Detection method of medicine for dispelling wind and eliminating dampness

Abstract

The invention relates to a detection method of a medicine for dispelling wind and eliminating dampness. The detection method comprises the following steps: thin-layer identification of a notopterygium root medicinal material, thin-layer identification of a radix angelicae pubescentis medicinal material, thin-layer identification of a radix saposhnikoviae medicinal material, thin-layer identification of ligusticum and ligusticum wallichii medicinal materials, and thin-layer identification of a honey-fried licorice root medicinal material. Through methodological investigation, the detection method has the effects of strong specificity, good durability, clear spots after thin layer expansion, satisfactory separation degree and the like, can effectively control the product quality condition, improves the product quality detection standard, and ensures the product quality.

Description

Technical Field

[0001] This invention relates to the field of drug detection, specifically to a method for detecting drugs that dispel wind and dampness. Background Technology

[0002] Qianghuo Shengshi Decoction is composed of seven Chinese herbs: Qianghuo (Notopterygium root), Duhuo (Angelica pubescens root), Gaoben (Ligusticum striatum root), Fangfeng (Saposhnikovia root), Gancao (Glycyrrhiza root), Manjingzi (Vitex trifolia fruit), and Chuanxiong (Ligusticum striatum rhizome). It has the effects of dispelling wind, eliminating dampness, and relieving pain. Clinically, it is often used to treat rheumatoid arthritis, osteoarthritis, osteoproliferative disorders, ankylosing spondylitis, cervical spondylosis, and shoulder, back, and headache.

[0003] Currently, most quality testing methods for Qianghuo Shengshi Decoction focus on controlling fingerprint spectroscopy and content determination, which cannot effectively control the quality of the decoction.

[0004] CN117233279A discloses a method for quality detection of Qianghuo Shengshi Decoction composition and its application. This invention belongs to the field of pharmaceutical component detection technology and provides a method for quality detection of Qianghuo Shengshi Decoction composition and its application, including: dissolving a reference material in methanol to obtain a reference solution; dissolving the Qianghuo Shengshi Decoction composition in methanol solution, sonicating, filtering the supernatant to obtain a test solution; and injecting the obtained reference solution and the obtained test solution into a liquid chromatograph for determination. The method for quality detection of Qianghuo Shengshi Decoction composition provided by this invention detects the test solution and the reference solution under the same chromatographic conditions, obtaining chromatograms of the test solution and the reference solution. The measured ammonium glycyrrhizate and osthol show a good linear correlation with the peak area. It is also convenient, rapid, specific, stable, accurate, reproducible, system suitability, intermediate precision, and robustness, and can be effectively applied to Qianghuo Shengshi Decoction composition.

[0005] Determination of the content of Angelica pubescens and Glycyrrhiza uralensis in the sample.

[0006] CN113219076A discloses a method for preparing a standard decoction of Qianghuo Shengshi Decoction and its fingerprint spectrum. This invention belongs to the field of traditional Chinese medicine compound prescription technology. The preparation of the standard decoction includes the following steps: taking each medicinal material in the prescription, pulverizing them separately, mixing them in proportion, adding distilled water, soaking for half an hour, first boiling over high heat, then simmering over low heat, filtering while hot after simmering, and centrifuging to obtain the final product. The standard decoction preparation method provided by this invention can achieve a high yield of extract and a high extraction rate of active ingredients. The method for establishing the fingerprint spectrum includes the following steps: preparation of different batches of test solution, preparation of reference solution, determination and construction of fingerprint spectrum. The fingerprint spectrum establishment method provided by this invention has high stability, repeatability and precision, and is simple and easy to implement. The established fingerprint spectrum has 20 common peaks, including cimicifugain glycoside, ferulic acid, glycyrrhizic acid ammonium, vitexin, notopterygol, osthol and isoimperatorin, which improves the quality control method of Qianghuo Shengshi Decoction.

[0007] To address the above issues, better ensure the quality of Qianghuo Shengshi Decoction, and improve drug quality standards, the inventors, based on the company's actual situation and in accordance with the Chinese Pharmacopoeia, developed thin-layer chromatography (TLC) identification tests for Qianghuo Shengshi Decoction. These tests include TLC identification of Qianghuo, Duhuo, Fangfeng, Gaoben, Chuanxiong, and Zhigancao, which more effectively control product quality and thus guarantee clinical efficacy. Summary of the Invention

[0008] The purpose of this invention is to provide a method for detecting the drug Qianghuo Shengshi Decoction.

[0009] The prescription and preparation method of the drug described in this invention are as follows:

[0010] Prescription: Notopterygium root 4.13g, Ligusticum root 2.06g, Prepared licorice root 2.06g, Angelica pubescens root 4.13g, Saposhnikovia root 2.06g

[0011] Chuanxiong (Ligusticum striatum) 2.06g, Manjingzi (Vitex trifolia) 1.24g

[0012] Preparation: Crush the above seven ingredients into coarse particles. Weigh each crushed ingredient according to the single-dose dosage, place them in a clay pot, add 400ml of water, bring to a boil over high heat, simmer over low heat for 31 minutes, filter, concentrate the filtrate under reduced pressure until the ratio of the amount of medicinal slices to the weight of the concentrate is 1:2 to 1:4, freeze dry to obtain dry powder, package, and obtain the corresponding physical product.

[0013] The detection methods for the drugs described in this invention are as follows: thin-layer chromatography identification of Notopterygium incisum; thin-layer chromatography identification of Angelica pubescens; thin-layer chromatography identification of Saposhnikovia divaricata; thin-layer chromatography identification of Ligusticum striatum and Ligusticum chuanxiong; and thin-layer chromatography identification of Glycyrrhiza uralensis (processed).

[0014] The thin-layer chromatography identification method for Notopterygium incisum described in this invention is as follows: Take 0.5-1.5g of the corresponding drug sample, add 25-35ml of methanol, sonicate for 20-40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-3ml of methanol to obtain the test solution; separately take 0.5-1.5g of Notopterygium incisum reference material, add 40-60ml of water and decoct for 25-35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; take 1μl each of the test solution and the reference material solution and spot them separately on the same silica gel G thin-layer plate, using a 3:1:2 ratio of n-hexane-60-90℃ petroleum ether-ethyl acetate as the developing solvent, develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference material.

[0015] Preferably, the thin-layer chromatography identification method for Notopterygium incisum described in this invention is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution; Take 1g of Notopterygium incisum reference material, add 50ml of water and decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Take 1μl each of the test solution and the reference material solution and spot them separately on the same silica gel G thin-layer plate, using a 3:1:2 ratio of n-hexane-60~90℃ petroleum ether-ethyl acetate as the developing solvent, develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference material.

