Thin-layer chromatography method for simultaneously identifying multi-index components in cassia twig and ginseng decoction
By optimizing the preparation methods of test samples and control medicinal materials, and combining specific thin-layer plate and developing solvent color development techniques, we have achieved efficient and accurate identification of multiple components in Guizhi Renshen Decoction, solving the problem of difficulty in simultaneously identifying multiple indicator components in existing technologies, and reducing equipment costs and environmental control requirements.
Patent Information
- Application Number
- CN202511286121.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-10
- Publication Date
- 2025-10-31
AI Technical Summary
Existing technologies make it difficult to quickly and accurately identify multiple active ingredients in Guizhi Renshen Decoction simultaneously, and traditional methods require complex sample pretreatment and expensive instruments and equipment.
By optimizing the preparation methods of the test sample and reference medicinal materials, and combining silica gel G thin-layer plate, chloroform-petroleum ether-ethyl acetate developing solvent and 365nm ultraviolet light color development, a thin-layer chromatography method was established to achieve simultaneous identification of multiple index components in Guizhi Renshen Decoction.
It improves the separation and detection sensitivity of each component in Guizhi Renshen Decoction, with clear color development and no interference. Moreover, the thin-layer chromatogram is stable within a certain temperature and humidity range, reducing the need for environmental control and improving the accuracy of identification results.
Smart Images
Figure CN120870433A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of traditional Chinese medicine quality control technology, and in particular to a thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction. Background Technology
[0002] Guizhi Renshen Tang (Cinnamon Twig and Ginseng Decoction) is a classic formula from the *Shanghan Lun* (Treatise on Cold Damage), composed of cinnamon twig, ginseng, atractylodes macrocephala, dried ginger, and prepared licorice root. It has the effects of warming the middle jiao (spleen and stomach), relieving exterior syndromes, strengthening the spleen, and replenishing qi. Clinically, it is often used to treat exogenous wind-cold syndrome complicated by spleen and stomach deficiency-cold. Modern research shows that the pharmacological effects of this formula are closely related to its various active ingredients, such as cinnamaldehyde, ginsenosides, and glycyrrhizic acid in cinnamon twig. Therefore, establishing a rapid, accurate, and reliable method to simultaneously identify multiple indicator components in Guizhi Renshen Tang is of great significance for ensuring its quality control, clinical efficacy, and further research.
[0003] Currently, commonly used methods for component analysis in traditional Chinese medicine (TCM) compound prescriptions include high-performance liquid chromatography (HPLC), gas chromatography (GC), and liquid chromatography-mass spectrometry (LC-MS). However, these methods typically require complex sample pretreatment, expensive equipment, and lengthy analysis times. In contrast, thin-layer chromatography (TLC) offers advantages such as ease of operation, low cost, and the ability to analyze multiple samples simultaneously, making it particularly suitable for the preliminary identification and quality control of TCM compound prescriptions. However, existing research largely focuses on TLC detection of single components, and a systematic report on TLC methods for the simultaneous identification of multiple components in Guizhi Renshen Tang (Cinnamon Twig and Ginseng Decoction) has not yet been found.
[0004] This invention aims to establish an optimized thin-layer chromatography method that can simultaneously identify cinnamon twig, ginseng, atractylodes macrocephala, dried ginger, and roasted licorice root in Guizhi Renshen Decoction. Summary of the Invention
[0005] To address the above shortcomings, this invention provides a thin-layer chromatography method for simultaneously identifying multiple components in Guizhi Renshen Decoction. By optimizing the preparation methods of the test sample and reference herbs, and optimizing the developing solvent system and color development conditions, the method improves the separation and detection sensitivity of each component in Guizhi Renshen Decoction, resulting in good separation, clear color development, and no interference. Furthermore, the thin-layer chromatograms exhibit good separation performance under different temperature and humidity conditions, eliminating the need for strict environmental control. The specific technical solution is as follows: A thin-layer chromatography method for simultaneously identifying multiple components in Guizhi Renshen Decoction includes the following steps: (1) Preparation of test solution: Take the sample of Guizhi Renshen Decoction to be tested, extract with ethyl acetate, evaporate the extract to dryness, dissolve the residue with methanol to obtain the test solution; (2) Preparation of reference medicinal material solution: Take ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials respectively, decoct them with water, filter them, concentrate the filtrate, extract with ethyl acetate, evaporate the extract to dryness, add methanol to dissolve the residue to obtain reference medicinal material solution; (3) Spotting, development and inspection: Take the test sample solution and the reference medicinal material solution respectively and spot them on the same thin layer plate. Develop with chloroform-petroleum ether-ethyl acetate as the developing solvent, dry, and first inspect under a 365nm ultraviolet lamp. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Atractylodes macrocephala reference medicinal material and Cinnamomum cassia reference medicinal material. Then spray with 5% vanillin sulfuric acid solution and heat at 105℃ until the spots are clearly visible. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Ginseng reference medicinal material, Glycyrrhiza uralensis reference medicinal material and Zingiber officinale reference medicinal material.
