Quality detection method of capsules for purging liver fire and soothing nerves

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a quality detection method of capsules for purging liver-fire and soothing nerves, which comprises the following steps: identifying gentiopicroside, geniposide, scutellaria baicalensis, polygala tenuifolia, angelica sinensis and rhizoma alismatis in the capsules for purging liver-fire and soothing nerves by adopting a thin layer chromatography; and simultaneously detecting the contents of gentiopicroside, geniposide and baicalin by adopting a high performance liquid chromatography. The detection method of the liver-fire-purging and nerve-calming capsule has the advantages of being rapid in analysis, simple to operate, high in accuracy, wide in adaptability and the like, and is beneficial to establishment and overall evaluation of quality standards of the liver-fire-purging and nerve-calming capsule.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a quality testing method for liver-clearing and calming capsules. Background Technology

[0002] Xiegan Anshen Capsules are a modified version of Xiegan Anshen Pills. The current quality standard is listed in Volume 64 of the National New Drug Approval Standards, standard number WS3-620(Z-103)-2004(Z). The prescription consists of Gentiana scabra, Scutellaria baicalensis, Gardenia jasminoides (ginger-processed), Mother-of-Pearl, Oyster shell, Dragon bone, Arborvitae seed, Ziziphus jujuba seed (fried), Polygala tenuifolia (heart removed, licorice-processed), Angelica sinensis, Rehmannia glutinosa, Ophiopogon japonicus, Tribulus terrestris (thorns removed, salt-processed), Poria cocos, Plantago asiatica (salt-processed), Alisma plantago-aquatica (salt-processed), and Glycyrrhiza uralensis. Poria cocos and Alisma plantago-aquatica are powdered and prepared for later use. Mother-of-Pearl, Dragon bone, and Oyster shell are powdered and decocted for a long time to make an extract. The remaining 12 herbs, including Gentiana scabra, are decocted together to make an extract. The two extracts are mixed with the powdered Poria cocos and Alisma plantago-aquatica, dried, pulverized, sieved, and then filled into capsules. It is mainly used for insomnia, restlessness, and neurasthenia caused by excessive liver fire and mental agitation.

[0003] The existing quality standard for Xiegan Anshen Capsules (WS3-620(Z-103)-2004(Z)) includes microscopic identification of Poria cocos and Alisma plantago-aquatica, thin-layer chromatography identification of gentiopicrin, geniposide, baicalin, and Angelica sinensis, and content determination of gentiopicrin. Compared with the quality standard for Xiegan Anshen Pills (Chinese Pharmacopoeia 2020 Edition, Part I), the existing quality standard for Xiegan Anshen Capsules lacks thin-layer chromatography identification of Polygala tenuifolia, and uses a single reference standard as a reference for thin-layer chromatography of Gardenia jasminoides and Scutellaria baicalensis, which cannot comprehensively control the quality of the drug; the content limit is much higher than that of Xiegan Anshen Pills.

[0004] Therefore, the existing quality standards and testing methods have certain limitations in the quality control of "multi-component liver-clearing and calming capsules". New testing methods need to be developed to make the quality control of the drug safer, more uniform and effective. Summary of the Invention

[0005] The purpose of this invention is to provide a new quality testing method for Xiegan Anshen Capsules. By analyzing the components in the formulation, a new thin-layer chromatography identification method is added for multiple medicinal materials such as gentiopicrin, geniposide, scutellaria baicalensis, polygala tenuifolia, and alisma plantago-aquatica, thereby improving the accuracy and comprehensiveness of the quality control of Xiegan Anshen Capsules.

[0006] This invention provides a quality testing method for Xiegan Anshen Capsules, wherein the raw materials of Xiegan Anshen Capsules include: gentian, mother-of-pearl, scutellaria, gardenia, tribulus terrestris, oyster shell, dragon bone, poria cocos, plantain seed, alisma plantago-aquatica, jujube seed, arborvitae seed, polygala tenuifolia, rehmannia glutinosa, ophiopogon japonicus, angelica sinensis, and licorice.

[0007] The quality testing method for the liver-clearing and calming capsules includes at least one of the following methods:

[0008] (1) Thin-layer chromatography detection method for gentiopicrin and geniposide;

[0009] (2) Thin-layer chromatography detection method for Scutellaria baicalensis;

[0010] (3) Thin-layer chromatography detection method for Polygala tenuifolia;

[0011] The thin-layer chromatography detection method for gentiopicrin and geniposide includes the following steps:

[0012] Prepare test solution A of liver-clearing and calming capsules, gentiopicroside reference solution and geniposide reference solution respectively. Spot the test solution A, gentiopicroside reference solution and geniposide reference solution on a glass pre-made silica gel G thin layer plate. Develop with a mixed solvent of dichloromethane, methanol and water in a volume ratio of (25-35):(8-10):(2-4) as the developing solvent. After color development with sulfuric acid ethanol solution, perform the first inspection.

[0013] The preparation of the test solution A includes the following steps: taking the contents of the liver-clearing and calming capsules and mixing them with a first organic solvent for extraction, and using the resulting extract directly as the test solution A; further, the first organic solvent is one or more of methanol, ethanol and ethyl acetate;

[0014] The thin-layer chromatography detection method for Scutellaria baicalensis includes the following steps:

[0015] Prepare the test solution B of Xiegan Anshen Capsules and the reference medicinal material solution of Scutellaria baicalensis. Spot the test solution B and the reference medicinal material solution of Scutellaria baicalensis on silica gel G pre-prepared thin layer plates containing 0.1%-10% sodium hydroxide. Develop the plates with a mixed solvent of ethyl acetate, butanone, formic acid and water in a volume ratio of (4-6):(2-4):1:1 as the developing solvent. After color development, perform a second inspection.

[0016] The preparation of the test solution B includes the following steps: extracting the contents of the liver-clearing and calming capsules with a second organic solvent to obtain filtrate 1 and residue 1; extracting the residue 1 with a third organic solvent to obtain filtrate 2; concentrating and diluting the filtrate 2; and extracting it with ethyl acetate to obtain aqueous solution 1 and organic phase 1; concentrating and diluting the organic phase 1 to prepare the test solution B; further, the second or third organic solvent is one or more of diethyl ether, methanol, ethanol, and ethyl acetate;

[0017] The thin-layer chromatography detection method for Polygala tenuifolia includes the following steps:

[0018] The test solution C of Xiegan Anshen Capsules and the reference solution of Polygala tenuifolia were prepared separately. The mixture of chloroform, acetone and glacial acetic acid in a volume ratio of (8-10):(4-6):(0.1-0.3) was used as the developing solvent. After color development, the third inspection was performed.

[0019] The preparation method of the test solution C includes the following steps: acidifying and hydrolyzing the aqueous solution 1, extracting it with a fourth organic solvent to obtain an organic phase 2, concentrating and diluting the organic phase 2, and preparing the test solution C; further, the fourth organic solvent is one or more of chloroform, dichloromethane, carbon tetrachloride, and ethyl acetate.

