Thin layer identification method of mugwort and vinegar mugwort formula granules

Abstract

The present invention provides a thin layer identification method for mugwort and vinegar mugwort formula granules, including: taking mugwort formula granules for pretreatment to obtain a test solution; taking vinegar mugwort formula granules for pretreatment to obtain a vinegar mugwort formula granule solution; taking dongguan puer lactone reference substance, adding methanol to dissolve, and obtaining a reference substance solution; the test solution, the vinegar mugwort formula granule solution, and the reference substance solution are subjected to thin layer chromatography, and the thin layer plate is a silica gel G thin layer plate; the developing agent is n-hexane-ethyl acetate-toluene; and the solution is placed under ultraviolet light for inspection. If the test sample and the reference medicinal material solution have fluorescent spots of the same color at the same Rf, and there are fluorescent spots at Rf=0.625-0.741, then it is mugwort formula granules; if there are no fluorescent spots, then it is vinegar mugwort formula granules. By observing the presence of red fluorescent spots at the above-mentioned ratio shift value in the test sample thin layer chromatography as the mugwort formula granule identification point, no red fluorescent spots at the same position are used as the vinegar mugwort formula granule identification point.

Description

Technical Field

[0001] The invention relates to the technical field of analysis and detection, in particular to a thin-layer identification method for mugwort leaves and mugwort vinegar formula granules. Background Art

[0002] Artemisia argyi Lévl. et Vant. is a dried leaf of the Asteraceae plant [1]. It has the effects of warming the meridians and stopping bleeding, dispelling cold and relieving pain, and externally removing dampness and relieving itching. Vinegar-treated wormwood is warm but not dry, focusing on warming the meridians and stopping bleeding, and can enhance the cold-dispelling and analgesic effects. There are differences in the physical and chemical properties and the dehumidifying and relieving itching effects of wormwood before and after processing. The effect of stopping bleeding and warming the interior is better after processing with vinegar. This shows that the chemical composition of wormwood and vinegar-treated wormwood has changed during the processing process.

[0003] The chemical components of mugwort mainly include volatile oils, flavonoids, terpenes, phenylpropanoids, organic acids, steroids, polysaccharides, and trace elements. Current research on mugwort focuses on the composition and pharmacological effects of different processed products. For example, Zhang Tiantian, Sun Lili, Zhou Qian, et al. conducted a GC-MS study on the volatile oil components of mugwort and its processed products, which showed that the chemical composition of mugwort changed significantly after vinegar processing. While properties can be used to distinguish mugwort and vinegar-processed mugwort slices, methods for distinguishing the two formula granules have not been reported.

[0004] Since mugwort and vinegar mugwort granules are made from mugwort through a series of processes, some of which destroy the original properties, the property identification method is not suitable for the identification of formula granules. Currently, there is no effective method for distinguishing mugwort granules from vinegar mugwort granules. Therefore, it is urgent to establish a method that can distinguish mugwort granules from vinegar mugwort granules. Summary of the Invention

[0005] In view of this, the technical problem to be solved by the present invention is to provide a thin layer identification method for mugwort and vinegar mugwort formula granules.

[0006] The terms "include," "comprising," and "having" are used interchangeably herein and are intended to indicate the inclusiveness of a solution, meaning that the solution may contain other elements in addition to the listed elements. It should also be understood that the use of "include," "comprising," and "having" in this document also provides a "consisting of" solution.

[0007] In this application, the term "and / or" describes the association relationship between associated objects, indicating that three relationships may exist. For example, A and / or B can mean: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural.

[0008] In this application, "at least one" means one or more, and "more than one" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items.

[0009] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the following processes does not mean the order of execution. Some or all of the steps can be executed in parallel or sequentially. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0010] The present invention discloses the construction and application of a thin-layer chromatography (TLC) identification method for mugwort and vinegar mugwort granules. By observing the TLC of a sample, the presence of a red fluorescent spot at a ratio shift value of 0.556-0.752 is used as an identification point for the mugwort granules, while the absence of a red fluorescent spot at the same location is used as an identification point for the vinegar mugwort granules. The established TLC method can quickly and effectively identify mugwort and vinegar mugwort granules. This method has strong specificity, good stability, high precision, and is convenient and easy to master. This method enables more standardized quality control of mugwort and vinegar mugwort granules, ensuring the efficacy of the drugs.

