阅读说明：本技术 一种杨树花药材的梯度多信息薄层鉴别方法 (Gradient multi-information thin-layer identification method for poplar flower medicinal materials ) 是由 缪雨臻 兰新财 缪雪荣 安丽娜 韩桂茹 麻军法 赵丽丽 于 2019-08-28 设计创作，主要内容包括：本发明涉及一种杨树花药材的梯度多信息薄层鉴别方法。其特征在于：用甲醇超声,上清液直接作为供试品与对照药材溶液；采用同该供试品溶液,以5种不同极性的展开剂,在5块普通的薄层板上,多种检视条件下,检视了杨树花药材不同极性成分的斑点30余个。各成分相互交叉,但在各自的检视条件下,互不干扰,均能呈现出清晰的颜色斑点或荧光斑点。完成该薄层鉴别只需药材和对照药材各0.3g、提取溶剂4ml、展开剂50ml、时间1.5小时。首次提供了一套杨树花药材从脂溶性到水溶性梯度多信息薄层鉴别图谱和鉴别方法。方法简便、快捷、高效,为不同工艺制备的杨树花制剂,提供了薄层参考,提升了杨树花质量评价监督指标。(The invention relates to a gradient multi-information thin-layer identification method for poplar flower medicinal materials. The method is characterized in that: performing ultrasonic treatment with methanol, and collecting supernatant as test solution and control solution; more than 30 spots of poplar flower medicinal materials with different polarity components are inspected on 5 common thin-layer plates by using 5 developing agents with different polarities and the same test solution under various inspection conditions. The components are crossed with each other, but do not interfere with each other under the respective inspection conditions, and clear color spots or fluorescent spots can be displayed. The thin layer identification is completed by only 0.3g of each of the medicinal material and the reference medicinal material, 4ml of extraction solvent and 50ml of developing solvent for 1.5 hours. Provides a set of thin-layer multilayer information identification maps and identification methods for medicinal materials of poplar flowers from fat solubility to water solubility gradient for the first time. The method is simple, convenient, rapid and efficient, provides a thin-layer reference for poplar flower preparations prepared by different processes, and improves the quality evaluation and supervision indexes of poplar flowers.)

1. A gradient multi-information thin-layer identification method for poplar flower medicinal materials is characterized by comprising the following steps: the method comprises the following steps: (1) the method comprises the following steps of (1) carrying out thin-layer identification on fat-soluble components of a poplar flower medicinal material, (2) carrying out thin-layer identification on partial fat-soluble components of the poplar flower medicinal material, (3) carrying out thin-layer identification on polar components in the poplar flower medicinal material, (4) carrying out thin-layer identification on partial water-soluble components of the poplar flower medicinal material, and (5) carrying out thin-layer identification on water-soluble components of the poplar flower medicinal material;

(1) performing thin-layer identification on fat-soluble components of poplar flower medicinal material, grinding 0.3g of poplar flower medicinal material and a reference medicinal material respectively, adding 2ml of methanol respectively, performing ultrasonic treatment for 20 minutes, and taking supernate as a test sample and poplar flower reference medicinal material solution; respectively dripping 5-6 μ l of reference medicinal material solution and sample solution on the same silica gel GF₂₅₄Spreading on a thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 20: 1: 0.1, taking out, drying with hot air, spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, inspecting under ultraviolet lamp 365nm, and displaying fluorescent main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the control medicinal material; inspecting in sunlight, wherein main spots of the same color appear in the chromatogram of the test solution at the positions corresponding to those in the chromatogram of the reference solution;

(2) Thin-layer identification of fat-soluble components of poplar flower medicinal material, namely, respectively dropping 5-6 mul of test solution and reference medicinal material solution under the item (1) on the same silica gel GF₂₅₄Spreading on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 8: 2: 0.1, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; inspecting under 254nm ultraviolet lamp; in the chromatogram of the test solution, main spots of the same color appear at the corresponding positions of the chromatogram of the reference solution; spraying 10% ethanol sulfate solution, and heating at 105 deg.C to spotThe color is clear, the sample is inspected under 365nm ultraviolet lamp, and a plurality of fluorescent spots with the same color are displayed on the corresponding positions of the chromatogram of the reference drug in the chromatogram of the sample; then placing the sample in a darkroom, and displaying a plurality of spots with the same color in the chromatogram of the sample at the positions corresponding to the chromatograms of the reference medicinal materials through light inspection;