[0020] Furthermore, in the thin-layer chromatography detection method for gentiopicrin and geniposide, the concentration of the sulfuric acid ethanol solution is 5%-15%.

[0021] Furthermore, the concentration of the sulfuric acid ethanol solution is 10%.

[0022] Furthermore, in the thin-layer chromatography detection method for Scutellaria baicalensis, the thin-layer plate is a silica gel G pre-prepared thin-layer plate containing 0.5%-5% sodium hydroxide.

[0023] Furthermore, the thin-layer plate is a pre-fabricated thin-layer plate of silica gel G containing 1% sodium hydroxide.

[0024] Furthermore, in the thin-layer chromatography detection method for Polygala tenuifolia, the volume ratio of chloroform, acetone, and glacial acetic acid is 9:5:0.2.

[0025] In existing technologies, the pretreatment methods for thin-layer chromatography detection of gentiopicrin and geniposide are relatively cumbersome, especially requiring purification treatment, such as purification through a neutral alumina column. However, this invention simplifies the pretreatment process while still meeting the detection requirements by improving the thin-layer plate and the colorimetric reagent.

[0026] In addition, in the thin-layer chromatography detection of Scutellaria baicalensis, when using the standard WS3-620(Z-103)-2004(Z) to determine Scutellaria baicalensis (the developing solvent is ethyl acetate-butanone-formic acid-water (5:3:1:1)), the thin-layer chromatogram has a severe tailing phenomenon and is not obvious, which may lead to misjudgment when used for identification. However, the inventor accidentally discovered that after changing the thin-layer plate from the silica gel G pre-prepared thin-layer plate with 4% sodium acetate to the silica gel G pre-prepared thin-layer plate containing 1% sodium hydroxide, the spots in the thin-layer chromatogram are clear and there is no tailing phenomenon.

[0027] In the thin-layer chromatography detection of Polygala tenuifolia, the inventor found through experiments that when using the thin-layer chromatography developing system for the identification of Xiegan Anshen Pills in the 2020 Edition of Chinese Pharmacopoeia (chloroform-acetone-glacial acetic acid (9:1:0.2)), the Rf value of polygalaxanthone III is relatively low (<0.2), which does not meet the requirements for Rf in general thin-layer identification. If there are other interferences, it is easy to misjudge. Therefore, the inventor improved it. However, through a large number of experiments, it was found that whether the developing agent with greater polarity was replaced or the content of glacial acetic acid was increased, the requirements could not be met. Only when using chloroform-acetone-glacial acetic acid (9:5:0.2) as the developing agent could the detection requirements be met, and the Rf was 0.5.

[0028] In the present invention, the preparation methods of the gentiopicroside reference substance solution and the gardenoside reference substance solution include the following steps: respectively take the gentiopicroside reference substance and the gardenoside reference substance, and dissolve them in a polar organic solvent to obtain the gentiopicroside reference substance solution and the gardenoside reference substance solution.

[0029] Further, the polar organic solvent is one or more of methanol, ethanol, and ethyl acetate.

[0030] In the present invention, the preparation method of the Scutellaria baicalensis Georgi reference medicinal material solution includes the following steps: mix the Scutellaria baicalensis Georgi reference medicinal material with ether for extraction to obtain residue 2, mix the residue 2 with methanol for extraction to obtain filtrate 3, concentrate and dilute the filtrate 3, then mix it with ethyl acetate for extraction to obtain organic phase 3, and then concentrate, dilute, and prepare the Scutellaria baicalensis Georgi reference medicinal material solution.

[0031] In the present invention, the preparation method of the Polygala tenuifolia reference substance solution includes the following steps: take the polygalaxanthone III reference substance and dissolve it in a polar organic solvent to obtain the Polygala tenuifolia reference substance solution. 3]

[0032] Further, the polar organic solvent is one or more of methanol, ethanol, and ethyl acetate.

[0033] In the present invention, the detection method further includes the thin-layer chromatography detection method of Angelica sinensis and the thin-layer chromatography detection method of Alisma orientale.

[0034] The thin-layer chromatography detection method of Angelica sinensis includes the following steps: respectively prepare the test solution D of Xiegan Anshen Capsule and the Angelica sinensis reference medicinal material solution, spot the test solution D and the Angelica sinensis reference medicinal material solution on a prefabricated silica gel G thin-layer plate, develop with a mixed solvent of n-hexane and ethyl acetate with a volume ratio of (8-10):1 as the developing agent, and conduct the fourth inspection.

[0035] The thin-layer chromatography detection method for Alisma plantago-aquatica includes the following steps: developing the test solution E of Xiegan Anshen Capsules and the reference herb solution of Alisma plantago-aquatica with a volume ratio of 1:1 using a mixed solvent of n-hexane and ethyl acetate as the developing solvent, and performing a fifth inspection after color development with a colorimetric reagent.

[0036] In this invention, the preparation of the test solution D includes the following steps: concentrating and diluting the filtrate 1 to prepare the test solution D;

[0037] The preparation of the Angelica sinensis reference herb solution includes the following steps: mixing the Angelica sinensis reference herb with a second organic solvent for extraction to obtain filtrate 4, concentrating and diluting the filtrate 4, and preparing the Angelica sinensis reference herb solution;

[0038] The preparation of the test solution E includes the following steps: taking the test solution A as the test solution E;

[0039] The preparation of the Alisma plantago-aquatica reference herb solution includes the following steps: Alisma plantago-aquatica reference herb is mixed with ether for extraction to obtain filtrate 5, and the filtrate 5 is concentrated, diluted, and prepared into the Alisma plantago-aquatica reference herb solution.

[0040] In this invention, the detection method further includes a high-performance liquid chromatography method for simultaneously determining the contents of gentiopicrin, geniposide, and baicalin, comprising the following steps:

[0041] (1) Sample pretreatment: Mix the contents of Xiegan Anshen capsules with 50-100% methanol solution to extract the sample solution to be tested; preferably 70% methanol solution;

[0042] (2) Chromatographic analysis: The sample solution to be tested was analyzed and determined by high performance liquid chromatography;

[0043] The elution process used octadecylsilane-bonded silica gel as the packing material; methanol as mobile phase A and 0.2% phosphoric acid solution as mobile phase B; the gradient elution program was as follows:

[0044]

[0046] The beneficial effects of the technical solution of this invention:

[0047] 1) Identification of multiple components enables comprehensive control of drug quality, resulting in safer, more uniform, and more effective quality control;

[0048] 2) Optimize the thin-layer chromatography identification of gentiopicrin and geniposide, simplify the pretreatment, and improve the operability and accuracy of thin-layer chromatography identification;

[0049] 3) The pretreatment process for Angelica sinensis, Scutellaria baicalensis, and Polygala tenuifolia samples was standardized, simplifying the operation;

[0050] 4) Add Scutellaria baicalensis as a reference material to improve the quality control capability of thin-layer chromatography identification;