[0011] A thin layer identification method for mugwort and mugwort vinegar granules, comprising:

[0012] A) Pre-treat the Artemisia argyi granules to obtain a test solution;

[0013] Pre-treating the vinegar wormwood formula granules to obtain a vinegar wormwood formula granule solution;

[0014] B) Take the scopolamine reference substance and dissolve it in methanol to obtain a reference solution;

[0015] C) The test solution, vinegar wormwood formula granule solution and reference solution were subjected to thin layer chromatography (TLC) using a silica gel G TLC plate and n-hexane-ethyl acetate-toluene as the developing solvent;

[0016] D) Check under ultraviolet light. If the test sample and control medicinal material solution have fluorescent spots of the same color at the same Rf, and there are fluorescent spots at Rf = 0.625-0.741, then they are mugwort formula granules; if there are no fluorescent spots at Rf = 0.625-0.741, then they are vinegar mugwort formula granules.

[0017] The invention provides wormwood leaf and wormwood leaf vinegar formula granules. First, the wormwood leaf formula granules are pretreated to obtain a test solution.

[0018] The pretreatment of the mugwort formula granules or vinegar mugwort formula granules of the present invention is specifically as follows:

[0019] a) adding methanol to the wormwood formula granules or vinegar wormwood formula granules, sonicating, filtering, evaporating, dissolving the residue in water, passing it through a D101 macroporous adsorption resin column, eluting with 30% methanol, discarding the eluate, and then eluting with ethyl acetate, and collecting the eluate;

[0020] b) Recover the solvent from the eluate until it is dry, and dissolve the residue in ethyl acetate to obtain the product.

[0021] The processing method of the mugwort formula granules or the vinegar mugwort formula granules provided by the present invention comprises the following steps: firstly adding methanol to the mugwort formula granules or the vinegar mugwort formula granules, ultrasonicating the granules, filtering, volatilizing, and dissolving the residue in water.

[0022] In some embodiments of the present invention, the ultrasonic time is 5 to 10 minutes.

[0023] In some embodiments of the present invention, the ultrasonic time is 10 minutes.

[0024] Then pass through D101 macroporous adsorption resin column, elute with 30% methanol, discard the eluate, elute with ethyl acetate, and collect the eluate; the inner diameter of the D101 macroporous adsorption resin is 1 cm, and the column height is 14 cm.

[0025] In some embodiments of the present invention, the mass volume ratio of the mugwort formula granules or vinegar mugwort formula granules, methanol, water, 30% methanol and ethyl acetate is 0.5g:10mL:5mL:20mL:20mL.

[0026] The present invention does not limit the specific steps of pre-treating the vinegar and wormwood formula granules, and the same steps may be used as above.

[0027] Take the scopolamine reference substance, add methanol to dissolve it, and obtain a reference substance solution.

[0028] The mass volume ratio of the scopolamine reference substance of the present invention and methanol is 40 μg:1 mL.

[0029] The test solution, vinegar wormwood formula granule solution and reference solution were subjected to thin layer chromatography.

[0030] Preferably, the test solution, vinegar wormwood formula granule solution and reference solution are aspirated and spotted on the same silica gel G thin layer plate, developed with a developing agent, taken out, dried and inspected under ultraviolet light. The ultraviolet light wavelength is 365nm.

[0031] According to the present invention, the developing agent is n-hexane-ethyl acetate-toluene;

[0032] In some preferred embodiments of the present invention, the mass ratio of n-hexane-ethyl acetate-toluene is 1:3:2.

[0033] The thin layer plate described in the present invention is a silica gel G thin layer plate; preferably a GF254 thin layer plate. Prefabricated silica gel GF254 plates can be selected from Tianjin Silida, Merck, and Qingdao Ocean. Results demonstrate that this method is robust and meets identification requirements. Compared with the control herbal material atlas, the sample and control herbal material atlases exhibit spots of the same color at corresponding locations.

[0034] Check under ultraviolet light. If the test and reference solutions have fluorescent spots of the same color at the same Rf, and red fluorescent spots at Rf = 0.625-0.741, then they are mugwort formula granules.

[0035] If there is no fluorescent spot at Rf=0.625~0.741, it is vinegar and mugwort formula granules.

[0036] In a preferred embodiment of the present invention, if the test sample and the control medicinal material solution have fluorescent spots of the same color at the same Rf, and there is a fluorescent spot at Rf = 0.654, then it is the mugwort formula granules;

[0037] If there is no fluorescent spot at Rf=0.654, it is vinegar and mugwort formula granules.

