Thin-layer chromatography method for simultaneously identifying rhizoma coptidis and cortex phellodendri components and application of thin-layer chromatography method

Abstract

The invention relates to the technical field of traditional Chinese medicine detection, and particularly discloses a thin-layer chromatography method for simultaneously identifying coptis chinensis and golden cypress components and application of the thin-layer chromatography method. The invention provides a thin-layer chromatography method for simultaneously identifying components of coptis chinensis and golden cypress. The thin-layer chromatography method comprises the following steps: (1) respectively preparing a test solution, a coptis chinensis control medicinal material solution, a golden cypress control medicinal material solution and a control solution; and (2) respectively dripping the solutions on the same silica gel G thin-layer plate, putting the silica gel G thin-layer plate into a developing cylinder by using normal hexane, normal butanol, methanol and water in a volume ratio of (2.5-3.5): (2.5-3.5): (1.5-2.5): (0.8-1.2) as a developing solvent, developing, taking out, airing, and inspecting under a light source of 360-370nm. The thin-layer chromatography method provided by the invention can be used for quickly, simply and conveniently identifying the coptis chinensis and cortex phellodendri components in the traditional Chinese medicine external preparation, so that the quality control of the traditional Chinese medicine external preparation is realized.

Description

Technical Field

[0001] The present application relates to the technical field of traditional Chinese medicine detection, and in particular to a thin layer chromatography method for simultaneously identifying components of coptis chinensis and phellodendron amurense and its application. Background Art

[0002] With the acceleration of the modernization of traditional Chinese medicine, the effectiveness and safety of Chinese medicine preparations have received more and more attention. Among them, ingredient identification is one of the key steps to ensure the quality of drugs, and its importance is self-evident. Although modern instrumental analysis methods such as high-performance liquid chromatography (HPLC) and gas chromatography (GC) can quickly realize drug quality detection, they are costly and complex to operate, making them difficult to be widely used in daily quality control work. Thin-layer chromatography research is widely used in the identification of effective ingredients in traditional Chinese medicines because of its low cost and fast response time.

[0003] At present, there are related documents reporting a thin layer chromatography detection method for the simultaneous detection of coptis chinensis and phellodendron amurense in preparations. This method uses the "secondary thin layer development method" to achieve the identification of coptis chinensis and phellodendron amurense components in the Danggui Liuhuang Decoction compound preparation. However, this method uses two developing reagents for two developments, and the inspection after the second development also requires the use of a color developer for color development. Therefore, this method is cumbersome to operate, has lengthy steps, and has a high time cost.

[0004] Therefore, there is an urgent need to improve the existing thin layer chromatography method, so as to establish a thin layer chromatography method that can quickly and easily identify the two active ingredients of Coptis chinensis and Phellodendron chinense in traditional Chinese medicine preparations at the same time, so as to meet the daily quality control work of traditional Chinese medicine preparations. Summary of the invention

[0005] In order to quickly and easily identify the two effective ingredients of Coptis chinensis and Phellodendron amurense in traditional Chinese medicine preparations at the same time, the present application provides a thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron amurense and its application.

[0006] In the first aspect, the present application provides a thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense, using the following technical solution: A thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense comprises the following steps: (1) Prepare the test sample solution, Rhizoma Coptidis reference medicinal material solution, Rhizoma Phellodendri reference medicinal material solution and reference substance solution respectively; (2) Spot the above solutions on the same silica gel G thin layer plate respectively, use n-hexane-n-butanol-methanol-water with a volume ratio of (2.5-3.5):(2.5-3.5):(1.5-2.5):(0.8-1.2) as the developing agent, place the silica gel G thin layer plate in a developing tank for development, and examine it under a light source of 360-370nm.

[0007] Optionally, the volume ratio of n-hexane-n-butanol-methanol-water is 3:3:2:1.

[0008] Optionally, the spotting volume of each solution is 3-8 μL.

[0009] Optionally, the development cylinder is pre-saturated with concentrated ammonia solution for 20-30 minutes.

[0010] Optionally, the test solution is prepared by adding water to the sample to be tested, heating to melt, then heating under reflux and stirring at 400-450 rpm for 30-40 min, and cooling to room temperature; then storing at 0±5°C for 25-35 min, filtering, extracting, evaporating the aqueous phase, and adding methanol to dissolve to obtain the test solution.

[0011] Optionally, the sample to be tested is Coptis chinensis paste.