(3) Thin-layer identification of polar components in poplar flower medicinal material 5-6 mul of each of the test solution and the reference medicinal material solution under item (1) are respectively spotted on the same silica gel GF₂₅₄Spreading on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 2: 4: 0.1, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; inspecting under 254nm ultraviolet lamp; in the chromatogram of the test solution, multiple brown spots appear at the positions corresponding to those of the chromatogram of the control solution; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color clearly, and inspecting under ultraviolet lamp 365nm to obtain multiple fluorescent spots with the same color in the sample chromatogram and the corresponding position of the control medicinal material chromatogram; then placing the sample in a darkroom, and displaying a plurality of spots with the same color in the chromatogram of the sample at the positions corresponding to the chromatograms of the reference medicinal materials through light inspection;

(4) thin-layer identification of water-soluble components of poplar flower medicinal material, absorbing 5-6 mul of each of the test solution and the reference medicinal material solution under the item (1), and respectively dropping the same silica gel GF₂₅₄Spreading chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent at volume ratio of 2: 3: 4: 0.5 on the thin layer plate, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; inspecting under 254nm ultraviolet lamp; displaying a plurality of spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clear, and inspecting under 365nm ultraviolet lamp; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; then the sample is placed in a dark room and inspected by lamplight to be in the chromatogram of the sampleDisplaying a plurality of spots with the same color at the corresponding positions of the chromatogram of the reference medicinal material;

(5) Thin-layer identification of water-soluble components of poplar flower medicinal material 5-6 mul of each of the test solution and the reference medicinal material solution under item (1) is respectively spotted on the same silica gel GF₂₅₄spreading chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent at volume ratio of 1: 2: 4.5: 0.5 on the thin layer plate, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; inspecting under 254nm ultraviolet lamp; at least 2 spots with the same color appear on the chromatogram of the test solution at the positions corresponding to the chromatogram of the reference solution; spraying 10% ethanol sulfate solution, heating at 105 deg.C until yellow spots appear, and inspecting under 365nm ultraviolet lamp; displaying 1 same blue-green fluorescence main spot on the corresponding position of the chromatogram of the test solution and the chromatogram of the reference solution; heating until the spots are clear, and inspecting in dark room with lamp light to show main spots of the same color in the chromatogram of the sample at the positions corresponding to those in the chromatogram of the reference medicinal material.

2. The gradient multi-information thin-layer identification method of poplar flower medicinal materials according to claim 1, wherein about 30 characteristic spots are detected on 5 thin-layer plates by 5 developing agents with different polarities according to the arrangement sequence of non-polarity, medium polarity and polarity, the total time is 1.5 hours, the extraction solvent is 4ml and the developing agent is 50 ml.

Technical Field

the invention relates to a gradient multi-information thin-layer identification method for poplar flower medicinal materials. That is, more than 30 information spots of the poplar flower medicinal material are inspected on 5 thin-layer plates by using 5 developing agents with different polarities under various inspection conditions. The thin layer identification is completed by only 0.3g of each of the medicinal material and the reference medicinal material, 4ml of extraction solvent and 50ml of developing solvent for 1.5 hours.

Background

The gradient complete information thin-layer identification of the traditional Chinese medicinal materials is characterized in that the gradient complete information thin-layer identification of the high-performance liquid phase is extended to the field of the thin-layer identification of the traditional Chinese medicinal materials, fat-soluble, medium-polar and water-soluble components in the traditional Chinese medicinal materials are repeatedly adsorbed and desorbed in developing solvents with different polarities according to the similar compatible dissolving law and the sequence of non-polarity, medium-polarity and polarity, most effective components with detection information in the medicinal materials are obtained on a few thin-layer plates according to the polarity size under different conditions, and a spot information map which is as complete as possible is obtained, so that a set of complete information thin-layer chromatogram and identification methods from the fat-soluble to the water-soluble components are formed, and various preparations prepared by different processes are provided to serve as detection indexes for quality evaluation; or providing poplar flower medicinal materials with different sources and different harvesting seasons for overall quality evaluation and supervision.