[0051] 5) The HPLC method can simultaneously determine the contents of gentiopicrin, geniposide, and baicalin, simplifying the operation and reducing costs. Attached Figure Description

[0052] Figure 1 Thin-layer chromatograms of gentiopicrin and geniposide (Standard WS3-620(Z-103)-2004(Z));

[0053] Figure 2 : Thin-layer chromatograms of gentiopicrin and geniposide (condition A);

[0054] Figure 3 : Thin-layer chromatograms of gentiopicrin and geniposide (condition B);

[0055] Figure 4 : Thin-layer chromatograms of gentiopicrin and geniposide (condition C);

[0056] Figure 5 Thin-layer chromatogram of Angelica sinensis;

[0057] Figure 6 : Thin-layer chromatogram of Alisma plantago-aquatica (developing solvent system I);

[0058] Figure 7 : Thin-layer chromatogram of Alisma plantago-aquatica (developing solvent system II);

[0059] Figure 8 Thin-layer chromatograms of Scutellaria baicalensis (developing solvent system I, A is a thin-layer chromatogram under a silica gel G pre-prepared thin-layer plate containing 1% sodium hydroxide; B is a thin-layer chromatogram under a silica gel G pre-prepared thin-layer plate containing 4% sodium acetate).

[0060] Figure 9 Thin-layer chromatogram of Scutellaria baicalensis (developing solvent system II);

[0061] Figure 10 : Thin-layer chromatogram of Polygala tenuifolia (developing solvent system I);

[0062] Figure 11 : Thin-layer chromatogram of Polygala tenuifolia (developing solvent system II);

[0063] Figure 12 Thin-layer chromatogram of Polygala tenuifolia (developing solvent system) );

[0064] Figure 13 Thin-layer chromatogram of Polygala tenuifolia (developing solvent system) );

[0065] Figure 14 : Liquid chromatograms of gentiopicrin, geniposide, and baicalin;

[0066] Figure 15 Content of gentiopicrin, geniposide, and baicalin (determination method of this invention);

[0067] Figure 16 Content of gentiopicrin, geniposide, and baicalin (Standard (WS3-620-(Z-103)-2004Z)). Detailed Implementation

[0068] The present invention will be further described in detail below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of the disclosure of the present invention.

[0069] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0070] The term "preferred" in this invention refers to an embodiment of the invention that provides certain beneficial effects under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the description of one or more preferred embodiments does not imply that other embodiments are unavailable, nor is it intended to exclude other embodiments from the scope of the invention. That is, in this invention, "preferred" merely describes a more effective implementation or embodiment, but does not constitute a limitation on the scope of protection of the invention.

[0071] In this invention, terms such as "further" and "even further" are used for descriptive purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this invention.

[0072] In this invention, "at least one" means one or more, such as one, two or more. "Multiple" or "several" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0073] When a numerical range is disclosed in this invention, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Further, when the range refers to an integer, it includes every integer between the minimum and maximum values ​​of the range. Moreover, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0074] Unless otherwise specified, all steps of this invention may be performed sequentially or randomly. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For example, the method may also include step (c), indicating that step (c) may be added to the method in any order. For example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0075] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature.

[0076] In this invention, the terms "comprising," "including," "containing," "having," or other variations are intended to cover non-closed inclusion, and no distinction is made between these terms. The term "comprising" means that other steps and ingredients may be added without affecting the final result.

[0077] The Liver-Clearing and Tranquilizing Capsules are made from 17 medicinal materials, including gentian, mother-of-pearl, scutellaria, gardenia (ginger-processed), tribulus terrestris (thorn removed and salt-processed), oyster shell, dragon bone, poria cocos, plantain seed (salt-processed), alisma plantago-aquatica (salt-processed), jujube seed (fried), arborvitae seed, polygala tenuifolia (heart removed and licorice-processed), rehmannia root, ophiopogon japonicus, angelica sinensis, and licorice.

[0078] Thin-layer chromatography (TLC) involves coating a suitable stationary phase onto a glass plate, plastic, or aluminum substrate to form a uniform thin layer. After spotting and development, the ratio shift (Rf) is compared with the ratio shift (Rf) of a suitable reference chromatogram obtained using the same method. This method is used for drug identification, impurity detection, or content determination. As a crucial experimental technique for the rapid separation and qualitative analysis of small amounts of substances, TLC features short development time, strong separation capability, and high sensitivity, making it a vital analytical tool for the quality control of traditional Chinese medicine.

[0079] To improve the accuracy and comprehensiveness of quality control for Xiegan Anshen Capsules, this invention provides a thin-layer chromatography method for detecting multiple medicinal materials such as gentian, gardenia, scutellaria, and polygala in Xiegan Anshen Capsules.

[0080] The thin-layer chromatography detection method for gentiopicrin and geniposide includes the following steps:

[0081] Prepare test solution A of liver-clearing and calming capsules, gentiopicroside reference solution and geniposide reference solution respectively. Spot the test solution A, gentiopicroside reference solution and geniposide reference solution on a glass pre-made silica gel G thin layer plate. Develop with a mixed solvent of dichloromethane, methanol and water in a volume ratio of (25-35):(8-10):(2-4) as the developing solvent. After color development with sulfuric acid ethanol solution, perform the first inspection.

[0082] The preparation of the test solution A includes the following steps: taking the contents of Xiegan Anshen capsules and mixing them with a first organic solvent for extraction, and using the resulting extract directly as test solution A;

[0083] The thin-layer chromatography detection method for Scutellaria baicalensis includes the following steps:

[0084] Prepare the test solution B of Xiegan Anshen Capsules and the reference medicinal material solution of Scutellaria baicalensis. Spot the test solution B and the reference medicinal material solution of Scutellaria baicalensis on silica gel G pre-prepared thin layer plates containing 0.1%-10% sodium hydroxide. Develop the plates with a mixed solvent of ethyl acetate, butanone, formic acid and water in a volume ratio of (4-6):(2-4):1:1 as the developing solvent. After color development, perform a second inspection.

[0085] The preparation of the test solution B includes the following steps: the contents of the liver-clearing and calming capsules are mixed with a second organic solvent for extraction to obtain filtrate 1 and residue 1; the residue 1 is mixed with a third organic solvent for extraction to obtain filtrate 2; the filtrate 2 is concentrated and diluted, and then extracted with ethyl acetate to obtain aqueous solution 1 and organic phase 1; the organic phase 1 is concentrated and diluted to prepare the test solution B.

[0086] The thin-layer chromatography detection method for Polygala tenuifolia includes the following steps:

[0087] The test solution C of Xiegan Anshen Capsules and the reference solution of Polygala tenuifolia were prepared separately. The mixture of chloroform, acetone and glacial acetic acid in a volume ratio of (8-10):(4-6):(0.1-0.3) was used as the developing solvent. After color development, the third inspection was performed.

