrate; The hesperidin reference substance was dissolved in methanol to prepare a reference substance solution; The test solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate, first developed with a mixture of ethyl acetate, methanol and water for 4.5 cm, then developed with a mixture of cyclohexane, ethyl acetate, formic acid and water for 8 cm, dried, developed with a color developer, and examined under an ultraviolet lamp at 365 nm; In the developing solvent, the volume ratio of ethyl acetate, methanol and water is 100:(16-18):(12-14); the volume ratio of cyclohexane, ethyl acetate, formic acid and water is (9-11):(9-11):2:5; The color developer is an aluminum chloride ethanol solution; optionally, in the aluminum chloride ethanol solution, the volume percentage of aluminum chloride is 5%.

9. The quality detection method of the Chinese medicine preparation for treating leucorrhea according to any one of claims 1 to 8, characterized in that: The raw materials of the Chinese medicine preparation for treating leukorrhea include the following components in parts by weight: 20 parts of stir-fried Atractylodes macrocephala, 20 parts of stir-fried Chinese yam, 10 parts of white peony root with wine, 6 parts of plantain seeds with wine, 6 parts of stir-fried Atractylodes macrocephala with bran, 4 parts of ginseng, 2 parts of liquorice, 1.2 parts of bupleurum, 1 part of dried tangerine peel and 1 part of schizonepeta spicate charcoal; Optionally, the Chinese medicine preparation for treating leucorrhea is one of the pills, powders, pastes, tablets, granules, lozenges, capsules, aqueous solutions or tinctures of Wandai Decoction.

10. Application of the quality detection method of the traditional Chinese medicine preparation for treating leucorrhea according to any one of claims 1 to 9 in the quality control of the traditional Chinese medicine preparation for treating leucorrhea.

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