The Chinese medicine poplar flower is a variety collected in Chinese pharmacopoeia, and is dry male inflorescence of poplar, Canada poplar or other plants in the same genus. Has the effects of clearing away heat and toxic materials, eliminating dampness and relieving diarrhea, and can be used for treating damp-heat dysentery and diarrhea of young livestock. Mainly contains flavone, organic acid, amino acid, phenols, saccharides, alkaloid, etc. The flavonoids in the poplar flower extract can effectively inhibit the synthesis of pathogen nucleic acid, thereby achieving the purpose of quickly killing pathogens, having obvious inhibiting and killing effects on dysentery bacillus, escherichia coli, salmonella, proteus, staphylococcus, pasteurella, pseudomonas aeruginosa and haemophilus parasuis, having the total effective rate of more than 95 percent and having better curative effect than florfenicol and cephalosporins. Is a main medicinal material for treating dysentery and enteritis for common animals. The chemical components of the composition are fat-soluble, medium-polar and water-soluble. The method for identifying the poplar flower item in Chinese veterinary pharmacopoeia by using a thin layer comprises the following steps:

Collecting poplar flower 1g, adding water 15ml, heating and refluxing for 1 hr, filtering, extracting the filtrate with ethyl acetate for 2 times (10 ml each time), discarding ethyl acetate layer, collecting the lower layer solution, evaporating to dryness, and dissolving the residue with methanol 2ml to obtain sample solution. Preparing another 1g of poplar flower reference medicinal material, and preparing a reference medicinal material solution by the same method. Testing by thin layer chromatography (general rule 0502), sucking 5 μ l of the above two solutions, respectively dropping on the same silica gel H thin layer plate, developing with ethyl acetate-acetone-methanol-water (6.5: 4: 0.8) as developing agent, taking out, air drying, spraying with aluminum trichloride test solution, heating at 105 deg.C for 1 min, and inspecting under ultraviolet lamp (365 nm). The main fluorescent spots with the same color appear on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution. Therefore, the method has certain complexity, the time for detecting a batch of samples is 2.5 hours, the organic solvent is 44ml, the developing agent is 10ml, and only the main spots of the water-soluble flavonoids are detected.

A plurality of documents are consulted, the existing methods of collecting and carrying either oral liquid or tablets are basically carried out by adopting a veterinary drug dictionary of 2015 edition, the detected flavonoid is the water-soluble flavonoid, and other methods are not consulted. The detection index is deficient, the method is single, and the overall quality evaluation and supervision of the medicinal materials are not facilitated.

In order to improve the current situation of the current thin-layer identification of the poplar flower medicinal material, a simple, convenient, popular and multi-information gradient thin-layer identification method is provided, and the special thin-layer identification research is carried out on the poplar flower medicinal material. A set of simple, convenient and quick gradient multi-information thin-layer identification method and a map without environmental pollution are obtained.

Disclosure of Invention

A test sample and a reference medicinal material solution are obtained by a simple and quick pretreatment method, and more than 30 information spots with different polarities of the poplar flower medicinal material are inspected on 5 common thin-layer plates by using the same test sample solution and 5 developing agents with different polarities under various inspection conditions. The components are crossed with each other, but do not interfere with each other under the respective inspection conditions, and clear color spots or fluorescent spots can be displayed. The thin layer identification is completed by only 0.3g of each of the medicinal material and the reference medicinal material, 4ml of extraction solvent and 50ml of developing solvent for 1.5 hours. Provides a set of thin-layer multilayer information identification maps and identification methods for medicinal materials of poplar flowers from fat solubility to water solubility gradient for the first time. The method is simple, convenient, rapid and efficient, provides a thin layer reference for poplar flower preparations prepared by different processes, and improves the quality supervision information index of poplar flowers.

The technical scheme adopted by the invention for solving the technical problems is as follows:

(1) Performing thin-layer identification on fat-soluble components of poplar flower medicinal material, grinding 0.3g of poplar flower medicinal material and a reference medicinal material respectively, adding 2ml of methanol respectively, performing ultrasonic treatment for 20 minutes, and taking supernate as a test sample and poplar flower reference medicinal material solution; respectively dripping 5-6 μ l of reference medicinal material solution and sample solution on the same silica gel GF₂₅₄Spreading on a thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 20: 1: 0.1, taking out, drying with hot air, spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, inspecting under ultraviolet lamp 365nm, and displaying fluorescent main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the control medicinal material; and (4) observing under sunlight, wherein main spots with the same color appear in the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference medicinal materials.

(2) Thin-layer identification of fat-soluble components of poplar flower medicinal material, namely, respectively dropping 5-6 mul of test solution and reference medicinal material solution under the item (1) on the same silica gel GF₂₅₄Spreading on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 8: 2: 0.1, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; inspecting under 254nm ultraviolet lamp; in the chromatogram of the test solution, main spots of the same color appear at the corresponding positions of the chromatogram of the reference solution; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, inspecting under ultraviolet lamp 365nm, testing in the chromatogram of the sample,displaying a plurality of fluorescent spots with the same color at the positions corresponding to the color spectrums of the reference medicinal materials; then the sample is placed in a darkroom and inspected through lamplight, and a plurality of spots with the same color are displayed on the corresponding positions of the chromatogram of the test drug and the chromatogram of the reference drug.