[0016] The thin-layer chromatography identification method for Angelica pubescens described in this invention is as follows: Take 0.5-1.5g of the corresponding drug sample, add 25-35ml of methanol, sonicate for 20-40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-3ml of methanol to prepare the test solution; Take 0.5-1.5g of Angelica pubescens reference material, add 40-60ml of water, decoct for 25-35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Apply 5μl each of the test solution and the reference material solution to the same silica gel GF plate according to the thin-layer chromatography method. 254 On a thin-layer plate, dichloromethane-ethyl acetate in a ratio of 7:4 was used as the developing solvent. The plate was then removed, dried, and sprayed with 10% sulfuric acid ethanol solution. The plate was heated at 105°C until the spots were clearly visible. The plate was then examined under a UV lamp at 254 nm. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position as in the chromatogram of the reference medicinal material.

[0017] Preferably, the thin-layer chromatography identification method for Angelica pubescens described in this invention is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution; Take 1g of Angelica pubescens reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Apply 5μl each of the test solution and the reference material solution to the same silica gel GF plate according to the thin-layer chromatography method. 254 On a thin-layer plate, dichloromethane-ethyl acetate in a ratio of 7:4 was used as the developing solvent. The plate was then removed, dried, and sprayed with 10% sulfuric acid ethanol solution. The plate was heated at 105°C until the spots were clearly visible. The plate was then examined under a UV lamp at 254 nm. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position as in the chromatogram of the reference medicinal material.

[0018] The thin-layer chromatography identification method for Saposhnikovia divaricata described in this invention is as follows: Take 1-3g of the corresponding drug sample, add 10-30ml of acetone, sonicate for 10-30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.4-0.6ml of ethanol to prepare the test solution; Separately, take 0.1-0.3g of Saposhnikovia divaricata reference material, add 40-60ml of water, decoct for 25-35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Then, take cimicifugoside reference standard and 5-O-methylvisamidolol reference standard, add ethanol... Prepare a mixed solution containing 1 mg of each of the test sample and the reference material solution per 1 ml as a reference solution. Using thin-layer chromatography, apply 6 μl of the test sample solution, 3 μl of the reference material solution, and 1 μl of the reference solution separately to the same silica gel G thin-layer plate. Develop the plate using a 4:1 dichloromethane-methanol mixture as the developing solvent. Remove the plate, air-dry it, spray it with 10% sulfuric acid ethanol solution, air-dry it again, and examine it under a 365 nm UV lamp. In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference solution.

[0019] Preferably, the thin-layer chromatography identification method for Saposhnikovia divaricata of the present invention is as follows: Take 2g of the corresponding drug sample, add 20ml of acetone, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.5ml of ethanol to obtain the test solution; separately take 0.2g of Saposhnikovia divaricata reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; then take cimicifugoside reference standard and 5-O-methylvisamidolol reference standard, add ethanol to prepare a solution containing 1g of each per ml. A mixed solution of mg was used as the reference solution. 6 μl of the test solution, 3 μl of the reference medicinal material solution, and 1 μl of the reference solution were spotted separately onto the same silica gel G thin-layer plate using a 4:1 dichloromethane-methanol mixture as the developing solvent. The plate was then removed, dried, sprayed with 10% sulfuric acid ethanol solution, dried again, and examined under a 365 nm UV lamp. Fluorescent spots of the same color appeared at the corresponding positions in the chromatograms of the test sample, the reference medicinal material, and the reference solution.

[0020] The thin-layer chromatography method for identifying Ligusticum striatum and Ligusticum chuanxiong described in this invention is as follows: Take 0.5-1.5g of the corresponding drug sample, add 25-35ml of methanol, sonicate for 20-40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-3ml of methanol to obtain the test solution; take 0.1-0.3g each of Ligusticum striatum and Ligusticum chuanxiong reference materials to prepare the Ligusticum striatum and Ligusticum chuanxiong reference material solutions using the same method as the test solution; separately take ligustilide reference standard, add methanol to prepare a solution containing 1mg per ml to obtain the reference solution; according to the thin-layer chromatography method, take 5μl each of the test solution, reference material solution, and reference solution, and spot them separately on the same silica gel G thin-layer plate, using a 5:1 ratio of petroleum ether-ethyl acetate at 60-90℃ as the developing solvent, develop, remove, air dry, and examine under a UV lamp at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference materials and the reference solution.

[0021] Preferably, the thin-layer chromatography identification method for Ligusticum striatum and Ligusticum chuanxiong of the present invention is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution; take 0.2g each of Ligusticum striatum and Ligusticum chuanxiong reference materials to prepare Ligusticum striatum and Ligusticum chuanxiong reference material solutions in the same way as the test solution; separately take ligustilide reference standard, add methanol to prepare a solution containing 1mg per 1ml to obtain the reference solution; according to the thin-layer chromatography method, take 5μl each of the test solution, reference material solution, and reference solution, and spot them separately on the same silica gel G thin-layer plate, use petroleum ether-ethyl acetate at a ratio of 5:1 at 60-90℃ as the developing solvent, develop, remove, air dry, and examine under a UV lamp at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference materials and the reference solution.

[0022] The thin-layer chromatography method for identifying processed licorice root according to this invention is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution; Take 0.2g of licorice root reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Separately, take glycyrrhizin reference standard, add methanol to prepare a solution containing 1mg per ml to obtain the reference standard solution; Apply 2μl each of the test solution, reference material solution, and reference standard solution to the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution according to the thin-layer chromatography method, develop the plate using ethyl acetate-formic acid-glacial acetic acid-water in a ratio of 15:1:1:2, remove the plate, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible; In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference standard.

[0023] Beneficial effects

[0024] 1. The detection method of this invention includes thin-layer chromatography (TLC) identification of the following medicinal materials: Notopterygium incisum, Angelica pubescens, Saposhnikovia divaricata, Ligusticum striatum, Ligusticum chuanxiong, and processed licorice root. Through methodological investigation, this detection method demonstrates strong specificity, good durability, clear spots after TLC development, and satisfactory separation, effectively controlling product quality, improving product quality testing standards, and ensuring product quality.

[0025] 2. The thin-layer chromatography identification method for Notopterygium incisum of this invention was first tested with reference to the thin-layer chromatography examination method under the "Notopterygium incisum" section of the 2015 edition of the Chinese Pharmacopoeia. The results showed that the negative sample lacking Notopterygium incisum interfered with the corresponding position of the purpureusin reference material. Further research was conducted by changing the developing system and viewing conditions. The results showed that the spots in the chromatogram of the test sample had good separation, with the same main spot color as the Notopterygium incisum reference material at the corresponding position, and the Rf value was moderate. The negative sample lacking Notopterygium incisum did not interfere, thus establishing the thin-layer chromatography identification method for Notopterygium incisum. The method underwent robustness testing, examining the effects of different manufacturers' silica gel G thin-layer plates (Yantai Jiangyou Silica Gel Development Co., Ltd. and Qingdao Ocean Chemical Plant), different developing temperatures, and different relative humidity. The results showed good robustness under all developing conditions, with clear main spots. Fifteen batches of samples were tested, and the results showed that the test sample chromatogram showed the same main spot color as the reference material chromatogram at the corresponding position. The methodological test results indicate that the method is simple, accurate, stable, and reproducible; therefore, it was included in the quality standard text.