[0012] Optionally, the preparation method of the Coptis chinensis control medicinal material solution is: add water to the Coptis chinensis control medicinal material, then heat and reflux with stirring at 400-450 rpm for 30-40 minutes, and cool to room temperature; filter, extract, evaporate the water phase, and add methanol to dissolve to obtain the Coptis chinensis control medicinal material solution.

[0013] Optionally, the preparation method of the Phellodendron chinense reference medicinal material solution is: add water to the Phellodendron chinense reference medicinal material, then heat and reflux with stirring at 400-450rpm for 30-40min, and cool to room temperature; filter, extract, evaporate the water phase, and add methanol to dissolve to obtain the Phellodendron chinense reference medicinal material solution.

[0014] Optionally, the reference solution is prepared by taking a berberine hydrochloride reference substance, adding methanol to dissolve it, and obtaining a reference solution with a concentration of 0.05 mg / mL.

[0015] In a second aspect, the present application provides a thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense for use in identifying the components of Coptis chinensis and Phellodendron chinense in external Chinese medicine preparations.

[0016] Optionally, the Chinese medicine external preparation is Coptis chinensis paste.

[0017] In the present application, coptis paste is a common external preparation of traditional Chinese medicine, the main ingredients of which usually include coptis root, phellodendron, etc., and it has a long history and wide clinical application. The thin layer chromatography method provided in the present application can quickly and easily identify the coptis root and phellodendron components in coptis paste, realize the quality control of coptis paste, has the advantages of high accuracy and short identification time, and is suitable for large-scale promotion and use.

[0018] In summary, this application has the following beneficial effects: The present application provides a thin layer chromatography method for simultaneously identifying the components of coptis chinensis and phellodendron chinense. The detection method uses n-hexane-n-butanol-methanol-water with a volume ratio of (2.5-3.5): (2.5-3.5): (1.5-2.5): (0.8-1.2) as a developing solvent. After one development, the coptis chinensis and phellodendron chinense components in traditional Chinese medicine external preparations can be qualitatively identified. Compared with the thin layer chromatography detection method in the related art, the above method has the advantages of low solvent cost, simple operation, short identification time, etc., which can meet the daily work of identifying the effective ingredients in traditional Chinese medicine external preparations and realize the quality control of traditional Chinese medicine external preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a thin layer chromatogram of the specificity investigation result; from left to right: 1 is berberine hydrochloride reference solution; 2 is coptis chinensis reference medicinal material solution; 3 is phellodendron chinense reference medicinal material solution; 4 is HLG-1002-1073 coptis chinensis paste test solution; 5 is HLG-1002-1075 coptis chinensis paste negative lack of coptis chinensis test solution; 6 is HLG-1002-1076 coptis chinensis paste negative lack of phellodendron chinense test solution; 7 is HLG-1002-1078 coptis chinensis paste double negative lack of coptis chinensis and phellodendron chinense test solution; Figure 2 It is a thin layer chromatogram obtained by using silica gel G thin layer plates produced by different manufacturers; Figure 2 In the figure, (a) is the German Merck manufacturer, (b) is the Yantai Huayang manufacturer, and (c) is the Yantai Jiangyou manufacturer; from left to right in each chromatogram are: 1 is berberine hydrochloride reference solution; 2 is coptis chinensis reference medicinal material solution; 3 is phellodendron chinense reference medicinal material solution; 4 is HLG-1002-1166 coptis chinensis paste test solution 1; 5 is HLG-1002-1167 coptis chinensis paste test solution 2; Figure 3 Thin layer chromatograms obtained for silica gel G thin layer plate detection under different environmental conditions; Figure 3 Among them, (a) is Qingdao Ocean manufacturer, 4℃ low temperature, 67% humidity, (b) is Qingdao Ocean manufacturer, 25℃, 92% humidity, (c) is Yantai Jiangyou manufacturer, 4℃ low temperature, 67% humidity, (d) is Yantai Jiangyou manufacturer, 25℃, 92% humidity; from left to right in each chromatogram: 1 is berberine hydrochloride reference solution; 2 is Coptis chinensis reference medicinal material solution; 3 is Phellodendron chinense reference medicinal material solution; 4 is HLG-1002-1073 Coptis chinensis paste test solution; 5 is HLG-1002-1075 Coptis chinensis paste negative lack of Coptis chinensis test solution; 6 is HLG-1002-1076 Coptis chinensis paste negative lack of Phellodendron chinense test solution; 7 is HLG-1002-1078 Coptis chinensis paste double negative lack of Coptis chinensis and Phellodendron chinense test solution. DETAILED DESCRIPTION