[0038] The spotting amount of the thin layer chromatography test solution, vinegar wormwood formula granule solution and reference solution is 5 to 15 μL.

[0039] When 5-15 μl of the reference solution and the test solution are spotted, the spots are clearly colored and have good separation, and the spot volume is determined to be 5-15 μl.

[0040] The inspection temperature of the present invention is 4°C to 35°C.

[0041] This method has good durability at different temperatures. Compared with the reference sample spectrum, the test sample spectrum and the reference sample spectrum show spots of the same color at corresponding positions.

[0042] In some specific embodiments, at the ratio shift values of 0.634, 0.633, and 0.648, the mugwort formula particles show red fluorescent spots, while the vinegar mugwort formula particles have no spots.

[0043] This method has good durability to different humidity. Compared with the reference sample, the test sample and the reference sample show spots of the same color at corresponding positions.

[0044] In some specific embodiments, the mugwort formula particles show red fluorescent spots at the ratio shift values of 0.638, 0.610, and 0.667, while the vinegar mugwort formula particles have no spots.

[0045] The mugwort formula granules have red fluorescent spots at the above-mentioned ratio shift value (Rf), while the vinegar mugwort formula granules do not have such spots. The fluorescent spots can be used to distinguish the two.

[0046] The invention provides a thin layer chromatography identification method for mugwort and vinegar mugwort formula granules, comprising the following steps: A) pre-treating the mugwort formula granules to obtain a test solution; pre-treating the vinegar mugwort formula granules to obtain a vinegar mugwort formula granule solution; B) dissolving a scopolamine reference substance in methanol to obtain a reference substance solution; C) subjecting the test solution, the vinegar mugwort formula granule solution and the reference substance solution to thin layer chromatography, using a silica gel G thin layer plate and n-hexane-ethyl acetate-toluene as a developing agent; and D) inspecting under ultraviolet light, if the test solution and the reference medicinal material solution have fluorescent spots of the same color at the same Rf and fluorescent spots at Rf=0.625-0.741, the sample is the mugwort formula granule; if no fluorescent spots are present at Rf=0.625-0.741, the sample is the vinegar mugwort formula granule. The present invention discloses the construction and application of a thin-layer chromatography (TLC) identification method for mugwort and vinegar mugwort granules. By observing the TLC of a sample, the presence of a red fluorescent spot at a ratio shift value of 0.625 to 0.741 is used as an identification point for the mugwort granules, while the absence of a red fluorescent spot at the same location is used as an identification point for the vinegar mugwort granules. The established TLC method can quickly and effectively identify mugwort and vinegar mugwort granules. This method has strong specificity, good stability, high precision, and is convenient and easy to master. This method enables more standardized quality control of mugwort and vinegar mugwort granules, ensuring the efficacy of the drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 Spot sampling inspection;

[0048] Figure 2 Exclusive inspection;

[0049] Figure 3 For the investigation of different thin layer plates - Tianjin Silida;

[0050] Figure 4 Investigation of different thin layer plates - Merck;

[0051] Figure 5 Investigation of different thin layer plates-Qingdao Ocean;

[0052] Figure 6 Different temperatures -4°C;

[0053] Figure 7 Different temperatures -25°C;

[0054] Figure 8 Different temperatures 35°C;

[0055] Figure 9 Different humidity - 32%;

[0056] Figure 10 Different humidity - 51%

[0057] Figure 11 Different humidity - 75%;

[0058] Figure 12 Different times of verification;

[0059] Figure 13 This is the result diagram of the method of Comparative Example 1;

[0060] Figure 14 This is the result diagram of the method of Comparative Example 2;

[0061] Figure 15 This is the result diagram of comparative example 3 with the developing agent of n-hexane-ethyl acetate 9:1. DETAILED DESCRIPTION

[0062] To further illustrate the present invention, a thin layer identification method for mugwort and vinegar mugwort formula granules provided by the present invention is described in detail below in conjunction with examples.

[0063] Mortar, thin layer imaging system: CAMAG TLC Visualizer, silica gel G thin layer plate (Qingdao Ocean Chemical Plant, batch number: 20180527, Tianjin Silida Technology Co., Ltd., batch number: 191016, Merck, Germany, batch number: HX87183353)

[0064] Hexane, toluene, ethyl acetate, and methanol were all of analytical grade.

[0065] Ethanol, n-butanol, isopropanol, and glacial acetic acid were all analytical grade, and water was ultrapure water (prepared in the laboratory).