[0026] 3. The thin-layer chromatography identification method for Angelica pubescens of this invention was first tested with reference to the thin-layer chromatography examination method under the "Angelica pubescens" section of the 2015 edition of the Chinese Pharmacopoeia. The results showed that in the chromatogram of the test sample, the fluorescent spot at the corresponding position to the osthol reference standard was lighter; and the spot at the corresponding position to the dihydroapigenin angelica ester reference standard was missing. Interference existed in the negative sample, so the developing system of Notopterygium incisum was further analyzed. The results showed that under ultraviolet light (365nm), the chromatogram of the test sample showed fluorescent spots of the same color at the corresponding positions as the chromatogram of Angelica pubescens slices, but the negative sample showed interference, making the method infeasible. Changing the viewing conditions to ultraviolet light (254nm), the chromatogram of the test sample showed fluorescent spots of the same color at the corresponding positions as the reference herb chromatogram, and the negative sample showed no interference, with a moderate Rf value.

[0027] The method is feasible, therefore a thin-layer chromatography method for identifying Angelica pubescens was determined. A robustness test was conducted on this method, examining the effects of different manufacturers' silica gel G thin-layer plates (Yantai Jiangyou Silica Gel Development Co., Ltd. and Qingdao Ocean Chemical Plant), different developing temperatures, and different relative humidity levels. The results showed good robustness under all developing conditions, with clear main spots. Fifteen batches of samples were tested, and the results showed that the test sample chromatogram showed spots of the same color at the corresponding positions as the reference herb chromatogram. The methodological test results indicate that the method is simple, accurate, stable, and reproducible; therefore, it was included in the quality standard text.

[0028] 4. The thin-layer chromatography identification method for Saposhnikovia divaricata of this invention first refers to the thin-layer chromatography test method under the "Saposhnikovia divaricata" section of the 2015 edition of the Chinese Pharmacopoeia. The results showed that the chromatogram of the test sample had a heavy band, and the chromatogram of the test sample was almost identical to the negative chromatogram lacking Saposhnikovia divaricata, indicating interference from the negative chromatogram. Therefore, further research was conducted on the developing solvent system and colorimetric conditions. The results showed that in the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding positions as in the chromatograms of the reference medicinal material and the reference standard, with no interference from the negative chromatogram and a moderate Rf value, indicating the method's feasibility. Therefore, the thin-layer chromatography identification method for Saposhnikovia divaricata was determined. The method underwent a durability test, examining the effects of different manufacturers' silica gel G thin-layer plates (Yantai Jiangyou Silica Gel Development Co., Ltd. and Qingdao Ocean Chemical Plant), different developing temperatures, and different relative humidity. The results showed good durability under various developing conditions, with clear fluorescent spots. Fifteen batches of samples were tested, and the results showed that in the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding positions as in the chromatograms of the reference medicinal material and the reference standard. The methodological test results show that the method is convenient, accurate, stable, and has good reproducibility, therefore it is included in the main text of the quality standard.

[0029] 5. Both Ligusticum striatum and Ligusticum chuanxiong of this invention contain indicative components such as ferulic acid and ligustilide. Therefore, the two herbs were combined for thin-layer chromatography (TLC) exploration. The TLC identification method was first referenced from the TLC method under the "Identification" section of the 2015 edition of the Chinese Pharmacopoeia, Part I. The results showed that Ligusticum chuanxiong, Ligusticum striatum, and Notopterygium incisum all contained ferulic acid. This component is not specific, therefore ferulic acid was not included in the standard. The chromatogram of the test sample showed fluorescent spots of the same color at the corresponding positions of ligustilide, and the absence of double negatives from Ligusticum striatum and Ligusticum chuanxiong resulted in no interference, but the Rf value was high. After adjusting the developing system, the chromatogram of the test sample showed fluorescent spots of the same color at the corresponding positions as the chromatograms of the reference herbs and the reference substance, with no interference from negatives and a moderate Rf value. Therefore, the method is feasible, and the TLC identification method for Ligusticum striatum and Ligusticum chuanxiong was determined. This method underwent durability testing to investigate the effects of different manufacturers' silica gel G thin-layer plates (Yantai Jiangyou Silica Gel Development Co., Ltd. and Qingdao Ocean Chemical Plant), as well as different developing temperatures and relative humidity. The results showed good durability under all developing conditions, with clear fluorescent spots. Fifteen batches of samples were tested, and the results showed fluorescent spots of the same color at the corresponding positions as in the chromatograms of the reference medicinal material and the reference standard. The methodological results demonstrate that the method is simple, accurate, stable, and reproducible; therefore, it was included in the quality standard text.

[0030] 6. The thin-layer chromatography identification method for prepared licorice root of this invention first refers to the thin-layer chromatography examination method under the "Licorice" section of the 2015 edition of the Chinese Pharmacopoeia. The results showed that under sunlight, the chromatogram of the test sample showed spots of the same color at the corresponding positions as the reference standard and licorice root slices, with no negative interference, but the spots on the test sample were lighter in color. Under ultraviolet light (365nm), the chromatogram of the test sample showed no corresponding spots as the reference standard and lyophilized licorice root powder. Therefore, the preparation method of the test sample was optimized and simplified. The test sample obtained by ultrasonication with methanol was then used for the experiment. Under sunlight, the chromatogram of the test sample showed four identical yellow spots at the corresponding positions as the chromatogram of the licorice reference material, with no negative interference. Under ultraviolet light (365nm), the chromatogram of the test sample showed mainly three identical fluorescent spots at the corresponding positions as the chromatogram of the licorice reference material, with no negative interference; the remaining fluorescent spots were negative and interfered with. This indicates that the effective chromatographic information under ultraviolet light (365nm) is less than that under sunlight, and the chromatographic information under the two viewing conditions is not complementary. Therefore, sunlight was determined as the viewing condition, and the thin-layer chromatography method for identifying processed licorice was established. The method underwent a robustness test, examining the effects of different manufacturers' 1% sodium hydroxide silica gel G thin-layer plates (Yantai Jiangyou Silica Gel Development Co., Ltd. and Qingdao Ocean Chemical Plant), different developing temperatures, and different relative humidity. The results showed good robustness under all developing conditions, with clear spots. Fifteen batches of samples were tested, and the results showed that the chromatogram of the test sample showed spots of the same color at the corresponding positions as the chromatograms of the reference material and the reference substance. Attached Figure Description

[0031] Figure 1 Thin-layer chromatogram of Notopterygium incisum - Method 1 (1 in the figure is purpureusin reference standard; 2 is Notopterygium incisum slices; 3 is notopterygium incisum slices; 4 is notopterygium incisum slices; 5 is notopterygium incisum slices; 6 is notopterygium incisum slices; 7 is notopterygium incisum slices; 8 is notopterygium

[0032] 4 is the test sample; 4 is a negative sample lacking Notopterygium incisum.