[0006] Preferably, in step (1), the preparation method of the sample of Guizhi Renshen Decoction to be tested is as follows: weigh 40-45g each of ginseng, atractylodes macrocephala and dried ginger, and 50-60g each of cinnamon twig and roasted licorice root. First, put the ginseng, atractylodes macrocephala, dried ginger and roasted licorice root into a pot, add 1500-1800mL of water, bring to a boil over high heat and then simmer over low heat until 800-1000mL of water is obtained. Add cinnamon twig and continue to simmer until 500-600mL of water is obtained. Remove the pot and filter while hot to obtain the sample of Guizhi Renshen Decoction.
[0007] Preferably, in step (1), the preparation of the test solution is as follows: take 15-25 mL of the Guizhi Renshen Decoction sample to be tested, add ethyl acetate and shake to extract 2-3 times, 25-35 mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1 mL of methanol to dissolve the residue, and obtain the test solution.
[0008] Preferably, in step (2), the preparation of the reference medicinal material solution is as follows: take 1-5g of each of ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials, add 30-100mL of water and decoct for 1-2 hours, filter, concentrate the filtrate to 20-80mL, add ethyl acetate and shake to extract 2-3 times, 25-35mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1mL of methanol to dissolve the residue to obtain the reference medicinal material solution.
[0009] Preferably, in step (3), 1-3 μL of the test solution and the control medicinal material solution are spotted on the same thin-layer plate.
[0010] Preferably, in step (3), the thin film is a silicone G thin film.
[0011] Preferably, in step (3), the ratio of chloroform:petroleum ether:ethyl acetate in the developing agent is 10:5:1.
[0012] Preferably, in step (3), the temperature during the unfolding of the developing agent is 10℃-40℃ and the relative humidity is 25%-75%.
[0013] Compared with the prior art, the beneficial effects of the present invention are: 1. This invention optimizes the preparation methods of the test sample and reference medicinal materials, and, together with silica gel G thin-layer plate, developing solvent (chloroform-petroleum ether-ethyl acetate), 365nm ultraviolet lamp, and colorimetric agent 5% vanillin sulfuric acid solution, constitutes a new thin-layer identification method. This improves the separation and detection sensitivity of each component in Guizhi Renshen Decoction, resulting in good separation of each component, clear color development, and no interference. It enables the simultaneous identification of components such as ginseng, roasted licorice, dried ginger, cinnamon twig, and atractylodes macrocephala in Guizhi Renshen Decoction.
[0014] 2. The thin-layer chromatograms of this invention are less affected by environmental interference and exhibit good separation effects under conditions of 10~40℃ and 25%~75% humidity, without the need for strict environmental control, thus improving the accuracy of the identification results. Attached Figure Description
[0015] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below.
[0016] Figure 1 The thin-layer chromatogram of Example 3; Figure 2 This is the thin-layer chromatogram of Comparative Example 1; Figure 3 This is the thin-layer chromatogram of Comparative Example 2; Figure 4 This is the thin-layer chromatogram of Comparative Example 3; Figure 5 This is the thin-layer chromatogram of Comparative Example 4; Figure 6 This is the thin-layer chromatogram of Comparative Example 5; Figure 7 This is the thin-layer chromatogram of Comparative Example 6; Figure 8 The image shows a thin-layer chromatogram prepared using silica gel G plates from Qingdao Marine Chemical Co., Ltd. Figure 9 The thin-layer chromatogram was prepared using Yantai Xinnuo silica gel G plates. Figure 10 The image shows a thin-layer chromatogram prepared using a high-efficiency silica gel plate from Qingdao. Figure 11 The image shows a thin-layer chromatogram prepared at 10℃ and 75% relative humidity. Figure 12 The image shows a thin-layer chromatogram prepared at 25℃ and 75% relative humidity. Figure 13The thin-layer chromatogram was prepared at 40℃ and 75% relative humidity. Figure 14 The image shows a thin-layer chromatogram prepared at 25℃ and 25% relative humidity. Figure 15 The image shows a thin-layer chromatogram prepared at 25°C and 50% relative humidity. Detailed Implementation
[0017] The specific embodiments of the present invention will be described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments.