[0066] Scopoletin (China Food and Drug Inspection Institute, batch number: 110768-202105, purity 99.7%), mugwort formula granules (Sichuan New Green Pharmaceutical Technology Development Co., Ltd., batch numbers: 2208001; 2208002; 2208003) vinegar mugwort formula granules (Sichuan New Green Pharmaceutical Technology Development Co., Ltd., batch numbers: 2302081, 2302082, 2302083).

[0067] Example 1

[0068] Take 0.5g of this product, grind it finely, add 10ml of methanol and sonicate for 10 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 5ml of water, pass it through a D101 macroporous adsorption resin column (inner diameter 1cm, column height 14cm), elute with 20ml of 30% methanol, discard, and elute with 20ml of ethyl acetate. Collect the eluate, evaporate to dryness, and dissolve the residue in 1ml of ethyl acetate to prepare the test solution. For the reference solution of dongguan puer lactone, add methanol to prepare a solution containing 40μg per 1ml, which is used as the reference solution.

[0069] According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 Edition General Chapter 0502), a 10 μl spot of the test solution and the reference solution was aspirated and spotted on the same silica gel G thin layer plate. The plate was developed with n-hexane-ethyl acetate-toluene (1:3:2) as the developing solvent. The plate was removed, dried, and examined under ultraviolet light (365 nm). In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position in the chromatogram of the reference sample.

[0070] Example 2 Methodological Investigation

[0071] 2.1 Sampling quantity inspection

[0072] Under the experimental conditions proposed above, 5μl, 10μl and 15μl of the reference solution of dongguan lactone, the solution of mugwort formula granules and the solution of vinegar mugwort formula granules were taken and spotted on the same silica gel G thin layer plate. Figure 1 The results showed that when 5-15 μl of the reference and test solutions were spotted, the spots developed clearly and had good separation, indicating a suitable spot size of 5-15 μl. Furthermore, at a shift ratio of 0.674, the mugwort formula granules exhibited red fluorescent spots, while the vinegar mugwort formula granules showed no spots, which can be used to distinguish between the mugwort formula granules and the vinegar mugwort formula granules. The presence of a red fluorescent spot at a shift ratio of 0.674 can be used as a distinguishing factor.

[0073] Figure 1 Note: 1-3 are scopoletin reference solutions; 4-6 are mugwort granule solutions; 7-9 are vinegar mugwort granule solutions.

[0074] 2.2 Specificity Investigation

[0075] According to the above test sample preparation method, negative solution, scopoletin reference solution, mugwort formula granule solution, and vinegar mugwort formula granule solution were prepared respectively, and thin layer identification experiment was carried out. The results are shown in Figure 2 As can be seen from the figure, the negative sample did not interfere with the mugwort and vinegar mugwort granules, indicating that the method has good specificity. Furthermore, at the ratio shift value of 0.741, the mugwort granules showed red fluorescent spots, while the vinegar mugwort granules showed no spots. Figure 2 Specificity investigation. Note: 1 is the negative solution; 2 is the scopoletin reference solution; 3 is the mugwort granule solution; 4 is the vinegar mugwort granule solution.

[0076] 2.3 Durability Investigation

[0077] 2.3.1 Comparison of different thin layer plates

[0078] The tailorable thin layer chromatography plates of Tianjin Silida Technology Co., Ltd., Merck, and Qingdao Ocean Chemical Plant Branch prefabricated silica gel GF254 plates were selected and tested according to the proposed test methods. Figures 3-5The results showed that the red fluorescent spot ratio (Rf) values of the three thin-layer plates at 8.5 to 11 cm from the origin were 0.662, 0.659, and 0.655, respectively, all meeting the identification requirements. Furthermore, at Rf values of 0.613, 0.623, and 0.504, the mugwort formula granules showed red fluorescent spots, while the vinegar mugwort formula granules showed no spots.

[0079] The results are as follows Figure 3 , Figure 3 Note: 1 is scopoletin reference solution; 2 is mugwort granule solution; 3 is vinegar mugwort granule solution. Figure 4 Inspection of different thin layer plates - Merck Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution. Figure 5 Inspection of different thin layer plates - Qingdao Ocean, Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution.