[0033] Figure 2 Thin-layer chromatogram of Notopterygium incisum - Method 2 (1 in the figure is Notopterygium incisum reference material; 2-4 are all test samples; 5 is a negative sample lacking Notopterygium incisum).

[0034] Figure 3 Durability test of Notopterygium incisum thin-layer plate (Yantai silica gel G thin-layer plate, T 19.4℃, RH 13%); Figure 1

[0035] 1 is the reference material for Notopterygium incisum; 2-4 are all test samples; 5 is a negative sample lacking Notopterygium incisum.

[0036] Figure 4 Durability test of Notopterygium incisum thin-layer chromatography (Qingdao silica gel G thin-layer plate, T 19.1℃, RH 12%); Figure 1

[0037] 1 is the reference material for Notopterygium incisum; 2-4 are all test samples; 5 is a negative sample lacking Notopterygium incisum.

[0038] Figure 5 Durability test of Notopterygium incisum thin-layer plate (Yantai silica gel G thin-layer plate, T 21.0℃, RH 77%); Figure 1

[0039] 1 is the reference material for Notopterygium incisum; 2-4 are all test samples; 5 is a negative sample lacking Notopterygium incisum.

[0040] Figure 6 Durability test of Notopterygium incisum thin-layer plate (Yantai silica gel G thin-layer plate, T 2.9℃, RH 35%); Figure 1

[0041] 1 is the reference material for Notopterygium incisum; 2-4 are all test samples; 5 is a negative sample lacking Notopterygium incisum.

[0042] Figure 7 Thin-layer chromatography (TLC) images of Notopterygium incisum in 15 batches of samples (Figure 1 is Notopterygium incisum reference material; 2-16 are test samples, batch numbers are 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, 181204-15).

[0043] Figure 8Thin-layer chromatography (TLC) images of Notopterygium incisum in 15 batches of samples (17 in the figure is Notopterygium incisum reference material; 18-32 are the test samples, batch numbers 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, 181207-30).

[0044] Figure 9 Thin-layer chromatogram of Angelica pubescens - Method 1 (In the figure, 1 is dihydroaperitol angelica ester reference standard; 2 is Angelica pubescens slices; 3 is osthol reference standard; 4 is negative sample lacking Angelica pubescens; 5-7 are all test samples);

[0045] Figure 10 Thin-layer chromatogram of Angelica pubescens - Method 2 (1 and 2 in the figure are Angelica pubescens slices; 3 and 4 are test samples; 5 and 6 are negative samples lacking Angelica pubescens);

[0046] Figure 11 Thin-layer chromatogram of Angelica pubescens - Method 3 (1 in the figure is Angelica pubescens reference material; 2-4 are all test samples; 5 is a negative sample lacking Angelica pubescens);

[0047] Figure 12 Thin-layer chromatography of Angelica pubescens durability (Yantai Silica Gel GF) 254 Thin-layer plate, T 24.3℃, RH 11%; 1 in the figure is Angelica pubescens control material; 2-4 are test samples; 5 is a negative sample lacking Angelica pubescens.

[0048] Figure 13 Thin-layer chromatography (TLC) of Angelica pubescens durability test (Qingdao Silica Gel GF) 254 Thin-layer plate, T 24.3℃, RH 11%; 1 in the figure is Angelica pubescens control material; 2-4 are test samples; 5 is a negative sample lacking Angelica pubescens.

[0049] Figure 14 Thin-layer chromatography (TLC) of Angelica pubescens durability test (Qingdao Silica Gel GF) 254 Thin-layer plate, T 20.1℃, RH 80%; 1 in the figure is Angelica pubescens control material; 2-4 are test samples; 5 is a negative sample lacking Angelica pubescens.

[0050] Figure 15 Thin-layer chromatography (TLC) of Angelica pubescens durability test (Qingdao Silica Gel GF) 254 Thin-layer plate, T 2.0℃, RH 80%; 1 in the figure is Angelica pubescens control material; 2-4 are test samples; 5 is a negative sample lacking Angelica pubescens.

[0051] Figure 16Thin-layer chromatography (TLC) images of Angelica pubescens from 15 batches of samples (Figure 1 shows the control material of Angelica pubescens; 2-16 are batch numbers 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, and 181204-15, respectively).

[0052] Figure 17 Thin-layer chromatography (TLC) images of Angelica pubescens from 15 batches of samples (Figure 17 shows the control material; 18-32 represent batches 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, and 181207-27, respectively).

[0053] 181207-28, 181207-29, 181207-30);

[0054] Figure 18 Thin-layer chromatogram of Saposhnikovia divaricata - Method 1 (In the figure, 1 is 5-O-methylvisamidol glycoside reference standard; 2 is Saposhnikovia divaricata slices; 3 is cimicifugoside reference standard; 4 is negative sample without Saposhnikovia divaricata; 5 is test sample);

[0055] Figure 19 Thin-layer chromatogram of Saposhnikovia divaricata - Method 2 (In the figure, 1-3 are 1 μl, 2 μl, and 3 μl of cimicifugain glycoside reference standard, respectively; 4-6 are 1 μl, 2 μl, and 3 μl of 5-O-methylvisamidol glycoside reference standard, respectively; 7-9 are 1 μl, 2 μl, and 3 μl of Saposhnikovia divaricata reference material, respectively; 10-12 are the test materials, respectively).

[0056] 5μl, 6μl, and 7μl of sample; 13 is a negative sample solution lacking Saposhnikovia divaricata.

[0057] Figure 20 Thin-layer chromatography diagram for the durability test of Saposhnikovia divaricata (Yantai silica gel G thin-layer plate, T 14.0℃, RH 29.8%; 1 in the figure is cimicifugoside reference standard; 2 is 5-O-methylvisamilol glycoside reference standard; 3 is Saposhnikovia divaricata reference medicinal material; 4-6 are all test samples; 7 is negative sample without Saposhnikovia divaricata).

[0058] Figure 21Thin-layer chromatography diagram for the durability test of Saposhnikovia divaricata (Qingdao silica gel G thin-layer plate, T 14.0℃, RH 29.8%; 1 in the figure is cimicifugoside reference standard; 2 is 5-O-methylvisamidol glycoside reference standard; 3 is Saposhnikovia divaricata reference medicinal material; 4-6 are all test samples; 7 is negative sample without Saposhnikovia divaricata).

[0059] Figure 22 Thin-layer chromatography diagram for the durability test of Saposhnikovia divaricata (Qingdao silica gel G thin-layer plate, T 17.2℃, RH 80%; 1 in the figure is the reference standard of cimicifuga glycoside; 2 is the reference standard of 5-O-methylvisammiol glycoside; 3 is the reference medicinal material of Saposhnikovia divaricata; 4-6 are all test samples; 7 is the negative sample without Saposhnikovia divaricata).