[0018] Example 1 This embodiment presents a thin-layer chromatography method for simultaneously identifying multiple components in Guizhi Renshen Decoction, comprising the following steps: (1) Preparation of test solution: Take 15 mL of the Guizhi Renshen Decoction sample to be tested, add ethyl acetate and shake to extract twice, 25 mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1 mL of methanol to dissolve the residue to obtain the test solution.
[0019] The preparation method of the Guizhi Renshen Decoction sample to be tested is as follows: Weigh 40g each of ginseng, atractylodes macrocephala and dried ginger, and 50g each of cinnamon twig and prepared licorice root. First, put the ginseng, atractylodes macrocephala, dried ginger and prepared licorice root into a pot, add 1500mL of water, bring to a boil over high heat, then simmer over low heat until 800mL of water remains. Add cinnamon twig and continue to simmer until 500mL of water remains. Remove from heat, filter while hot, and the Guizhi Renshen Decoction sample is obtained.
[0020] (2) Preparation of the reference medicinal material solution: Take 1g of each of ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials, add 30mL of water and decoct for 1 hour, filter, concentrate the filtrate to 20mL, add ethyl acetate and shake to extract twice, 25mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1mL of methanol to dissolve the residue to obtain the reference medicinal material solution.
[0021] (3) Spotting, development and inspection: Take 1 μL of the test sample solution and the reference medicinal material solution respectively and spot them on the same silica gel G thin layer plate. Develop with chloroform-petroleum ether-ethyl acetate (10:5:1) as the developing solvent. The development temperature is 10℃ and the relative humidity is 25%. After drying, inspect under a 365nm ultraviolet lamp. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Atractylodes macrocephala reference medicinal material and Cinnamomum cassia reference medicinal material. Then spray with 5% vanillin sulfuric acid solution and heat at 105℃ until the spots are clearly visible. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Ginseng reference medicinal material, Glycyrrhiza uralensis reference medicinal material and Zingiber officinale reference medicinal material.
[0022] This embodiment can simultaneously identify the five components of Guizhi Renshen Decoction: ginseng, roasted licorice root, dried ginger, cinnamon twig, and atractylodes macrocephala. The separation of each component is good, the color development is clear, and there is no interference.
[0023] Example 2 This embodiment presents a thin-layer chromatography method for simultaneously identifying multiple components in Guizhi Renshen Decoction, comprising the following steps: (1) Preparation of test solution: Take 25 mL of the sample of Guizhi Renshen Decoction to be tested, add ethyl acetate and shake to extract 3 times, 35 mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1 mL of methanol to dissolve the residue to obtain the test solution.
[0024] The preparation method of the Guizhi Renshen Decoction sample to be tested is as follows: Weigh 45g each of ginseng, atractylodes macrocephala and dried ginger, and 60g each of cinnamon twig and prepared licorice root. First, put the ginseng, atractylodes macrocephala, dried ginger and prepared licorice root into a pot, add 1800mL of water, bring to a boil over high heat, then simmer over low heat until 1000mL of water remains. Add cinnamon twig and continue to simmer until 600mL of water remains. Remove from heat, filter while hot, and the Guizhi Renshen Decoction sample is obtained.
[0025] (2) Preparation of the reference medicinal material solution: Take 5g of each of ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials, add 100mL of water and decoct for 2 hours, filter, concentrate the filtrate to 80mL, add ethyl acetate and shake to extract 3 times, 35mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1mL of methanol to dissolve the residue to obtain the reference medicinal material solution.
[0026] (3) Spotting, development and inspection: Take 3 μL of the test solution and the reference medicinal material solution respectively and spot them on the same silica gel G thin layer plate. Develop with chloroform-petroleum ether-ethyl acetate (10:5:1) as the developing solvent. The development temperature is 40℃ and the relative humidity is 75%. After drying, inspect under a 365nm ultraviolet lamp. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Atractylodes macrocephala and Cinnamomum cassia reference medicinal materials. Then spray with 5% vanillin sulfuric acid solution and heat at 105℃ until the spots are clearly visible. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Ginseng, Glycyrrhiza uralensis, and Zingiber officinale reference medicinal materials.