[0088] The preparation method of the test solution C includes the following steps: acidifying and hydrolyzing the aqueous solution 1, extracting it with a fourth organic solvent to obtain an organic phase 2, concentrating and diluting the organic phase 2, and preparing the test solution C.

[0089] This can be understood as meaning that after the contents of the liver-clearing and calming capsules are mixed with the first organic solvent for extraction, no purification is required to meet the detection requirements.

[0090] It can be understood that the volume ratio of the mixed solvent of dichloromethane, methanol and water can be, but is not limited to, 30:8:3, 30:9:3, 30:10:3, 25:9:3, 26:9:3, 27:9:3, 28:9:3, 29:9:3, 30:9:3, 31:9:3, 32:9:3, 33:9:3, 34:9:3, 35:9:3, 30:9:2, 30:9:4, etc.; preferably, the volume ratio of the mixed solvent of dichloromethane, methanol and water is 30:9:3.

[0091] It can be understood that the volume ratio of the mixed solvent of ethyl acetate, butanone, formic acid and water can be, but is not limited to, 4:3:1:1, 5:3:1:1, 6:3:1:1, 5:2:1:1, 5:4:1:1, etc.; preferably, the volume ratio of the mixed solvent of ethyl acetate, butanone, formic acid and water is 5:3:1:1.

[0092] It can be understood that the volume ratio of the mixed solvent of chloroform, acetone and glacial acetic acid can be, but is not limited to, 8:5:0.2, 9:5:0.2, 10:5:0.2, 9:4:0.2, 9:6:0.2, 9:5:0.1, 9:5:0.3, etc.; preferably, the volume ratio of the mixed solvent of chloroform, acetone and glacial acetic acid is 9:5:0.2.

[0093] It can be understood that the concentration includes, but is not limited to: atmospheric pressure evaporation, vacuum concentration, thin film evaporation, freeze drying, membrane separation, etc.

[0094] This can be understood as the dilution specifically referring to the process of dissolving the concentrated substance in an organic solvent.

[0095] In some embodiments, in the thin-layer chromatography detection method for gentiopicrin and geniposide, the colorimetric reagent is a 5%-15% sulfuric acid ethanol solution.

[0096] It can be understood that the concentration of the sulfuric acid ethanol solution can be 5%, 6%, 7%, 8%, 9%, or 10%; preferably, the concentration of the sulfuric acid ethanol solution is 10%.

[0097] It can be understood that the concentration of sodium hydroxide can be 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, etc.; preferably, the concentration of sodium hydroxide is 1%.

[0098] It can be understood that the solvent extraction involved in this invention can be performed 1, 2, 3, 4, or 5 times.

[0099] The detection methods also include thin-layer chromatography detection methods for Angelica sinensis and Alisma plantago-aquatica.

[0100] The thin-layer chromatography detection method for Angelica sinensis includes the following steps: preparing the test solution D of Xiegan Anshen Capsules and the reference medicinal material solution of Angelica sinensis respectively; spotting the test solution D and the reference medicinal material solution of Angelica sinensis on a glass prefabricated silica gel G thin-layer plate; developing the plate with a mixed solvent of n-hexane and ethyl acetate in a volume ratio of (8-10):1 as the developing solvent; and performing a fourth inspection.

[0101] The thin-layer chromatography detection method for Alisma plantago-aquatica includes the following steps:

[0102] The test solution E of Xiegan Anshen Capsules and the reference herb solution of Zexie were developed and colored using a mixed solvent of n-hexane and ethyl acetate at a volume ratio of 1:1 as the developing solvent and then subjected to the fifth inspection.

[0103] It can be understood that the volume ratio of the mixed solvent of n-hexane and ethyl acetate includes, but is not limited to, 8:1, 9:1, 10:1, etc.; preferably, the volume ratio of the mixed solvent of n-hexane and ethyl acetate is 9:1.

[0104] In this invention, the color development step of the color developer includes: adding the color developer to a thin-layer plate and heating it at a certain temperature until the spots are clear.

[0105] In this invention, the colorimetric agents include, but are not limited to, sulfuric acid ethanol solution, vanillin sulfuric acid solution, vanillic acid sulfuric acid solution, aluminum chloride ethanol solution, and ferric chloride ethanol solution.

[0106] It can be understood that the colorimetric agent can be one or more colorimetric agents; when multiple colorimetric agents are used, they can be mixed or used sequentially, and the order is not limited.

[0107] In some embodiments, the color developer treatment temperature is 100°C-110°C.

[0108] It can be understood that the temperature of the color development treatment includes, but is not limited to, 100℃, 102℃, 104℃, 105℃, 108℃, 110℃, etc.; preferably, the temperature of the color development treatment is 105℃.

[0109] In this invention, the inspection light source is an ultraviolet lamp and / or a fluorescent lamp.

[0110] This can be understood as including, but not limited to, conventional incandescent lamps.

[0111] In some implementation methods, for ease of operation and cost savings, the preparation of the test solution in the thin-layer chromatography detection method for Angelica sinensis, Scutellaria baicalensis, and Polygala tenuifolia can be carried out in the following order:

[0112] (1) The contents of Xiegan Anshen capsules are mixed with a second organic solvent for extraction to obtain filtrate 1 and residue 1. The residue 1 is mixed with a third organic solvent for extraction to obtain filtrate 2. The filtrate 2 is concentrated and diluted, and then extracted with ethyl acetate to obtain aqueous solution 1 and organic phase 1. The organic phase 1 is concentrated and diluted to prepare the test solution B; used for thin-layer chromatography detection of Scutellaria baicalensis.

[0113] (2) After acidifying and hydrolyzing the aqueous solution 1, extract it with a fourth organic solvent to obtain organic phase 2. Concentrate and dilute the organic phase 2 to prepare the test solution C; for thin-layer chromatography detection of Polygala tenuifolia.

[0114] (3) Concentrate and dilute the filtrate 1 to prepare the test solution D; for the chromatographic detection of Angelica sinensis.

[0115] The above-mentioned thin-layer chromatography method can identify the five medicinal materials in the Xiegan Anshen Capsules: gentian, gardenia, scutellaria, alisma, polygala, and angelica. The corresponding thin-layer chromatographic spots are clear, which improves the accuracy and comprehensiveness of the quality control of the Xiegan Anshen Capsules and is more conducive to the establishment and overall evaluation of the quality standards of the Xiegan Anshen Capsules.

[0116] The detection method further includes a high-performance liquid chromatography method for simultaneously determining the contents of gentiopicrin, geniposide, and baicalin, comprising the following steps:

[0117] (1) Sample pretreatment: The contents of Xiegan Anshen capsules were mixed with 50-100% methanol solution for extraction to obtain the sample solution to be tested;

[0118] (2) Chromatographic analysis: The sample solution to be tested was analyzed and determined by high performance liquid chromatography;

[0119] The elution process used octadecylsilane-bonded silica gel as the packing material; methanol as mobile phase A and 0.2% phosphoric acid solution as mobile phase B; the gradient elution program was as follows:

[0120]

[0122] It can be understood that the concentration of the methanol solution can be 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, and 100%, etc., and preferably, the concentration of the methanol solution is 70%.