[0060] Figure 23 Thin-layer chromatography diagram for the durability test of Saposhnikovia divaricata (Qingdao silica gel G thin-layer plate, T 2.0℃, RH 80%; 1 in the figure is cimicifugoside reference standard; 2 is 5-O-methylvisamidol glycoside reference standard; 3 is Saposhnikovia divaricata reference medicinal material; 4-6 are all test samples; 7 is negative sample without Saposhnikovia divaricata).

[0061] Figure 24 Thin-layer chromatography (TLC) images of Saposhnikovia divaricata samples from 15 batches (Figure 1: cimicifugoside reference standard; 2: cimicifugoside reference standard; 3: Saposhnikovia divaricata reference material; 4-18: batch numbers 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, 181204-15).

[0062] Figure 25 Thin-layer chromatography (TLC) images of Saposhnikovia divaricata samples from 15 batches (Figure 19 is cimicifugoside reference standard; 20 is cimicifugoside reference standard; 21 is Saposhnikovia divaricata reference medicinal material; 22-36 are batch numbers 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, 181207-30, respectively).

[0063] Figure 26 Thin-layer chromatograms of Ligusticum striatum and Ligusticum chuanxiong - Method 1 (In the figure, 1 is Ligusticum chuanxiong slices; 2 is Ligusticum striatum slices; 3 is Notopterygium incisum slices; 4 is ferulic acid reference standard; 5 is ligustilide reference standard; 6 is the test sample; 7 is a double negative sample lacking both Ligusticum chuanxiong and Ligusticum striatum; 8 is a negative sample lacking Ligusticum chuanxiong; 9 is a negative sample lacking Ligusticum striatum).

[0064] Figure 27 Thin-layer chromatograms of Ligusticum striatum and Ligusticum chuanxiong - Method 2 (Figures 1-3 are ligustilide reference standard 2 in order)

[0065] μl, 3μl, 5μl; 4-6 are 2μl, 3μl, 5μl of Ligusticum chuanxiong reference material, respectively; 7-9 are 2μl, 3μl, 5μl of Ligusticum striatum reference material, respectively; 10-12 are 2μl, 3μl, 5μl of test sample, respectively; 13 is a negative sample lacking Ligusticum chuanxiong; 14 is a negative sample lacking Ligusticum striatum; 15 is a double negative sample lacking both Ligusticum chuanxiong and Ligusticum striatum.

[0066] Figure 28 Thin-layer chromatography (TLC) results for durability testing of Ligusticum striatum and Ligusticum chuanxiong (Yantai silica gel G TLC plate, T 15.6℃, RH 29.8%); 1 in the figure is ligustilide reference standard; 2 is Ligusticum chuanxiong reference material; 3 is Ligusticum striatum reference material; 4-5 are all test samples; 6 is negative sample lacking Ligusticum chuanxiong; 7 is negative sample lacking Ligusticum striatum; 8 is double negative sample lacking both Ligusticum chuanxiong and Ligusticum striatum).

[0067] Figure 29 Thin-layer chromatography (TLC) results for durability testing of Ligusticum striatum and Ligusticum chuanxiong (Qingdao silica gel G TLC plate, T 15.6℃, RH 29.8%); 1 in the figure is ligustilide reference standard; 2 is Ligusticum chuanxiong reference material; 3 is Ligusticum striatum reference material; 4-5 are all test samples; 6 is negative sample lacking Ligusticum chuanxiong; 7 is negative sample lacking Ligusticum striatum; 8 is double negative sample lacking both Ligusticum chuanxiong and Ligusticum striatum).

[0068] Figure 30 Durability test of Ligusticum striatum and Ligusticum chuanxiong (Qingdao silica gel G thin film, T 17.3℃, RH 79%)

[0069] In the figure, 1 is ligustilide reference standard; 2 is chuanxiong reference medicinal material; 3 is ligusticum reference medicinal material; 4-5 are all test samples; 6 is a negative sample lacking chuanxiong; 7 is a negative sample lacking ligusticum; 8 is a double negative sample lacking both chuanxiong and ligusticum.

[0070] Figure 31 Durability test of Ligusticum striatum and Ligusticum chuanxiong (Qingdao silica gel G thin film, T 2.0℃, RH 76%)

[0071] In the figure, 1 is ligustilide reference standard; 2 is chuanxiong reference medicinal material; 3 is ligusticum reference medicinal material; 4-5 are all test samples; 6 is a negative sample lacking chuanxiong; 7 is a negative sample lacking ligusticum; 8 is a double negative sample lacking both chuanxiong and ligusticum.

[0072] Figure 32Thin-layer chromatography (TLC) images of Ligusticum striatum and Ligusticum chuanxiong from 15 batches of samples (1 in the figure is ligustilide reference standard; 2 is Ligusticum striatum reference medicinal material; 3 is Ligusticum chuanxiong reference medicinal material; 4-18 are batch numbers 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, 181204-15, respectively).

[0073] Figure 33 Thin-layer chromatography (TLC) images of Ligusticum striatum and Ligusticum chuanxiong from 15 batches of samples (19 in the figure is ligustilide reference standard; 20 is Ligusticum striatum reference medicinal material; 21 is Ligusticum chuanxiong reference medicinal material; 22-36 are batch numbers 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, 181207-30 respectively).

[0074] Figure 34 Thin-layer chromatogram of roasted licorice root - Method 1 (prepared on Qingdao silica gel G thin-layer plate with 1% sodium hydroxide solution, T22.6℃, RH 12%); 1 in the figure is 2 μl of negative sample lacking roasted licorice root; 2 is 2 μl of test sample; 3

[0075] 4 is 2 μl of lyophilized licorice powder; 5 is 1 μl of glycyrrhizin reference standard; 6 is 5 μl of lyophilized licorice powder.

[0076] 6 is 5 μl of the test sample; 7 is 5 μl of the negative sample of licorice without roasted licorice.

[0077] Figure 35 Thin-layer chromatogram of prepared licorice root - Method 2 (In the figure, 1 is glycyrrhizin reference solution; 2 is prepared licorice root slice solution; 3 is test sample; 4 is negative sample lacking prepared licorice root);

[0078] Figure 36 Thin-layer chromatogram of prepared licorice root - Method 3 (1 in the figure is glycyrrhizin reference standard; 2 is licorice reference material; 3-5 are test samples; 6 is negative sample lacking prepared licorice root);

[0079] Figure 37 Thin-layer chromatography (TLC) of roasted licorice root durability test (Yantai 1% sodium hydroxide silica gel TLC plate, T 14.6℃, RH)

[0080] 49.1%; 1 in the figure is glycyrrhizin reference standard; 2 is licorice reference material; 3-5 are all test samples; 6 is unprocessed.