[0027] This embodiment can simultaneously identify the five components of Guizhi Renshen Decoction: ginseng, roasted licorice root, dried ginger, cinnamon twig, and atractylodes macrocephala. The separation of each component is good, the color development is clear, and there is no interference.
[0028] Example 3 This embodiment presents a thin-layer chromatography method for simultaneously identifying multiple components in Guizhi Renshen Decoction, comprising the following steps: (1) Preparation of test solution: Take 20 mL of the sample of Guizhi Renshen Decoction to be tested, add ethyl acetate and shake to extract twice, 30 mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1 mL of methanol to dissolve the residue to obtain the test solution.
[0029] The preparation method of the Guizhi Renshen Decoction sample to be tested is as follows: Weigh 41g each of ginseng, atractylodes macrocephala and dried ginger, and 55g each of cinnamon twig and prepared licorice root. First, put the ginseng, atractylodes macrocephala, dried ginger and prepared licorice root into a pot, add 1800mL of water, bring to a boil over high heat, then simmer over low heat until 1000mL of water remains. Add cinnamon twig and continue to simmer until 600mL of water remains. Remove from heat, filter while hot, and the Guizhi Renshen Decoction sample is obtained.
[0030] (2) The preparation of the reference medicinal material solution is as follows: Take 1g of each of ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials, add 30mL of water and decoct for 1 hour, filter, concentrate the filtrate to 20mL, add ethyl acetate and shake to extract twice, 30mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1mL of methanol to dissolve the residue to obtain the reference medicinal material solution.
[0031] (3) Spotting, development, and inspection: Take 3 μL each of the test solution and the reference herb solution and spot them on the same silica gel G thin-layer plate. Develop with chloroform-petroleum ether-ethyl acetate (10:5:1) as the developing solvent at 25℃ and 64% relative humidity. After drying, inspect under a 365nm UV lamp. The test sample should show spots of the same color at the corresponding positions as the Atractylodes macrocephala and Cinnamomum cassia reference herbs (see thin-layer chromatogram). Figure 1 (As shown in A), then spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. The test sample shows spots of the same color at the corresponding positions as those of ginseng reference material, licorice reference material, and dried ginger reference material (thin-layer chromatogram as shown in A). Figure 1 As shown in B).
[0032] This embodiment can simultaneously identify the five components of Guizhi Renshen Decoction: ginseng, roasted licorice root, dried ginger, cinnamon twig, and atractylodes macrocephala. The separation of each component is good, the color development is clear, and there is no interference.
[0033] Comparative Example 1: The steps of this comparative example are as follows: (1) Preparation of the test solution: Take 20 mL of the Guizhi Renshen Decoction sample to be tested, add 20 mL of water-saturated n-butanol, and extract twice by shaking, 20 mL each time. Combine the n-butanol extracts and extract twice by shaking with ammonia solution, 20 mL each time. Evaporate the n-butanol solution to dryness, and dissolve the residue in 2 mL of methanol to obtain the test solution. The preparation method of the Guizhi Renshen Decoction sample to be tested is the same as in Example 3.
[0034] (2) The preparation of the reference medicinal material solution is as follows: take 1g of each of ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials, add 3mL of methanol, soak overnight in cold water, and take the supernatant to obtain the reference medicinal material solution.
[0035] (3) Spotting, development, and inspection: Take 3 μL each of the test solution and the reference herb solution and spot them onto the same silica gel G thin-layer plate. Develop with toluene:acetone (2:1) as the developing solvent at 25℃ and 50% relative humidity. After drying, inspect under a 365nm UV lamp. The thin-layer chromatogram is shown below. Figure 2 As shown.
[0036] Comparative Example 2: Except for spraying with 5% vanillin-sulfuric acid solution and heating at 105℃ to develop the spots, the other steps in this comparative example were the same as in Comparative Example 1. Its thin-layer chromatogram is shown below. Figure 3 As shown.