[0123] It can be understood that the extraction methods include, but are not limited to, immersion, ultrasonic extraction, and hot reflux, and one or more extraction methods can be used in combination; preferably, the present invention uses a combination of ultrasonic extraction and hot reflux for extraction.

[0124] Understandably, octadecyl bonded silica columns are generally chosen, such as: Agilent ZORBAX StableBond C18 4.6×250mm 5μm column, Waters X-Bridge Cl8 4.6x250 mm 5μm column, and SHIMADZU Shim-pack GIST 4.6x250mm 5μm column.

[0125] The high-performance liquid chromatography method described above allows for the simultaneous determination of the contents of gentiopicrin, geniposide, and baicalin, avoiding the introduction of other medicinal materials that may affect the identification, simplifying the operation and reducing costs.

[0126] The following description, in conjunction with specific embodiments and comparative examples, further illustrates the point:

[0127] Example 1: Preparation of Liver-Clearing and Tranquilizing Capsules

[0128] Prescription: Gentian root 83.33g, Mother-of-pearl 555.55g, Scutellaria baicalensis 83.33g, Gardenia jasminoides (ginger-processed) 83.33g, Tribulus terrestris (thorns removed, salt-processed) 83.33g, Oyster shell 138.89g, Dragon bone 138.89g, Poria cocos 83.33g, Plantago asiatica (salt-processed) 83.33g, Alisma plantago-aquatica (salt-processed) 83.33g, Ziziphus jujuba seed (fried) 138.89g, Platycladus orientalis seed 83.33g, Polygala tenuifolia (heart removed, licorice-processed) 83.33g, Rehmannia glutinosa 83.33g, Ophiopogon japonicus 83.33g, Angelica sinensis 83.33g, Licorice 27.78g

[0129] The above seventeen ingredients are prepared as follows: Poria cocos and Alisma plantago-aquatica are pulverized into a fine powder and sieved; Pearl powder, oyster shell, and dragon bone are decocted four times with water, each time for 2 hours, and the decoctions are combined and concentrated into a thick paste; the remaining twelve ingredients, including Gentiana scabra, are decocted three times with water, each time for 1 hour, and the decoctions are combined and concentrated under reduced pressure to a clear paste with a relative density of 1.10 (60℃). Ethanol is added to make the alcohol content 70%, stirred well, allowed to stand, filtered, and the ethanol is recovered from the filtrate and concentrated into a thick paste. This paste is combined with the thick paste of the three ingredients, including pearl powder, and the above-mentioned medicinal powder is added, mixed well, dried, pulverized, sieved, and filled into capsules to make 1000 capsules.

[0130] The Liver-Clearing and Tranquilizing Capsules are an exclusive product of Chengdu Betterdenuo Pharmaceutical Co., Ltd., and a total of 15 batches were collected, as detailed in Table 1.

[0131] Table 1 Sample Information Table for Xiegan Anshen Capsules

[0132]

[0133] Example 2: Thin-layer chromatographic identification of gentiopicrin and geniposide

[0134] 1 Experimental Methods

[0135] 1.1 Preparation of test solution

[0136] Preparation of test solution: Take 2g of the contents of Xiegan Anshen capsules, add 10mL of methanol, sonicate for 30 minutes, let stand, and use the supernatant as the test solution.

[0137] 1.2 Preparation of reference solution

[0138] Take gentiopicrin and geniposide reference standards, and prepare solutions containing 1 mg per 1 mL with methanol, as reference solutions.

[0139] 1.3 Screening of Thin-Layer Chromatography Conditions

[0140] 1.3.1 Condition A

[0141] Take 10 μL of each of the three solutions 1.1 and 1.2 and spot them separately on a glass pre-prepared silica gel G thin-layer plate. Use the lower layer solution of dichloromethane-methanol-water (30:9:3) placed below 10°C as the developing solvent. After development, remove the plate, air dry it, spray it with 10% sulfuric acid ethanol solution, and heat it at 105°C until the spots are clearly visible. Examine it under ultraviolet light (365 nm).

[0142] 1.3.2 Condition B

[0143] Take 10 μL of each of the three solutions 1.1 and 1.2 and spot them separately on a glass pre-prepared silica gel G thin-layer plate. Use ethyl acetate-methanol-water (22:2:1) as the developing solvent. After development, remove the plate, air dry it, spray it with 10% sulfuric acid ethanol solution, and heat it at 105℃ until the spots are clearly visible. Examine it under ultraviolet light (365 nm).

[0144] 1.3.3 Condition C

[0145] Take 10 μL of each of the three solutions 1.1 and 1.2 and spot them separately on the same silica gel G thin-layer plate containing 5% silver nitrate solution. Use the lower layer solution of chloroform-ethyl acetate-methanol-water (15:40:22:10) as the developing solvent. Develop to 18 cm, remove, air dry, and spray with 0.5% potassium permanganate acetone solution for color development. Examine after color development.

[0146] 1.3.4 Existing standard for Xiegan Anshen Capsules (WS3-620(Z-103)-2004(Z))

[0147] Take 2g of the contents of Xiegan Anshen Capsules, add 20mL of ethanol, sonicate for 15 minutes, filter, discard the ethanol, evaporate the ether from the residue, add 20mL of methanol, heat under reflux in a water bath for 20 minutes, filter, evaporate the filtrate to dryness, add 15mL of water to dissolve the residue, extract twice with 20mL of water-saturated n-butanol each time, combine the n-butanol extracts, evaporate to dryness, add a small amount of anhydrous ethanol to dissolve the residue, add 1g of neutral alumina, stir well, evaporate to dryness, add to a pre-treated neutral alumina column (100-200 mesh, 2g, 1cm inner diameter), elute with 30mL of methanol, collect the eluent, evaporate to dryness, add 1mL of methanol to dissolve the residue, and use as the test solution.

[0148] Take gentiopicrin and geniposide reference standards, and prepare solutions containing 2 mg and 4 mg of each per 1 mL with methanol, respectively, as reference solutions.

[0149] Apply the above test sample and reference sample separately to the same silica gel G thin-layer plate containing 5% silver nitrate solution. Use the lower layer solution of chloroform-ethyl acetate-methanol-water (15:40:22:10) as the developing solvent. Develop to 18 cm, remove, air dry, and spray with 0.5% potassium permanganate acetone solution for color development. Examine after color development.

[0150] 2 Results Analysis

[0151] Thin-layer chromatography identification was performed using the experimental method described above, and the results are as follows:

[0152] (1) When the pharmacopoeia method under section 1.3.4 was used for detection, the spots in the chromatogram of the test sample were blurry and poorly separated at the corresponding positions as those in the chromatograms of gentiopicrin and geniposide reference standards, making them difficult to clearly identify (see Figure 1 ).