[0081] Licorice negative sample);

[0082] Figure 38 Thin-layer chromatography (TLC) of roasted licorice root durability test (Qingdao 1% sodium hydroxide silica gel TLC plate, T 14.6℃, RH)

[0083] 49.1%; 1 in the figure is glycyrrhizin reference standard; 2 is licorice reference material; 3-5 are all test samples; 6 is unprocessed.

[0084] Licorice negative sample);

[0085] Figure 39 Thin-layer chromatography (TLC) of licorice root durability test (Qingdao 1% sodium hydroxide silica gel TLC plate, T 15.7℃, RH 75%).

[0086] In the figure, 1 is glycyrrhizin reference standard; 2 is licorice reference material; 3-5 are test samples; 6 is a negative sample of unprocessed licorice.

[0087] Figure 40 Thin-layer chromatography (TLC) of licorice root durability test (Qingdao 1% sodium hydroxide silica gel TLC plate, T 2.0℃, RH 78%).

[0088] In the figure, 1 is glycyrrhizin reference standard; 2 is licorice reference material; 3-5 are test samples; 6 is a negative sample of unprocessed licorice.

[0089] Figure 41 Thin-layer chromatography (TLC) images of processed licorice root from 15 batches of samples (1 in the figure is glycyrrhizin reference standard; 2 is licorice reference material; 3-17 are batch numbers 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, and 181204-15, respectively).

[0090] Figure 42Thin-layer chromatography (TLC) images of processed licorice root from 15 batches of samples (18 in the figure is glycyrrhizin reference standard; 19 is licorice reference material; 20-34 are batch numbers 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, and 181207-30, respectively). Detailed Implementation

[0091] Preparation method of Example 1

[0092] [Prescription] Notopterygium root 4.13g, Ligusticum root 2.06g, Prepared licorice root 2.06g, Angelica pubescens root 4.13g, Saposhnikovia root 2.06g

[0093] Chuanxiong (Ligusticum striatum) 2.06g, Manjingzi (Vitex trifolia) 1.24g

[0094]

Preparation

[0095] Examples 2-4 were used to test the corresponding physical samples prepared in Example 1.

[0096] Example 2 [Identification]

[0097] Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution; separately take 1g of Notopterygium incisum reference material, add 50ml of water and decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; take 1μl each of the test solution and the reference material solution and spot them separately on the same silica gel G thin-layer plate, using a 3:1:2 ratio of n-hexane-60~90℃ petroleum ether-ethyl acetate as the developing solvent, develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference material.

[0098] Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution; take 1g of Angelica pubescens reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; according to the thin-layer chromatography method, take 5μl each of the test solution and the reference material solution, and spot them separately on the same silica gel GF plate.254 On a thin-layer plate, dichloromethane-ethyl acetate in a ratio of 7:4 was used as the developing solvent. The plate was then removed, dried, and sprayed with 10% sulfuric acid ethanol solution. The plate was heated at 105°C until the spots were clearly visible. The plate was then examined under a UV lamp at 254 nm. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position as in the chromatogram of the reference medicinal material.

[0099] Take 2g of the corresponding drug substance, add 20ml of acetone, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.5ml of ethanol to prepare the test solution; separately take 0.2g of Saposhnikovia divaricata reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution in the same way; then take Cimicifuga glycoside reference standard and 5-O-methylvisamidol glycoside reference standard, add ethanol to prepare a mixed solution containing 1mg of each per ml to prepare the reference solution; according to the thin-layer chromatography method, take 6μl of the test solution, 3μl of the reference material solution, and 1μl of the reference solution, and spot them separately on the same silica gel G thin-layer plate, using dichloromethane-methanol in a ratio of 4:1 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, air dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference material and the reference solution.

[0100] Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution; take 0.2g each of Ligusticum striatum and Ligusticum chuanxiong reference materials and prepare Ligusticum striatum and Ligusticum chuanxiong reference material solutions in the same way as the test solution; separately take ligustilide reference standard, add methanol to prepare a solution containing 1mg per 1ml as the reference solution; according to the thin-layer chromatography method, take 5μl each of the test solution, reference material solution and reference solution, and spot them separately on the same silica gel G thin-layer plate, use petroleum ether-ethyl acetate at a ratio of 5:1 at 60-90℃ as the developing solvent, develop, remove, air dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference standard.

[0101] Take 1g of the corresponding drug substance, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution; take 0.2g of licorice reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; separately take glycyrrhizin reference standard, add methanol to prepare a solution containing 1mg per ml to prepare the reference solution; according to the thin-layer chromatography method, take 2μl each of the test solution, reference material solution, and reference solution, and spot them separately on the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution. Use ethyl acetate-formic acid-glacial acetic acid-water in a ratio of 15:1:1:2 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference solution.

[0102] Example 3 [Identification]

[0103] Take 0.5g of the corresponding drug sample, add 25ml of methanol, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1ml of methanol to prepare the test solution; separately take 0.5g of Notopterygium incisum reference material, add 40ml of water and decoct for 25 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; take 1μl each of the test solution and the reference material solution and spot them separately on the same silica gel G thin-layer plate, using a 3:1:2 ratio of n-hexane-60~90℃ petroleum ether-ethyl acetate as the developing solvent, develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference material.

[0104] Take 0.5g of the corresponding drug sample, add 25ml of methanol, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1ml of methanol to prepare the test solution; take 0.5g of Angelica pubescens reference material, add 40ml of water, decoct for 25 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; according to the thin-layer chromatography method, take 5μl each of the test solution and the reference material solution, and spot them separately on the same silica gel GF plate. 254 On a thin-layer plate, dichloromethane-ethyl acetate in a ratio of 7:4 was used as the developing solvent. The plate was then removed, dried, and sprayed with 10% sulfuric acid ethanol solution. The plate was heated at 105°C until the spots were clearly visible. The plate was then examined under a UV lamp at 254 nm. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position as in the chromatogram of the reference medicinal material.

[0105] Take 1g of the corresponding drug substance, add 10ml of acetone, sonicate for 10 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.4ml of ethanol to prepare the test solution; separately take 0.1g of Saposhnikovia divaricata reference material, add 40ml of water, decoct for 25 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution in the same way; then take Cimicifuga glycoside reference standard and 5-O-methylvisamidol glycoside reference standard, add ethanol to prepare a mixed solution containing 1mg of each per ml to prepare the reference solution; according to the thin-layer chromatography method, take 6μl of the test solution, 3μl of the reference material solution, and 1μl of the reference solution, and spot them separately on the same silica gel G thin-layer plate, using dichloromethane-methanol in a ratio of 4:1 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, air dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference material and the reference solution.