[0080] 2.3.2 Comparison of different temperatures

[0081] Take the spotted thin layer plate and develop it at low temperature of 4℃, normal temperature of 25℃ and high temperature of 35℃ respectively. Figures 6-8 It can be seen that this method has good adaptability to different temperatures, and the mugwort formula particles show red fluorescent spots at the ratio shift values of 0.634, 0.633, and 0.648, while the vinegar mugwort formula particles have no spots. Figure 6 Different temperature 4℃; Note: Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution. Figure 7 Different temperatures: 25℃; Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution. Figure 8 Different temperature 35℃; Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution.

[0082] 2.3.3 Comparison of different humidity levels

[0083] Take the thin layer plate after spotting and develop it in the humidity environment of 32% rh, 51% rh and 75% rh respectively. Figures 9-11 .It can be seen from the figure that this method has good adaptability to different humidity, and the mugwort formula particles show red fluorescent spots at the ratio transfer values of 0.638, 0.610, and 0.667, while the vinegar mugwort formula particles have no spots. Figure 9 Different humidity - 32%, Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution; Figure 10Different humidity - 51%, Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution; Figure 11 Different humidity - 75%, Note: 1 is scopoletin reference solution; 2 is mugwort formula granule solution; 3 is vinegar mugwort formula granule solution.

[0084] 2.3.4 Verification

[0085] Scopoletin reference solution, Artemisia argyi formula granule solution, and Artemisia argyi vinegar formula granule solution were respectively spotted on the same silica gel GF254 thin layer plate and analyzed according to the determined method. The experimental results are shown in Figure 12 The results showed that the mugwort formula granules had a red fluorescent spot at a ratio shift value (Rf) of 0.625, while the vinegar mugwort formula granules did not have this spot. The fluorescent spot can be used to distinguish the two. Figure 12 Verification of different batches. Note: 1 is scopoletin reference solution; 2-4 are mugwort formula granule solutions; 5-7 are vinegar mugwort formula granule solutions.

[0086] 2.4 Determination of the Rf value of the identification point of mugwort formula granules and vinegar mugwort formula granules

[0087] The data of various ratio shift values in the thin layer chromatography methodology investigation are summarized and the results are shown in Table 1. Based on the summary results, it is determined that the ratio shift value (Rf) of the identification spot of the Artemisia argyi formula granules should be within the range of 15%, and the specified value is 0.654.

[0088] Table 1 Summary of methodological investigation ratios

[0089]

[0090] The results showed that at the ratio shift value of 0.654, there were no fluorescent spots in the vinegar and mugwort formula granules, while the other mugwort formula granules had obvious red fluorescent spots, which could be used as an identification point to distinguish between the mugwort formula granules and the vinegar and mugwort formula granules.

[0091] Comparative Example 1

[0092] Take 4g of this product, add 10mL of chloroform, ultrasonicate for 15min, and then let it stand for 1h. Filter the filtrate to dryness and dissolve it in 1mL of chloroform as the test solution. Take another reference substance of dongguanpollactone, add methanol to make a solution containing 40μg per 1ml, as the reference solution. Take 4μL of the above solution. Spot them on the same silica gel G thin layer plate, develop them with cyclohexane-ether-ethyl acetate (20:5.5:2.5) as the developing agent, take them out to dry, spray them with 10% sulfuric acid ethanol solution, and bake them at 100℃ for about 10min to develop color. In the chromatogram of the test product, fluorescent spots of the same color appear at the corresponding position of the chromatogram of the reference substance. See the experimental results. Figure 13 . Figure 13The figure shows the results of the method of Comparative Example 1. Note: 1 is the scopoletin reference solution; 2-4 are the mugwort formula granule solutions; 5-8 are the vinegar mugwort formula granule solutions.

[0093] The results showed that using the above-mentioned thin layer identification method, there were no obvious spots on the mugwort leaves and the vinegar mugwort formula granules, and the two could not be identified.

[0094] Comparative Example 2

[0095] Grind 1g of this product, add 30ml of 70% ethanol and 2ml of hydrochloric acid, heat under reflux for 1 hour, filter, and dry the filtrate. Dissolve the residue in 20ml of water. Shake and extract twice with 20ml of acid and ester. Combine the ethyl acetate solutions, evaporate to dryness, and dissolve the residue in 1ml of methanol to prepare the test solution. Separately, take 3g of Artemisia argyi as a control medicinal material, add 50ml of water, boil for 30 minutes, filter, and evaporate the filtrate to dryness. Add 30ml of 70% ethanol and 2ml of hydrochloric acid to the residue and prepare a control medicinal material solution in the same manner. Also, take the dongliang lactone reference material and add methanol to a solution containing 40μg per 1ml. This is used as the reference solution. According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 edition General Rules 0502), 5ul of each of the above solutions were taken and spotted on the same silica gel G thin layer plate, and chloroformane-methanol-formic acid (20:3.5:2) was used as the developing agent. The plate was developed, taken out, dried, sprayed with aluminum chloride test solution, dried with hot air, and inspected under ultraviolet light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding positions of the chromatograms of the reference sample and the reference medicinal material. The experimental results are shown in Figure 14 shown. Figure 14 The figure shows the results of the method of comparative example 2. Note: 1 is the scopoletin reference solution; 2 is the wormwood reference medicinal material solution; 3 is the wormwood formula granule solution; 4 is the vinegar wormwood formula granule solution.