[0020] The present application provides a thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense, comprising the following steps: (1) Prepare the test sample solution, Rhizoma Coptidis reference medicinal material solution, Rhizoma Phellodendri reference medicinal material solution and reference substance solution respectively; (1-1) Test solution: add water to the sample to be tested, heat to melt, then heat under reflux at 400-450 rpm with stirring for 30-40 min, cool to room temperature; then store at 0±5°C for 25-35 min, filter, extract, evaporate the aqueous phase, and add methanol to dissolve to obtain a test solution; (1-2) Coptis chinensis reference medicinal material solution: The method for preparing the Coptis chinensis reference medicinal material solution is as follows: water is added to the Coptis chinensis reference medicinal material, and then heated under reflux and stirred at 400-450 rpm for 30-40 minutes, and cooled to room temperature; filtering, extracting, evaporating the aqueous phase, and adding methanol to dissolve to obtain the Coptis chinensis reference medicinal material solution; (1-3) Phellodendron chinense reference medicinal material solution: The preparation method of the Phellodendron chinense reference medicinal material solution is as follows: water is added to the Phellodendron chinense reference medicinal material, and then heated under reflux and stirred at 400-450 rpm for 30-40 minutes, and then cooled to room temperature; after filtering, extracting, evaporating the water phase, and adding methanol to dissolve, a Phellodendron chinense reference medicinal material solution is obtained; (1-4) Take berberine hydrochloride reference substance, add methanol to dissolve, and obtain a reference substance solution with a concentration of 0.05 mg / mL; (2) Spot the above solutions on the same silica gel G thin layer plate, and use n-hexane-n-butanol-methanol-water with a volume ratio of (2.5-3.5):(2.5-3.5):(1.5-2.5):(0.8-1.2) as the developing agent. Place the silica gel G thin layer plate in a developing cylinder that has been saturated with concentrated ammonia solution for 20-30 minutes, develop, take out, dry, and then examine under a light source of 360-370nm.

[0021] In this application, the prescription of Coptis chinensis paste is 20g Coptis chinensis, 20g Phellodendron chinense, 20g Curcuma longa, 33g Angelica sinensis, and 66g Rehmannia glutinosa. The preparation method is: mix the above 5 flavors, weigh 792g sesame oil, heat to 180-190℃, add the mixed pieces and extract for 15 minutes, filter, weigh 264g beeswax, filter after melting, stir with the oil, cool, and make 1000g. The batch number of the coptis paste used in the examples of the present application may be HLG-1002-1073, HLG-1002-1166 or HLG-1002-1167; the coptis paste negative lacking coptis sample (batch number HLG-1002-1075), coptis paste negative lacking phellodendron sample (batch number HLG-1002-1076), coptis paste double negative lacking coptis and phellodendron sample (batch number HLG-1002-1078) were prepared by referring to the prescription and method of the coptis paste; coptis control medicinal materials and phellodendron control medicinal materials were purchased from the China Food and Drug Inspection Institute; berberine hydrochloride was purchased from the China Food and Drug Inspection Institute; the silica gel G thin layer plate used in the present application was purchased from Qingdao Ocean Chemical Plant Branch, Yantai Huayang New Materials Technology Co., Ltd., Yantai Jiangyou Silica Gel Development Co., Ltd. or Merck, Germany. The raw materials, reagents, solvents, etc. used in the examples of the present application can all be obtained commercially.

[0022] The present application is further described in detail below in conjunction with embodiments, test experiments and accompanying drawings.