[0106] Take 0.5g of the corresponding drug sample, add 25ml of methanol, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1ml of methanol to prepare the test solution; take 0.1g each of Ligusticum striatum and Ligusticum chuanxiong reference materials and prepare Ligusticum striatum and Ligusticum chuanxiong reference material solutions in the same way as the test solution; separately take ligustilide reference standard, add methanol to prepare a solution containing 1mg per 1ml as the reference solution; according to the thin-layer chromatography method, take 5μl each of the test solution, reference material solution and reference solution, and spot them separately on the same silica gel G thin-layer plate, use petroleum ether-ethyl acetate at a ratio of 5:1 at 60-90℃ as the developing solvent, develop, remove, air dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference standard.

[0107] Take 1g of the corresponding drug substance, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution; take 0.2g of licorice reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; separately take glycyrrhizin reference standard, add methanol to prepare a solution containing 1mg per ml to prepare the reference solution; according to the thin-layer chromatography method, take 2μl each of the test solution, reference material solution, and reference solution, and spot them separately on the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution. Use ethyl acetate-formic acid-glacial acetic acid-water in a ratio of 15:1:1:2 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference solution.

[0108] Example 4 [Identification]

[0109] Take 1.5g of the corresponding drug sample, add 35ml of methanol, sonicate for 40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 3ml of methanol to prepare the test solution; separately take 1.5g of Notopterygium incisum reference material, add 60ml of water and decoct for 35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue in the same way; take 1μl of each of the test solution and the reference material solution and spot them separately on the same silica gel G thin-layer plate, develop with a 3:1:2 ratio of n-hexane-60~90℃ petroleum ether-ethyl acetate as the developing solvent, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference material.

[0110] Take 1.5g of the corresponding drug sample, add 35ml of methanol, sonicate for 40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 3ml of methanol to prepare the test solution; take 1.5g of Angelica pubescens reference material, add 60ml of water, decoct for 35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; according to the thin-layer chromatography method, take 5μl each of the test solution and the reference material solution, and spot them separately on the same silica gel GF plate. 254 On a thin-layer plate, dichloromethane-ethyl acetate in a ratio of 7:4 was used as the developing solvent. The plate was then removed, dried, and sprayed with 10% sulfuric acid ethanol solution. The plate was heated at 105°C until the spots were clearly visible. The plate was then examined under a UV lamp at 254 nm. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position as in the chromatogram of the reference medicinal material.

[0111] Take 3g of the corresponding drug substance, add 30ml of acetone, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.6ml of ethanol to prepare the test solution; separately take 0.3g of Saposhnikovia divaricata reference material, add 60ml of water, decoct for 35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution in the same way; then take Cimicifuga glycoside reference standard and 5-O-methylvisamidol glycoside reference standard, add ethanol to prepare a mixed solution containing 1mg of each per ml to prepare the reference solution; according to the thin-layer chromatography method, take 6μl of the test solution, 3μl of the reference material solution, and 1μl of the reference solution, and spot them separately on the same silica gel G thin-layer plate, using dichloromethane-methanol in a ratio of 4:1 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, air dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference material and the reference solution.

[0112] Take 1.5g of the corresponding drug sample, add 35ml of methanol, sonicate for 40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 3ml of methanol to prepare the test solution; take 0.3g each of Ligusticum striatum and Ligusticum chuanxiong reference materials and prepare Ligusticum striatum and Ligusticum chuanxiong reference material solutions in the same way as the test solution; separately take ligustilide reference standard, add methanol to prepare a solution containing 1mg per ml to prepare the reference solution; according to the thin-layer chromatography method, take 5μl each of the test solution, reference material solution and reference solution, and spot them separately on the same silica gel G thin-layer plate, use petroleum ether-ethyl acetate with a ratio of 5:1 at 60-90℃ as the developing solvent, develop, remove, air dry, and examine under ultraviolet light at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference solution.

[0113] Take 1g of the corresponding drug substance, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution; take 0.2g of licorice reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; separately take glycyrrhizin reference standard, add methanol to prepare a solution containing 1mg per ml to prepare the reference solution; according to the thin-layer chromatography method, take 2μl each of the test solution, reference material solution, and reference solution, and spot them separately on the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution. Use ethyl acetate-formic acid-glacial acetic acid-water in a ratio of 15:1:1:2 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference solution.

[0114] To further verify the effectiveness of the present invention, the inventors conducted a series of research experiments, some of which are excerpted below:

[0115] Experimental Example

[0116] 1. Thin-layer chromatography identification study of Qianghuo Shengshi Decoction

[0117] 1.1 [Prescription] Notopterygium root 4.13g, Ligusticum root 2.06g, Prepared licorice root 2.06g, Angelica pubescens root 4.13g, Saposhnikovia root 2.06g

[0118] Chuanxiong (Ligusticum striatum) 2.06g, Manjingzi (Vitex trifolia) 1.24g

[0119] 1.2 [Preparation Method] The above seven ingredients are crushed into coarse granules. According to the dosage for a single dose, weigh each crushed ingredient, place it in a clay pot, add 400ml of water, bring to a boil over high heat, simmer over low heat for 31 minutes, filter, concentrate the filtrate under reduced pressure until the ratio of the amount of medicinal slices to the weight of the concentrate is approximately 1:2 to 1:4 (g:g), freeze dry to obtain a dry powder, package it, and the corresponding physical product is obtained.

[0120] 1.3 Materials

[0121] Imperatorin reference standard: batch number 111821-201604, China National Institutes for Food and Drug Control;

[0122] Notopterygium root slices: Batch number QH-20171228-CAB-YP05;

[0123] Notopterygium incisum reference material: (batch number: 120935-201408, China National Institutes for Food and Drug Control);

[0124] Dihydroapigenin angelic acid ester reference standard: batch number 111583-201605, China National Institutes for Food and Drug Control;

[0125] Angelica pubescens slices: batch number 1609001;

[0126] Osthol reference standard: batch number 110822-201619, China National Institutes for Food and Drug Control;

[0127] Reference material for Angelica pubescens: Batch number 120940-201111, China National Institutes for Food and Drug Control.

[0128] 5-O-methylvisamidol glycoside reference standard: batch number 11523-201610, China National Institutes for Food and Drug Control;

[0129] Saposhnikovia divaricata slices: batch number 1704001;

[0130] Cimicifugoside reference standard: batch number 111522-201712, China National Institutes for Food and Drug Control;

[0131] Reference herb for Saposhnikovia divaricata: Batch No. 120947-201409, China National Institutes for Food and Drug Control.

[0132] Chuanxiong (Ligusticum striatum) slices: Batch number: 1707004;

[0133] Ligusticum striatum slices: Batch number: 1712001;

[0134] Notopterygium root slices: Batch number: 1706002;

[0135] Ferulic acid reference standard, batch number: 110773-201614, China National Institutes for Food and Drug Control;

[0136] Ligustrazine reference standard: batch number 111737-201608, China National Institutes for Food and Drug Control;

[0137] Reference material for Ligusticum chuanxiong: 120918-201612, China National Institutes for Food and Drug Control;

[0138] Ligusticum striatum reference material: batch number 121129-200401, China Institute for Drug and Biological Products.