[0037] According to Example 3 ( Figure 1 Comparative Example 1 Figure 2 ) and Comparative Example 2 ( Figure 3 Thin-layer chromatograms show that the test solution and the reference herbal solution were prepared using ethyl acetate extraction. The resulting chromatograms exhibited a large number of characteristic spots and good separation, providing a more comprehensive reflection of the characteristic information of each component. Simultaneously, through… Figure 1 and Figure 2 , Figure 3 The comparison also shows that the present invention optimizes the preparation methods of the test sample and the reference medicinal materials, and together with silica gel G thin-layer plate, developing solvent (chloroform-petroleum ether-ethyl acetate), 365nm ultraviolet lamp, and color developer 5% vanillin sulfuric acid solution, it constitutes a new thin-layer identification method, which improves the separation degree and detection sensitivity of each component in Guizhi Renshen Decoction, making the separation degree of each component good, the color development clear, and the absence of interference, and realizing the simultaneous identification of ginseng, roasted licorice, dried ginger, cinnamon twig, atractylodes macrocephala and other components in Guizhi Renshen Decoction.
[0038] Comparative Example 3: The developing solvent for this comparative example was petroleum ether:ethyl acetate:formic acid (6:4:0.1). Other steps were the same as in Example 3. The thin-layer chromatogram is shown below. Figure 4 As shown, Figure 4 A represents the results of the examination under ultraviolet light. Figure 4 B represents the examination results under the chromogenic reagent of 5% vanillin sulfuric acid solution.
[0039] Comparative Example 4: The developing solvent for this comparative example was chloroform, and the other steps were the same as in Example 3. Its thin-layer chromatogram is shown below. Figure 5 As shown, Figure 5 A represents the results of the examination under ultraviolet light. Figure 5 B represents the examination results under the chromogenic reagent of 5% vanillin sulfuric acid solution.
[0040] Comparative Example 5: The developing solvent for this comparative example was chloroform:petroleum ether:ethyl acetate (10:5:0.5). Other steps were the same as in Example 3. The thin-layer chromatogram is shown below. Figure 6 As shown, Figure 6A represents the results of the examination under ultraviolet light. Figure 6 B represents the examination results under the chromogenic reagent of 5% vanillin sulfuric acid solution.
[0041] Comparative Example 6: The developing solvent for this comparative example was chloroform:petroleum ether:ethyl acetate (10:0.3:0.2). Other steps were the same as in Example 3. The thin-layer chromatogram is shown below. Figure 7 As shown, Figure 7 A represents the results of the examination under ultraviolet light. Figure 7 B represents the examination results under the chromogenic reagent of 5% vanillin sulfuric acid solution.
[0042] Based on the thin-layer chromatograms of Examples 3 and Comparative Examples 3 to 6, the present invention uses chloroform:petroleum ether:ethyl acetate (10:5:1) as the developing ratio for thin-layer chromatographic identification of Guizhi Renshen Decoction. The test sample showed clear, bright, and regular spots in the five reference herbs: ginseng, roasted licorice, dried ginger, cinnamon twig, and atractylodes macrocephala, with no impurity spots.
[0043] Durability assessment of thin-layer board manufacturers Investigating high-efficiency silicone sheets from different manufacturers: Qingdao Haiyang Sheet (Qingdao Haiyang Chemical Silicone G Sheet) Figure 8 Yantai Jiangyou Board (Yantai Xinnuo Silicone G Board) Figure 9 Qingdao High-Efficiency Silicone Sheet ( Figure 10 The effect of thin-layer chromatography on Guizhi Renshen Decoction was investigated. Other identification methods were the same as in Example 3, and the results are as follows: Figures 8 to 10 As shown in the figure, silica gel G plates from different manufacturers all exhibit good separation effects for the thin-layer chromatography of Guizhi Renshen Tang, indicating that the identification of this invention does not require special selection of the thin-layer plate.
[0044] Environmental durability study of thin-layer plates The effects of different developing temperatures (10℃, 25℃, 40℃) and different relative humidity (25%, 50%, 75%) on the thin-layer chromatography behavior were investigated. Other identification methods were the same as in Example 3. The results are as follows: Figures 11 to 15 As shown, Figure 11 The spectrum is at 10℃ and 75% relative humidity. Figure 12 The spectrum is at 25℃ and 75% relative humidity. Figure 13 The spectrum is at 40℃ and 75% relative humidity. Figure 14 The spectrum is at 25℃ and 25% relative humidity. Figure 15 The chromatograms are at 25℃ and 50% relative humidity. The figures show that the thin-layer chromatograms of Guizhi Renshen Decoction under different temperature and humidity conditions all have good separation effects, indicating that thin-layer chromatographic identification can be performed at temperatures of 10~40℃ and humidity of 25%~75%, without the need for strict control.