[0023] Example 1 Embodiment 1 provides a thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense, comprising the following steps: (1) Prepare the test sample solution, Rhizoma Coptidis reference medicinal material solution, Rhizoma Phellodendri reference medicinal material solution and reference substance solution respectively; (1-1) Test solution: add 30 mL of water to 10 g of the sample to be tested, heat to melt, then heat under reflux and stir at 420 rpm for 30 min, cool to room temperature; then store at 0±5°C for 30 min, filter, add 60 mL of ethyl acetate to the filtrate for extraction, collect the aqueous phase, evaporate to dryness, add 1 mL of methanol to dissolve it, and obtain the test solution; (1-2) Coptis chinensis control medicinal material solution: The preparation method of Coptis chinensis control medicinal material solution is as follows: add 30 mL of water to 20 mg of Coptis chinensis control medicinal material, then heat and reflux at 420 rpm for 30 min, and cool to room temperature; after filtering, add 60 mL of ethyl acetate to the filtrate for extraction, collect the aqueous phase and evaporate to dryness, add 1 mL of methanol to dissolve it, and obtain Coptis chinensis control medicinal material solution; (1-3) Phellodendron chinense control medicinal material solution: The preparation method of Phellodendron chinense control medicinal material solution is as follows: add 30 mL of water to 20 mg of Phellodendron chinense control medicinal material, then heat and reflux at 420 rpm for 30 min, and cool to room temperature; after filtering, add 60 mL of ethyl acetate to the filtrate for extraction, collect the aqueous phase and evaporate to dryness, add 1 mL of methanol to dissolve it, and obtain Phellodendron chinense control medicinal material solution; (1-4) Take 0.05 mg of berberine hydrochloride reference substance, add 1 mL of methanol to dissolve it, and obtain a reference substance solution with a concentration of 0.05 mg / mL; (2) Take 5 μL of each of the above four solutions and spot them on the same silica gel G thin layer plate (Qingdao Ocean, 25°C, 67% humidity). Use n-hexane-n-butanol-methanol-water with a volume ratio of 3:3:2:1 as the developing solvent. Place the silica gel G thin layer plate in a developing cylinder that has been saturated with concentrated ammonia solution for 20-30 minutes, develop, take out, dry, and then examine under a 365nm light source.

[0024] Exclusive inspection The samples of Coptis chinensis paste, Coptis chinensis paste negative lacking Coptis chinensis, Coptis chinensis paste negative lacking Phellodendron amurense, and Coptis chinensis paste double negative lacking Coptis chinensis and Phellodendron amurense were used as test samples, and the test was carried out according to the method of Example 1. The results are as follows: Figure 1 shown.

[0025] Figure 1 This is a thin layer chromatogram showing the specificity investigation results. Figure 1 Among them, 1 is berberine hydrochloride reference solution; 2 is coptis chinensis reference medicinal material solution; 3 is phellodendron chinense reference medicinal material solution; 4 is HLG-1002-1073 coptis chinensis paste test solution; 5 is HLG-1002-1075 coptis chinensis paste negative test solution lacking coptis chinensis; 6 is HLG-1002-1076 coptis chinensis paste negative test solution lacking phellodendron chinense; 7 is HLG-1002-1078 coptis chinensis paste double negative test solution lacking coptis chinensis and phellodendron chinense).

[0026] Depend on Figure 1 It can be seen that the separation between the components in the test sample is good, and there is no interference from other impurities in the chromatogram of the test sample solution of the double negative of the coptis paste lacking coptis and phellodendron, which indicates that the thin layer chromatography method provided by the present application for simultaneously identifying the components of coptis and phellodendron has good specificity.

[0027] Durability test (1) Durability inspection of thin layer board manufacturers The separation effect of silica gel G thin layer plates produced by three manufacturers, Merck, Yantai Huayang and Yantai Jiangyou, on coptis paste was investigated. According to the method in Example 1, berberine hydrochloride reference solution, coptis chinensis reference medicinal material solution, phellodendron chinense reference medicinal material solution, coptis chinensis paste test solution 1 (batch number HLG-1002-1166) and coptis chinensis paste test solution 2 (batch number HLG-1002-1167) were prepared respectively, and the above solutions were spotted on the above three silica gel G thin layer plates, and separation and detection were carried out according to the method in Example 1. The results are as follows: Figure 2 shown.

[0028] Figure 2 It is a thin layer chromatogram obtained by using silica gel G thin layer plates produced by different manufacturers; Figure 2 In the figure, (a) is the German Merck manufacturer, (b) is the Yantai Huayang manufacturer, and (c) is the Yantai Jiangyou manufacturer; in each chromatogram, from left to right: 1 is berberine hydrochloride reference solution; 2 is coptis chinensis reference medicinal material solution; 3 is phellodendron chinense reference medicinal material solution; 4 is HLG-1002-1166 coptis chinensis paste test solution 1; 5 is HLG-1002-1167 coptis chinensis paste test solution 2.

[0029] Figure 2 The results show that the silica gel G thin layer plates produced by Merck, Yantai Huayang and Yantai Jiangyou can effectively separate the components in the coptis paste with good separation. Therefore, it is shown that the thin layer chromatography method for simultaneously identifying the components of coptis and phellodendron provided by this application has good durability.