[0096] The results showed that using the above-mentioned thin layer identification method, there were no obvious different spots between mugwort leaves and vinegar mugwort formula granules, and the two could not be distinguished.

[0097] Comparative Example 3

[0098] Take 1g of this product, grind it into powder, add 50ml of ether, ultrasonicate for 30min, filter, evaporate the filtrate, and dissolve the residue in 1ml of ethyl acetate to prepare the test solution. According to the thin layer chromatography method (General Method 0502), take 10μl of the above solution and spot it on a silica gel G thin layer plate. Use n-hexane-ethyl acetate (9:1) as the developing solvent, develop, remove, dry, use 10% sulfuric acid ethanol solution as the color developing agent, heat at 105℃, and examine under ultraviolet light (365nm). The experimental results are shown in Figure 15 shown. Figure 15This is the result of developing with a 9:1 ratio of n-hexane to ethyl acetate. Note: 1 is the scopoletin reference solution; 2 is the wormwood granule solution; 3 is the vinegar wormwood granule solution.

[0099] The results showed that using the above-mentioned thin layer identification method, there were no obvious different spots between mugwort leaves and vinegar mugwort formula granules, and the two could not be distinguished.

[0100] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A thin layer identification method for mugwort and mugwort vinegar granules, comprising: A) Pre-treating the Artemisia argyi granules to obtain a test solution; the pre-treating of the Artemisia argyi granules or vinegar Artemisia argyi granules is specifically as follows: a) adding methanol to the granules of Artemisia argyi formula or the granules of Artemisia argyi formula with vinegar, sonicating, filtering, evaporating, dissolving the residue in water, passing it through a D101 macroporous adsorption resin column, eluting with 30% methanol, discarding the eluate, and then eluting with ethyl acetate, collecting the eluate; b) Recover the solvent from the eluate until it is dry, and dissolve the residue in ethyl acetate to obtain the product; Pre-treating the vinegar wormwood formula granules to obtain a vinegar wormwood formula granule solution; B) Take the scopolamine reference substance and dissolve it in methanol to obtain a reference solution; C) subjecting the test solution, vinegar wormwood granule solution, and reference solution to thin layer chromatography using a silica gel G thin layer plate and a developing solvent of n-hexane-ethyl acetate-toluene in a mass ratio of 1:3:2; D) Observe under ultraviolet light. If the test and control solutions have fluorescent spots of the same color at the same Rf, and there are fluorescent spots at Rf = 0.625-0.741, then it is the mugwort formula granules. If there is no fluorescent spot at Rf=0.625~0.741, it is vinegar and mugwort formula granules.

2. The method according to claim 1, characterized in that Step a) the ultrasonic time is 5 to 10min; the formula of wormwood granules or vinegar wormwood granules, methanol, water, 30% methanol and ethyl acetate mass volume ratio of 0.5g: 10mL: 5mL: 20mL: 20mL; The D101 macroporous adsorption resin has an inner diameter of 1 cm and a column height of 14 cm.

3. The method according to claim 1, characterized in that In step B), the mass volume ratio of the scopolamine reference substance and methanol is 40 μg:1 mL.

4. The method according to claim 1, wherein The sample spotting volume of the thin layer chromatography test solution, vinegar wormwood formula granule solution and reference solution is 5-15 μL.

5. The method according to claim 1, wherein In step D), the wavelength of the ultraviolet lamp is 365 nm.

6. The method according to claim 1, characterized in that Step D) The silica gel G thin layer plate is a GF254 thin layer plate.

7. The method according to claim 6, characterized in that Step D) checking the temperature to be 4°C to 35°C; checking the humidity to be 32% RH to 75% RH.

8. The method according to claim 1, characterized in that The Rf value in step D) is 0.654.

Images