(3) Thin-layer identification of polar components in poplar flower medicinal material 5-6 mul of each of the test solution and the reference medicinal material solution under item (1) are respectively spotted on the same silica gel GF₂₅₄Spreading on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 2: 4: 0.1, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; inspecting under 254nm ultraviolet lamp; in the chromatogram of the test solution, multiple brown spots appear at the positions corresponding to those of the chromatogram of the control solution; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color clearly, and inspecting under ultraviolet lamp 365nm to obtain multiple fluorescent spots with the same color in the sample chromatogram and the corresponding position of the control medicinal material chromatogram; then the sample is placed in a darkroom and inspected through lamplight, and a plurality of spots with the same color are displayed on the corresponding positions of the chromatogram of the test drug and the chromatogram of the reference drug.

(4) Thin-layer identification of water-soluble components of poplar flower medicinal material by respectively taking 5-6 mul of test solution and reference medicinal material solution under the item (1) and respectively dropping the test solution and the reference medicinal material solution on the same silica gel GF₂₅₄Spreading chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent at volume ratio of 2: 3: 4: 0.5 on the thin layer plate, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; inspecting under 254nm ultraviolet lamp; displaying a plurality of spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clear, and inspecting under 365nm ultraviolet lamp; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; then placing in a darkroom, observing through lamplight, wherein multiple samples in the chromatogram of the sample are the same at the corresponding positions of the chromatogram of the reference medicinal materialSpots of color.

(5) Thin-layer identification of water-soluble components of poplar flower medicinal material 5-6 mul of each of the test solution and the reference medicinal material solution under item (1) is respectively spotted on the same silica gel GF₂₅₄Spreading chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent at volume ratio of 1: 2: 4.5: 0.5 on the thin layer plate, taking out, drying with hot air, and inspecting under ultraviolet lamp 365 nm; displaying a plurality of fluorescent spots with the same color on the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution; inspecting under 254nm ultraviolet lamp; at least 2 spots with the same color appear on the chromatogram of the test solution at the positions corresponding to the chromatogram of the reference solution; spraying 10% ethanol sulfate solution, heating at 105 deg.C until yellow spots appear, and inspecting under 365nm ultraviolet lamp; displaying 1 same main fluorescent spot of brilliant blue green at the position corresponding to the chromatogram of the reference medicinal material in the chromatogram of the test solution; heating until the spots are clear, and inspecting in dark room with lamp light to show main spots of the same color in the chromatogram of the sample at the positions corresponding to those in the chromatogram of the reference medicinal material.

the principle of the invention is as follows:

According to the chemical structure and properties of each effective component of the traditional Chinese medicine, a test sample and a reference medicinal solution are simply, conveniently and quickly prepared by adopting a proper extraction solvent according to a similar compatible extraction principle. And then, the developing agents with different polarities are adopted for development, and various chemical components can be well separated on respective thin-layer plates along with different developing agents according to different adsorption, desorption, re-adsorption and re-desorption capacities. And then, by means of effective components with similar polarities, the effective components are overlapped on the same thin-layer plate under different inspection conditions, but do not interfere with each other on different layers, so that different spot colors are presented, and a thin-layer chromatogram with multiple information is obtained. And according to the polarity of the developing agent, well-separated multi-information gradient thin-layer chromatograms on each thin-layer plate are connected from non-polarity to form a set of holographic thin-layer chromatograms.

The invention has the following innovation points and beneficial effects:

1. The poplar flower medicinal material takes cyclohexane-ethyl acetate-formic acid with the volume ratio of 20: 1: 0.1 as a developing agent, after the developing, no spot information exists under the ultraviolet light 365nm and 254nm, but 1 bright light blue fluorescence main spot and 2 blue spots with lower Rf value are detected under the ultraviolet light 365nm after the spraying of 10% sulfuric acid ethanol solution for color development (figure 1), the main spot is pale pink under the sunlight, and the 2 fluorescence spots with lower Rf value are not clear (figure 2). Therefore, only the main spots were observed among the fat-soluble components.