[0153] (2) When the test sample is tested under condition A in section 1.3.1, fluorescent spots of the same color appear at the corresponding positions as those of the reference standards gentiopicrin and geniposide in the chromatogram of the test sample. The spots are clear, round, and well-separated. There is no interference in the negative sample (see section 1). Figure 2 ).

[0154] (3) When the test sample is detected under condition B in section 1.3.2, only the outline of the spot is faintly visible at the corresponding position as the reference standard in the chromatogram of the test sample. The color development is blurry and it is difficult to identify effectively (see Figure 3 ).

[0155] (4) When the test sample is detected under condition C in section 1.3.3, the spots in the chromatogram of the test sample are blurred at the corresponding positions as the reference sample, with poor resolution and difficult to identify clearly (see Figure 4 ).

[0156] Example 3 Thin-layer chromatographic identification of Angelica sinensis

[0157] Preparation of test solution: Take 8g of this product, add 30mL of diethyl ether, sonicate for 15 minutes, filter, evaporate the ether from the residue (residue 1), evaporate the ether from the filtrate (filtrate 1), dissolve the residue in 1mL of ethyl acetate to obtain the test solution. Separately, take 0.5g of Angelica sinensis reference material (National Institutes for Food and Drug Control: 120927-202118), and prepare a reference material solution using the same method.

[0158] Adsorbent: Pre-fabricated silica gel G thin-layer plate. Developing solvent: n-hexane-ethyl acetate (9:1). Inspection method: Examined under ultraviolet light (365nm).

[0159] The results are as follows Figure 5 As shown, the control medicinal material shows characteristic blue-green fluorescent spots, and the test sample shows the same color spots in the corresponding positions of the control medicinal material. The negative sample shows no interference, and the spots of different batches of test samples are consistent.

[0160] Example 4: Thin-layer chromatographic identification of Alisma plantago-aquatica

[0161] Preparation of test solution: The test solution is the same as that in Example 2. Take 1g of Alisma plantago-aquatica reference material, add 40mL of ether, sonicate for 15 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1mL of ethyl acetate, and use it as the reference material solution.

[0162] Adsorbent: Glass pre-fabricated silica gel G thin-layer plate (a). Developing solvent: (I) n-hexane-ethyl acetate (1:1); (II) petroleum ether (60-90℃)-chloroform-ethyl acetate (2:1:2).

[0163] Color developer: Spray with 5% vanillin sulfuric acid solution, heat at 105℃ until the spots are clearly visible, and examine under sunlight.

[0164] The results for eluent systems I and II are as follows: Figure 6 and Figure 7 As shown, under the conditions of developing system I, the spots in the chromatogram of Alisma plantago-aquatica reference material at the corresponding positions to the 23-acetylalisenoside B reference standard are clear and of the same color, indicating that the chromatogram of the reference material contains information about the 23-acetylalisenoside B reference standard. The chromatogram of the test sample shows purplish-red spots of the same color at the corresponding positions as the reference standard and the reference material, and there are more spots corresponding to the chromatogram of the reference material. There is no interference from negative samples, and the spots of different batches of test samples are consistent. The results of developing system II are comparable to those of developing system I, but it is no longer preferred due to the involvement of precursor chemicals.

[0165] Example 5: Thin-layer chromatographic identification of Scutellaria baicalensis

[0166] Preparation of the test solution: Take the residue after ether extraction in Example 3 (filter residue 1), add 30 mL of methanol, sonicate for 15 minutes, filter, evaporate the filtrate to dryness, add 20 mL of water to dissolve the residue, add hydrochloric acid to adjust the pH to 2-3, extract twice with ethyl acetate, 20 mL each time, combine the ethyl acetate extracts, and keep the aqueous solution (aqueous solution 1). Evaporate the ethyl acetate solution to dryness, add 1 mL of methanol to dissolve the residue, and use this as the test solution. Separately, take 1 g of Scutellaria baicalensis reference material, add 40 mL of ether, sonicate for 15 minutes, filter, discard the ether solution, evaporate the ether from the residue, place it in a stoppered conical flask, add 30 mL of methanol, and prepare the reference material solution using the same method starting from "sonicate for 15 minutes, filter, evaporate the filtrate to dryness".

[0167] Adsorbent: silica gel G pre-prepared thin-layer plate containing 1% sodium hydroxide (a), silica gel G pre-prepared thin-layer plate containing 4% sodium acetate (b); Developing agent: (I) ethyl acetate-butanone-formic acid-water (5:3:1:1).

[0168] Adsorbent: Polyamide film (c); Developing agent: (II) Acetic acid. Colorimetric reagent: Sprayed with 1% ferric chloride ethanol solution and examined under sunlight.

[0169] The results of developing solvent system I on different silica gel plates are as follows: Figure 8 As shown, silica gel plate G (b) containing 4% sodium acetate exhibits spot tailing phenomenon when developed under system I conditions. Figure 8 B); Pre-prepared thin-layer plates (a) containing 1% sodium hydroxide and silica gel G showed characteristic purplish-red spots at corresponding positions of the reference medicinal material and the reference standard under System I conditions. The test sample showed spots of the same color at corresponding positions as the reference standard and the reference medicinal material. Negative samples showed no interference. Spots from different batches of test samples were consistent. Figure 8 A). System II chromatography provides more chromatographic information than System I, but the amount of polyamide film loaded is lower, and the spots are not clear enough. Figure 9 ).

[0170] Example 6: Thin-layer chromatographic identification of Polygala tenuifolia

[0171] 1 Experimental Methods

[0172] 1.1 Preparation of test solution

[0173] Take the aqueous solution 1 from Example 5, add 2 mL of hydrochloric acid, hydrolyze for 2 hours, cool, and extract twice with 20 mL of chloroform each time. Combine the chloroform extracts, evaporate to dryness, and dissolve the residue in 1 mL of methanol to obtain the test solution.

[0174] 1.2 Preparation of reference solution

[0175] Take the polygalactosine reference standard and prepare a solution containing 0.5 mg per 1 mg with methanol as the reference solution.

[0176] 1.3 Screening of Thin-Layer Chromatography Conditions

[0177] 1.3.1 Expanding System I

[0178] Using glass pre-fabricated silica gel G thin-layer plates as adsorbents, and chloroform-acetone-glacial acetic acid (9:5:0.2) as developing solvents, the plates were removed after development, dried, sprayed with 5% vanillin sulfuric acid solution, and heated at 105℃ until the spots were clearly visible. The plates were then examined under sunlight.

[0179] 1.3.2 Deployment System II

[0180] Using glass pre-fabricated silica gel G thin-layer plates as adsorbents, and chloroform-acetone-glacial acetic acid (9:1:0.2) as developing solvents, the plates were removed after development, dried, sprayed with 5% vanillin sulfuric acid solution, and heated at 105℃ until the spots were clearly visible. The plates were then examined under sunlight.