[0045] In summary, this invention optimizes the preparation methods of the test sample and reference medicinal materials, and, together with silica gel G thin-layer plate, developing solvent (chloroform-petroleum ether-ethyl acetate), 365nm ultraviolet lamp, and colorimetric reagent 5% vanillin sulfuric acid solution, constitutes a new thin-layer identification method. This improves the separation and detection sensitivity of each component in Guizhi Renshen Decoction, resulting in good separation, clear color development, and no interference among the components. It enables the simultaneous identification of components such as ginseng, roasted licorice, dried ginger, cinnamon twig, and atractylodes macrocephala in Guizhi Renshen Decoction.
[0046] Meanwhile, the thin-layer chromatograms of this invention are less affected by environmental interference and have good separation effects under conditions of temperature 10℃~40℃ and humidity 25%~75%, without the need for strict environmental control, thus improving the accuracy of the identification results.
[0047] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.
Claims
1. A thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction, characterized in that, Includes the following steps: (1) Preparation of test solution: Take the sample of Guizhi Renshen Decoction to be tested, extract with ethyl acetate, evaporate the extract to dryness, dissolve the residue with methanol to obtain the test solution; (2) Preparation of reference medicinal material solution: Take ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials respectively, decoct them with water, filter them, concentrate the filtrate, extract with ethyl acetate, evaporate the extract to dryness, add methanol to dissolve the residue to obtain reference medicinal material solution; (3) Spotting, development and inspection: Take the test solution and the reference medicinal material solution respectively and spot them on the same thin layer plate. Develop with chloroform-petroleum ether-ethyl acetate as the developing solvent, dry, and first inspect under a 365nm ultraviolet lamp. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Atractylodes macrocephala reference medicinal material and Cinnamomum cassia reference medicinal material. Then spray with 5% vanillin sulfuric acid solution and heat until the spots are clearly visible. The test sample shows spots of the same color at the corresponding positions of the chromatograms of Ginseng reference medicinal material, Glycyrrhiza uralensis reference medicinal material and Zingiber officinale reference medicinal material.
2. The thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction according to claim 1, characterized in that, In step (1), the preparation method of the sample of Guizhi Renshen Decoction to be tested is as follows: weigh 40-45g each of ginseng, atractylodes macrocephala and dried ginger, and 50-60g each of cinnamon twig and prepared licorice root. First, put the ginseng, atractylodes macrocephala, dried ginger and prepared licorice root into a pot, add 1500-1800mL of water, bring to a boil over high heat and then simmer over low heat until 800-1000mL of water remains. Add cinnamon twig and continue to simmer until 500-600mL of water remains. Remove the pot, filter while hot, and the Guizhi Renshen Decoction sample is obtained.
3. The thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction according to claim 1, characterized in that, In step (1), the preparation of the test solution is as follows: take 15-25 mL of the Guizhi Renshen Decoction sample to be tested, add ethyl acetate and shake to extract 2-3 times, 25-35 mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1 mL of methanol to dissolve the residue, and obtain the test solution.
4. The thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction according to claim 1, characterized in that, In step (2), the preparation of the reference medicinal material solution is as follows: take 1-5g of each of ginseng, licorice, dried ginger, cinnamon twig and atractylodes macrocephala reference medicinal materials, add 30-100mL of water and decoct for 1-2 hours, filter, concentrate the filtrate to 20-80mL, add ethyl acetate and shake to extract 2-3 times, 25-35mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1mL of methanol to dissolve the residue to obtain the reference medicinal material solution.
5. The thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction according to claim 1, characterized in that, In step (3), 1-3 μL of each of the test solution and the reference medicinal material solution are spotted on the same thin-layer plate.
6. The thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction according to claim 1, characterized in that, In step (3), the thin film is a silicone G thin film.
7. The thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction according to claim 1, characterized in that, In step (3), the ratio of chloroform:petroleum ether:ethyl acetate in the developing solvent is 10:5:
1.
8. The thin-layer chromatography method for simultaneously identifying multiple index components in Guizhi Renshen Decoction according to claim 1, characterized in that, In step (3), the temperature during the development of the developing agent is 10℃-40℃ and the relative humidity is 25%-75%.