2. The poplar flower medicinal material is developed by taking cyclohexane-ethyl acetate-formic acid with the volume ratio of 8: 2: 0.1 as a developing agent, and 4 fluorescent spots, one red, one green and two blue spots are detected under an ultraviolet lamp at 365nm (figure 3); 2 tan spots observed under 254nm UV lamp, which are not in the same position and not in the same composition as the fluorescent spots from Rf value (FIG. 4); spraying 10% sulfuric acid ethanol solution for color development, inspecting 6 spots under an ultraviolet lamp at 365nm, wherein 2 light pink fluorescent spots, 2 light yellow green fluorescent spots, 1 blue fluorescent spot and 1 tan non-fluorescent spot (figure 5) are arranged in a dark room, and 5 spots with different colors are displayed through light inspection (figure 6); on the same thin-layer plate, 10 fat-soluble component spots were detected under 4 different inspection conditions.

3. Taking cyclohexane-ethyl acetate-formic acid with a volume ratio of 2: 4: 0.1 as a developing agent, and inspecting 3 fluorescent spots with different colors under an ultraviolet lamp at 365nm after the developing, wherein 2 fluorescent spots are yellowish green and one is bluish-purple (figure 7); 2 tan spots observed under 254nm UV lamp were not in the same position and not in the same composition as the fluorescent spot in Rf value (FIG. 8); spraying 10% ethanol sulfate solution to develop color, and inspecting 3 fluorescent spots under ultraviolet lamp 365nm, wherein 2 fluorescent spots are yellowish green and 1 fluorescent blue (FIG. 9); then the film is placed in a darkroom and is inspected through lamplight, 3 color spots (figure 10) are presented, and the Rf value shows that the color spots are not the same as the developed fluorescent spots; on the same thin-layer plate, 9 medium-polarity component spots were detected under 4 different inspection conditions.

4. Developing the poplar flower medicinal material with chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent at volume ratio of 2: 3: 4: 0.5, and inspecting under ultraviolet lamp 365nm to obtain 4 fluorescent spots, 1 green and 3 brilliant blue-purple (FIG. 11); 4 tan spots were observed under UV 254nm, 3 of which, from the Rf value, were not in the same position and not of the same composition as the fluorescent spot (FIG. 12); spraying 10% ethanol sulfate solution to develop color, and inspecting 5 fluorescent spots under ultraviolet lamp 365nm, wherein 4 fluorescent spots are yellow green and 1 fluorescent spot is bright blue (FIG. 13); then the film is placed in a darkroom and is inspected through lamplight, 6 color spots (figure 14) are shown, and from the Rf value, 3 of the color spots and the developed fluorescence spots are not the same components; on the same thin layer plate, 9 spots of water-soluble components were detected under 4 different inspection conditions.

5. The poplar flower medicinal material is developed by using chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent in the volume ratio of 1: 2: 4.5: 0.5, and 3 fluorescent spots, 1 green and 2 brilliant blue-purple spots are detected under the ultraviolet lamp of 365nm (figure 15); 2 tan spots were observed under a 254nm UV lamp, which were not in the same position and not in the same composition as the fluorescent spots from the Rf value (FIG. 16); spraying 10% ethanol sulfate solution to develop color, and inspecting 1 very bright blue-green main fluorescent spot under 365nm ultraviolet lamp (FIG. 17); then the sample is placed in a dark room and inspected by lamplight, so that 1 tan spot (figure 18) appears, and the Rf value shows that the spot is not the same as the fluorescent spot; on the same thin layer plate, 5 water-soluble components are detected under 4 different inspection conditions.

6. Using 5 developing agents with different polarities, and detecting 34 fat-soluble, medium-polar and water-soluble information spots which are not coincident with each other on 5 thin-layer plates under 4 inspection conditions to form a set of poplar flower complete information gradient thin-layer identification method and provide a reference map. The research of the system has not been reported, and the system has novelty, novelty and practicability.

7. The method is simple, convenient and rapid, and only methanol ultrasound is needed for medicinal materials and reference medicinal materials, and the sample application and the development of the supernatant are needed. The color developing agent is a very common 10% sulfuric acid ethanol solution, is a common color developing agent, can react with most organic compounds to present fluorescent spots or spots with different colors, is cheap and easy to obtain, and is beneficial to popularization and promotion of the method.

8. The identification method only needs 0.3g of each of the medicinal materials and the reference medicinal materials, 4ml of extraction solvent, 50ml of developing solvent and 1.5 hours. Compared with the veterinary drug classical method, the method can save 1 hour for detecting a batch of medicinal materials, but the detected information spots are at least 10 times of those of the veterinary pharmacopoeia. Compared with the information spots of a single thin-layer plate, the information spots of the traditional Chinese medicine preparation are more than those of a veterinary medicine preparation.