[0181] 1.3.3 Deployment System III

[0182] Using glass pre-fabricated silica gel G thin-layer plates as adsorbents, and toluene-ethyl acetate-glacial acetic acid (14:8:0.5) as developing solvents, the plates were developed, removed, dried, sprayed with 5% vanillin sulfuric acid solution, and heated at 105℃ until the spots were clearly visible. The plates were then examined under sunlight.

[0183] 1.3.4 Deployment System IV

[0184] Using glass pre-fabricated silica gel G thin-layer plates as adsorbents, and chloroform-acetone-glacial acetic acid (9:3:0.2) as developing solvents, the plates were developed, removed, dried, sprayed with 5% vanillin sulfuric acid solution, and heated at 105℃ until the spots were clearly visible. The plates were then examined under sunlight.

[0185] 2 Results Analysis

[0186] Thin-layer chromatography identification was performed using the experimental method described above, and the results are as follows:

[0187] (1) Under development system I: the reference standard shows characteristic purple spots, the test sample shows the same spots at the corresponding positions as the reference standard, the negative sample shows no interference, and the spots of different batches of test samples are consistent. Figure 10 ).

[0188] (2) Under development system II: the reference standard showed clear purple characteristic spots, and the test sample showed the same spots at the corresponding positions as the reference standard. There was no interference from the negative sample, and the spots of different batches of test samples were consistent; however, the Rf value of the polygalactosidase spots was lower ( Figure 11 ).

[0189] (3) Under development system III: the reference standard shows clear purple characteristic spots, the test sample shows the same spots at the corresponding positions as the reference standard, and the test sample and negative sample have different colored spots interfering with the spots at positions close to the reference standard spots. Figure 12 ).

[0190] (4) Under development system IV, the reference standard showed clear purple characteristic spots, and the test sample showed the same spots at the corresponding positions as the reference standard. The negative sample showed no interference, and the spots of different batches of test samples were consistent; however, the Rf value of the polygalactosidase spots was lower ( Figure 13 ).

[0191] Example 7: Simultaneous determination of gentiopicrin, geniposide, and baicalin by high performance liquid chromatography.

[0192] 1) Instruments and reagents

[0193] The equipment included an Agilent 1200 HPLC system and an Agilent 1260II HPLC system; a BS2202S electronic balance and a CPA225D electronic balance (Sartorius, Germany); a Sigma 3-30K centrifuge (Sigma, Germany); a HU20500D ultrasonic instrument (Tianjin Hengao Technology Development Co., Ltd.); methanol (chromatographic grade) purchased from Fisher Scientific; high-purity water was prepared in the laboratory; all other reagents were of analytical grade. Gentianoside (batch number: 110770-202219, purity: 98.1%), baicalin (batch number: 110715-202122, purity: 94.2%), and geniposide (batch number: 110749-201919, purity: 97.1%) were also tested.

[0194] 2) Selection of detection wavelength:

[0195] The ultraviolet absorption of gentiopicrin, geniposide, and baicalin was examined at wavelengths of 254 nm, 274 nm, 284 nm, and 317 nm. At 254 nm, all three substances showed significant absorption, exhibiting good peak height and shape. However, at wavelengths of 274 nm, 284 nm, and 371 nm, one or more of the three control substances showed poor ultraviolet absorption.

[0196] 3) Washing program selection:

[0197] Chromatographic column: Agilent ZORBAX StableBond C18 column (4.6×250mm 5μm); flow rate: 1.0mL / min; column temperature: 30℃; mobile phase: methanol (A)-0.2% phosphoric acid solution (B), eluted according to the procedures in Tables 2-4 below; detection wavelength: 254nm; injection volume: 10μL.

[0198] Table 2 Gradient elution program

[0199]

[0200] The results are as follows Figure 14 As shown, elution program 1 has the problem of interference from extraneous peaks after the geniposide peak, which affects the integration results; elution program 2 has the elution time of baicalin exceeding the range of the elution program, that is, it failed to complete the elution within the time set by the elution program; elution program 3 has appropriate elution times for gentiopicrin, geniposide, and baicalin, and there is no interference before and after the target peak.

[0201] 4) Flow rate selection

[0202] Using the elution program 3 described above, flow rates of 1.0 mL / min, 0.8 mL / min, and 1.2 mL / min were set, with a wavelength of 254 nm and an injection volume of 10 μL, which were then injected into the liquid chromatograph. At a flow rate of 0.8 mL / min, the peak elution time was generally too long, and the baicalin peak was close to the end of the gradient elution program. At a flow rate of 1.2 mL / min, the chromatographic system pressure was too high, exceeding the pressure limit of the chromatograph. At a flow rate of 1.0 mL / min, the system pressure was moderate, and the peak elution time was appropriate.

[0203] 5) Preparation of the test solution

[0204] The extraction methods were investigated using 50%, 70%, and 100% methanol as extraction solvents, and employing ultrasonic extraction (250W power, 40kHz frequency, extraction times of 15 min and 30 min) and reflux extraction (1 h, 1.5 h, and 2 h), respectively. The results are shown in Table 3. The results indicate that extraction with 70% methanol solution yielded the best results. Ultrasonic treatment for 30 minutes was comparable in efficacy to reflux extraction (1 h, 1.5 h, and 2 h), and also offered the greatest ease of operation and the shortest analysis time.

[0205] Table 3 Solvent Screening Results

[0206]

[0207] 6) Sample testing

[0208] Following the above-described content determination method, the contents of gentiopicrin, geniposide, and baicalin in 15 batches of samples were determined. The results are shown in the table below. Figure 15 The content of gentiopicroside in 15 batches of samples was determined according to the method under the existing standard (WS3-620-(Z-103)-2004Z) for content determination. The results are shown in [Figure number missing]. Figure 16 The deviation of the gentiopicroside content determined by the method of this invention from that determined by existing standards is less than 4%, indicating that the proposed method is comparable to existing standards in its ability to determine gentiopicroside content.