[0030] (2) Environmental durability assessment The separation effects of silica gel G thin layer plates produced by Qingdao Ocean and Yantai Jiangyou on coptis paste were investigated under low temperature of 4°C and humidity of 92%. According to the method in Example 1, berberine hydrochloride reference solution, coptis control medicinal material solution, phellodendron control medicinal material solution, HLG-1002-1073 coptis paste test solution, HLG-1002-1075 coptis paste negative test solution lacking coptis, HLG-1002-1076 coptis paste negative test solution lacking phellodendron, and HLG-1002-1078 coptis paste double negative test solution lacking coptis and phellodendron were prepared respectively. The above solutions were spotted on the above four silica gel G thin layer plates, and separation and detection were carried out according to the method in Example 1. The results are as follows. Figure 3 shown.

[0031] Figure 3 Thin layer chromatograms obtained for silica gel G thin layer plate detection under different environmental conditions; Figure 3Among them, (a) is Qingdao Ocean manufacturer, 4℃ low temperature, 67% humidity, (b) is Qingdao Ocean manufacturer, 25℃, 92% humidity, (c) is Yantai Jiangyou manufacturer, 4℃ low temperature, 67% humidity, (d) is Yantai Jiangyou manufacturer, 25℃, 92% humidity; from left to right in each chromatogram: 1 is berberine hydrochloride reference solution; 2 is Coptis chinensis reference medicinal material solution; 3 is Phellodendron chinense reference medicinal material solution; 4 is HLG-1002-1073 Coptis chinensis paste test solution; 5 is HLG-1002-1075 Coptis chinensis paste negative lack of Coptis chinensis test solution; 6 is HLG-1002-1076 Coptis chinensis paste negative lack of Phellodendron chinense test solution; 7 is HLG-1002-1078 Coptis chinensis paste double negative lack of Coptis chinensis and Phellodendron chinense test solution.

[0032] Figure 3 The results show that the silica gel G thin layer plates produced by Qingdao Ocean and Yantai Jiangyou can effectively separate the components in the coptis paste at a low temperature of 4°C and a humidity of 92%, and the separation degree is good. Therefore, it is shown that the thin layer chromatography method for simultaneously identifying the components of coptis and phellodendron provided by this application has good durability.

[0033] Embodiment 2-7 Examples 2-7 respectively provide a thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense.

[0034] The difference between the above embodiment and embodiment 1 is that the ratio of the developing agent is as shown in Table 1 below.

[0035] Comparative Examples 1-5 Comparative Examples 1-5 respectively provide a thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense.

[0036] The difference between the comparative example and Example 1 is: the type and ratio of the developing agent, as shown in Table 1 below.

[0037] Table 1 Types and ratios of developing agents used in Examples 2-7 and Comparative Examples 1-5 Test results The HLG-1002-1073 Coptis chinensis paste test solution was tested according to the thin layer chromatography method provided in Examples 1-7 and Comparative Examples 1-5, and the separation effect, the number of spots and the ratio shift value RF value in the chromatogram were obtained. The results are shown in Table 2 below.

[0038] (1) Separation effect: Observe whether the spots of the test solution of Coptis chinensis paste are clear and bright, and whether the shapes are regular. Count the number of characteristic spots and check whether there are impurity spots.

[0039] (2) Ratio shift value (RF value): the ratio of the distance from the origin to the center of the main spot to the distance from the origin to the solvent front; Note: The RF value is in the reasonable range of 0.2-0.8.

[0040] Table 2 Detection results of Huanglian ointment obtained by thin layer chromatography method of Examples 1-7 and Comparative Examples 1-5 According to the test results in Table 2, in the thin layer chromatography method provided in Examples 1-7 of the present application, n-hexane-n-butanol-methanol-water with a volume ratio of (2.5-3.5): (2.5-3.5): (1.5-2.5): (0.8-1.2) is used as the developing solvent, and the obtained chromatogram has 4 obvious characteristic spots, and the spots are clear, bright, regular, and free of impurity spots. Therefore, it is shown that the thin layer chromatography method provided in the present application can realize the simultaneous identification of the components of Coptis chinensis and Phellodendron chinense in Coptis chinensis paste, and the separation and identification effect is good.

[0041] In the thin layer chromatography method provided in Comparative Example 1, n-hexane-n-butanol-methanol-water with a volume ratio of 4:4:3:1 was used as the developing solvent, and the number of characteristic spots in the obtained chromatogram was only 3, and there were impurity spots. Therefore, it is shown that the above method cannot simultaneously identify the components of Coptis chinensis and Phellodendron chinense in Coptis chinensis paste.