[0209] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A quality testing method for liver-clearing and calming capsules, characterized in that: The ingredients of the Liver-Clearing and Mind-Calming Capsules include: Gentian, Mother-of-Pearl, Scutellaria baicalensis, Gardenia jasminoides, Tribulus terrestris, Oyster shell, Dragon bone, Poria cocos, Plantago asiatica, Alisma plantago-aquatica, Ziziphus jujuba var. spinosa, Platycladus orientalis, Polygala tenuifolia, Rehmannia glutinosa, Ophiopogon japonicus, Angelica sinensis, and Glycyrrhiza uralensis. The quality testing method for the liver-clearing and calming capsules includes at least one of the following methods: (1) Thin-layer chromatography detection method for gentiopicrin and geniposide; (2) Thin-layer chromatography detection method for Scutellaria baicalensis; (3) Thin-layer chromatography detection method for Polygala tenuifolia; The thin-layer chromatography detection method for gentiopicrin and geniposide includes the following steps: Prepare test solution A of liver-clearing and calming capsules, gentiopicroside reference solution and geniposide reference solution respectively. Spot the test solution A, gentiopicroside reference solution and geniposide reference solution on a glass pre-made silica gel G thin layer plate. Develop with a mixed solvent of dichloromethane, methanol and water in a volume ratio of (25-35):(8-10):(2-4) as the developing solvent. After color development with sulfuric acid ethanol solution, perform the first inspection. The preparation of the test solution A includes the following steps: taking the contents of the liver-clearing and calming capsules and mixing them with a first organic solvent for extraction, and using the resulting extract directly as the test solution A; further, the first organic solvent is one or more of methanol, ethanol and ethyl acetate; The thin-layer chromatography detection method for Scutellaria baicalensis includes the following steps: Prepare the test solution B of Xiegan Anshen Capsules and the reference medicinal material solution of Scutellaria baicalensis. Spot the test solution B and the reference medicinal material solution of Scutellaria baicalensis on silica gel G pre-prepared thin layer plates containing 0.1%-10% sodium hydroxide. Develop the plates with a mixed solvent of ethyl acetate, butanone, formic acid and water in a volume ratio of (4-6):(2-4):1:1 as the developing solvent. After color development, perform a second inspection. The preparation of the test solution B includes the following steps: extracting the contents of the liver-clearing and calming capsules with a second organic solvent to obtain filtrate 1 and residue 1; extracting the residue 1 with a third organic solvent to obtain filtrate 2; concentrating and diluting the filtrate 2; and extracting it with ethyl acetate to obtain aqueous solution 1 and organic phase 1; concentrating and diluting the organic phase 1 to prepare the test solution B; further, the second or third organic solvent is one or more of diethyl ether, methanol, ethanol, and ethyl acetate; The thin-layer chromatography detection method for Polygala tenuifolia includes the following steps: The test solution C of Xiegan Anshen Capsules and the reference solution of Polygala tenuifolia were prepared separately. The mixture of chloroform, acetone and glacial acetic acid in a volume ratio of (8-10):(4-6):(0.1-0.3) was used as the developing solvent. After color development, the third inspection was performed. The preparation method of the test solution C includes the following steps: acidifying and hydrolyzing the aqueous solution 1, extracting it with a fourth organic solvent to obtain an organic phase 2, concentrating and diluting the organic phase 2, and preparing the test solution C; further, the fourth organic solvent is one or more of chloroform, dichloromethane, carbon tetrachloride, and ethyl acetate.

2. The method for detecting the liver-clearing and calming capsules according to claim 1, characterized in that: In the thin-layer chromatography method for detecting gentiopicrin and geniposide, the concentration of the sulfuric acid ethanol solution is 5%-15%; further, the concentration of the sulfuric acid ethanol solution is 10%.

3. The method for detecting the liver-clearing and calming capsules according to claim 1, characterized in that: In the thin-layer chromatography detection method for Scutellaria baicalensis, the thin-layer plate is a silica gel G pre-prepared thin-layer plate containing 0.5%-5% sodium hydroxide; further, the thin-layer plate is a silica gel G pre-prepared thin-layer plate containing 1% sodium hydroxide.

4. The method for detecting the liver-clearing and calming capsules according to claim 1, characterized in that: In the thin-layer chromatography detection method for Polygala tenuifolia, the volume ratio of chloroform, acetone and glacial acetic acid is 9:5:0.

2.

5. The quality testing method for the liver-clearing and calming capsules according to claim 1, characterized in that: The preparation method of the gentiopicroside reference solution and the geniposide reference solution includes the following steps: taking gentiopicroside reference standard and geniposide reference standard respectively, dissolving them in a polar organic solvent to obtain the gentiopicroside reference solution and the geniposide reference solution; further, the polar organic solvent is one or more of methanol, ethanol and ethyl acetate; The preparation method of the Scutellaria baicalensis reference herb solution includes the following steps: Scutellaria baicalensis reference herb is mixed with diethyl ether for extraction to obtain filter residue 2; the filter residue 2 is mixed with methanol for extraction to obtain filtrate 3; the filtrate 3 is concentrated, diluted, and then mixed with ethyl acetate for extraction to obtain organic phase 3; the organic phase is concentrated, diluted, and the Scutellaria baicalensis reference herb solution is prepared. The preparation method of the Polygala tenuifolia reference solution includes the following steps: taking Polygala tenuifolia saponin reference standard, dissolving it in a polar organic solvent to obtain the Polygala tenuifolia reference solution; further, the polar organic solvent is one or more of methanol, ethanol and ethyl acetate.

6. The quality testing method for the liver-clearing and calming capsules according to claim 1, characterized in that: The detection methods also include thin-layer chromatography detection methods for Angelica sinensis and Alisma plantago-aquatica. The thin-layer chromatography detection method for Angelica sinensis includes the following steps: preparing the test solution D of Xiegan Anshen Capsules and the reference medicinal material solution of Angelica sinensis respectively; spotting the test solution D and the reference medicinal material solution of Angelica sinensis on a glass prefabricated silica gel G thin-layer plate; developing the plate with a mixed solvent of n-hexane and ethyl acetate in a volume ratio of (8-10):1 as the developing solvent; and performing a fourth inspection. The thin-layer chromatography detection method for Alisma plantago-aquatica includes the following steps: developing the test solution E of Xiegan Anshen Capsules and the reference herb solution of Alisma plantago-aquatica with a volume ratio of 1:1 using a mixed solvent of n-hexane and ethyl acetate as the developing solvent, and performing a fifth inspection after color development with a colorimetric reagent.

7. The quality testing method for the liver-clearing and calming capsules according to claim 6, characterized in that: The preparation of the test solution D includes the following steps: concentrating and diluting the filtrate 1 to prepare the test solution D; The preparation of the Angelica sinensis reference herb solution includes the following steps: mixing the Angelica sinensis reference herb with a second organic solvent for extraction to obtain filtrate 4, concentrating and diluting the filtrate 4, and preparing the Angelica sinensis reference herb solution; The preparation of the test solution E includes the following steps: taking the test solution A as the test solution E; The preparation of the Alisma plantago-aquatica reference herb solution includes the following steps: Alisma plantago-aquatica reference herb is mixed with ether for extraction to obtain filtrate 5, and the filtrate 5 is concentrated, diluted, and prepared into the Alisma plantago-aquatica reference herb solution.

8. The quality testing method for the liver-clearing and calming capsules according to claim 1, characterized in that: The detection method further includes a high-performance liquid chromatography method for simultaneously determining the contents of gentiopicrin, geniposide, and baicalin, comprising the following steps: (1) Sample pretreatment: Mix the contents of Xiegan Anshen capsules with 50-100% methanol solution to extract the sample solution to be tested; preferably 70% methanol solution; (2) Chromatographic analysis: The sample solution to be tested was analyzed and determined by high performance liquid chromatography; The elution process used octadecylsilane-bonded silica gel as the packing material; methanol as mobile phase A and 0.2% phosphoric acid solution as mobile phase B; the gradient elution program was as follows: 。

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