[0042] In the thin layer chromatography method provided in Comparative Example 2, n-butanol-methanol-water with a volume ratio of 3:2:1 was used as the developing solvent, and the number of characteristic spots in the obtained chromatogram was only 3, and there were impurity spots. Therefore, it was shown that the above method could not simultaneously identify the components of Coptis chinensis and Phellodendron chinense in Coptis chinensis paste.

[0043] In the thin layer chromatography method provided in Comparative Example 3, n-hexane-methanol-water with a volume ratio of 3:2:1 was used as the developing solvent, and the number of characteristic spots in the obtained chromatogram was only 2, and there were impurity spots. Therefore, it was shown that the above method could not simultaneously identify the components of Coptis chinensis and Phellodendron chinense in Coptis chinensis paste.

[0044] In the thin layer chromatography method provided in Comparative Example 4, chloroform-methanol-water with a volume ratio of 10:5:1 was used as the developing solvent, and the number of characteristic spots in the obtained chromatogram was only 1, and there were impurity spots. Therefore, it was shown that the above method could not simultaneously identify the components of Coptis chinensis and Phellodendron chinense in Coptis chinensis paste.

[0045] In the thin layer chromatography method provided in Comparative Example 5, n-butanol-acetic acid-water with a volume ratio of 3:2:1 and a volume ratio of 7:1:2 were used as the developing solvent, and the number of characteristic spots in the obtained chromatogram was only 1, and there were impurity spots. Therefore, it is shown that the above method cannot simultaneously identify the components of Coptis chinensis and Phellodendron chinense in Coptis chinensis paste.

[0046] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A thin layer chromatography method for simultaneously identifying the components of Coptis chinensis and Phellodendron chinense, characterized in that: The following steps are involved: (1) Prepare the test sample solution, Rhizoma Coptidis reference medicinal material solution, Rhizoma Phellodendri reference medicinal material solution and reference substance solution respectively; (2) Spot the above solutions on the same silica gel G thin layer plate respectively, use n-hexane-n-butanol-methanol-water with a volume ratio of (2.5-3.5):(2.5-3.5):(1.5-2.5):(0.8-1.2) as the developing agent, place the silica gel G thin layer plate in a developing tank for development, and examine it under a light source of 360-370nm.

2. The thin layer chromatography method according to claim 1, characterized in that The volume ratio of n-hexane-n-butanol-methanol-water is 3:3:2:

1.

3. The thin layer chromatography method according to claim 1, characterized in that: The spotting volume of each solution was 3-8 μL.

4. The thin layer chromatography method according to claim 1, characterized in that The developing cylinder is pre-saturated with concentrated ammonia solution for 20-30 minutes.

5. The thin layer chromatography method according to claim 1, characterized in that: The preparation method of the test solution is as follows: add water to the sample to be tested, heat to melt, then heat under reflux and stir at 400-450 rpm for 30-40 min, cool to room temperature; then store at 0±5°C for 25-35 min, filter, extract, evaporate the water phase, add methanol to dissolve, and obtain the test solution.

6. The thin layer chromatography method according to claim 1, characterized in that: The sample to be tested is Coptis chinensis paste.

7. The thin layer chromatography method according to claim 1, characterized in that: The preparation method of the Coptidis reference medicinal material solution is as follows: water is added to the Coptidis reference medicinal material, and then heated under reflux and stirred at 400-450 rpm for 30-40 minutes, and cooled to room temperature; filtering, extracting, evaporating the water phase, and adding methanol to dissolve to obtain the Coptidis reference medicinal material solution.

8. The thin layer chromatography method according to claim 1, characterized in that: The preparation method of the Phellodendron chinense reference medicinal material solution is as follows: water is added to the Phellodendron chinense reference medicinal material, and then heated under reflux and stirred at 400-450 rpm for 30-40 minutes, and cooled to room temperature; filtering, extracting, evaporating the water phase, and adding methanol to dissolve to obtain the Phellodendron chinense reference medicinal material solution.

9. The thin layer chromatography method according to any one of claims 1 to 8, characterized in that The preparation method of the reference solution is as follows: take berberine hydrochloride reference, add methanol to dissolve it, and obtain a reference solution with a concentration of 0.05 mg / mL.

10. Use of the thin layer chromatography method for simultaneously identifying the components of coptis chinensis and phellodendron chinense as claimed in any one of claims 1 to 9 in identifying the components of coptis chinensis and phellodendron chinense in traditional Chinese medicine external preparations.

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