[0139] Glycyrrhizin reference standard: batch number 111610-201607, China National Institutes for Food and Drug Control;

[0140] Prepared licorice root freeze-dried powder: batch number 2018022603;

[0141] Licorice reference material: batch number 120904-201620, China National Institutes for Food and Drug Control;

[0142] Silicone G Thin Film: Yantai Jiangyou Silicone Development Co., Ltd. and Qingdao Ocean Chemical Plant;

[0143] Silicone G thin film: Qingdao Ocean Chemical Plant.

[0144] 2. Study on thin-layer chromatography method of Notopterygium incisum

[0145] 2.1 Establishment of Thin-Layer Chromatography Method

[0146] Method 1 refers to the method under the "Identification" section of Part I of the 2015 edition of the Chinese Pharmacopoeia:

[0147] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 5ml of methanol to prepare the test solution. Separately, take 18 negative samples lacking Notopterygium incisum and prepare a negative sample solution using the same method. Separately, take 1g of Notopterygium incisum powder, place it in a round-bottom flask, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare a Notopterygium incisum solution using the same method starting from "adding 30ml of methanol". Also, take the reference standard of purslane, add methanol to prepare a solution containing 0.5mg per ml, and use this as the reference solution. Perform thin-layer chromatography, taking 3μl of each of the above four solutions and spotting them separately on the same silica gel G thin-layer plate. Use chloroform-methanol (8:2) as the developing solvent, develop, remove, air dry, and examine under ultraviolet light (365nm). The results are shown in the figure. Figure 1 .

[0148] Results Analysis: Figure 1 It can be seen that in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the Notopterygium root slices and the reference sample. However, the negative sample lacking Notopterygium root shows interference at the corresponding positions, which requires further study.

[0149] Method 2 considers that in addition to ultraviolet light (365nm), the inspection conditions often involve the use of a certain concentration of vanillin-sulfuric acid solution for color development. Therefore, the color development conditions were changed and the system was studied, as follows:

[0150] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution. Separately, take 1g of a negative sample lacking Notopterygium incisum and prepare a negative sample solution using the same method. Take 1g of Notopterygium incisum reference material, add 50ml of water and decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution using the same method starting from "adding 30ml of methanol". Apply 1μl of each of the above solutions to the same silica gel G thin-layer plate using thin-layer chromatography. Develop with n-hexane-petroleum ether (60-90℃)-ethyl acetate (3:1:2) as the developing solvent. Remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. The results are shown in the figure. Figure 2 .

[0151] Results Analysis: Figure 2 It can be seen that in the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference medicinal material, and there is no negative interference. Therefore, the method is feasible, and the sample volume is determined to be 1 μl each for the test sample solution and the reference medicinal material solution.

[0152] 2.2 Durability Test

[0153] Durability tests were conducted according to "Method 2" under section "2.1", examining the effects of different manufacturers' silicone G thin-layer plates (Yantai Jiangyou Silicone Development Co., Ltd. and Qingdao Ocean Chemical Plant), as well as different development temperatures and relative humidity. The results showed good durability under all development conditions, with clear main spots. See the results below. Figure 3 , Figure 4 , Figure 5 , Figure 6 Therefore, this thin-layer method is included in the main text.

[0154] 2.3 Method Determination

[0155] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution. Separately, take 1g of Notopterygium incisum reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the step of "adding 30ml of methanol". Apply 1μl of each of the above solutions to the same silica gel G thin-layer plate using thin-layer chromatography. Develop with hexane-petroleum ether (60-90℃)-ethyl acetate (3:1:2) as the developing solvent. Remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color as the reference material spot should appear at the corresponding position.

[0156] 2.4 Thin-layer chromatography identification of multiple batches of corresponding physical samples

[0157] Take the reference herb solution of Notopterygium incisum (numbered 1 and 17) and the test solution (numbered 2-16) respectively. The corresponding sample batch numbers are: 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, 181204-1 5; Sample batch numbers corresponding to numbers 18-32 are as follows: 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, 181207-30). Identification was performed according to the thin-layer chromatography identification method under section “2.3”. The results are shown in […]. Figure 7 and Figure 8 .

[0158] Experimental results: Figure 7 and Figure 8 It can be seen that in the chromatogram of the test sample, the main spot of the same color is displayed at the corresponding position as in the chromatogram of the reference medicinal material.

[0159] 3. Study on thin-layer chromatography method of Angelica pubescens

[0160] 3.1 Establishment of Thin-Layer Chromatography Method

[0161] Method 1 refers to the method under the "Identification" section of Part I of the 2015 edition of the Chinese Pharmacopoeia:

[0162] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 5ml of methanol to prepare the test solution. Separately, take 1g of a negative sample lacking Angelica pubescens and prepare a negative sample solution using the same method. Separately, take 1g of Angelica pubescens slices, place them in a round-bottom flask, add 50ml of water, reflux for 30 minutes, centrifuge, collect the supernatant, evaporate to dryness, and prepare a solution of Angelica pubescens slices using the same method, starting from "add 30ml of methanol". Also, take osthol and dihydroaperitol angelica ester reference standards, and prepare reference solutions containing 0.5mg per ml using methanol. According to the thin-layer chromatography method, take 2 μl each of the test solution and the negative solution lacking Angelica pubescens, 5 ml of Angelica pubescens slice solution, and 1 μl each of the reference solution, and spot them separately on the same silica gel G thin-layer plate. Use petroleum ether (60-90℃)-ethyl acetate (7:3) as the developing solvent, develop, remove, air dry, and examine under ultraviolet light (365nm). The results are shown in the figure. Figure 9 .

[0163] Results Analysis: Figure 9 It can be seen that in the chromatogram of the test sample, the fluorescent spot at the corresponding position of the osthol reference standard is lighter; at the corresponding position of the dihydroaperitol angelic acid ester reference standard, the fluorescent spot of the same color appears, but there is interference in the negative, so this method is not feasible.

[0164] Method 2: Based on the technical issues identified in the "Method", relevant literature was reviewed and analyzed. The developing solvent system for *Notopterygium incisum* was replaced with dichloromethane-ethyl acetate (7:4). A 10% sulfuric acid ethanol solution was sprayed on top, and the mixture was heated at 105°C until the spots were clearly visible. The spots were then examined under ultraviolet light (365 nm). Results are shown below. Figure 10 .

[0165] Results Analysis: Figure 10 It can be seen that the chromatogram of the test sample shows fluorescent spots of the same color at the corresponding positions as the chromatogram of Angelica pubescens slices, but the negative result is an interference, making the method unfeasible.

[0166] Method 3 analyzes the technical problems existing in "Method 2" and changes the inspection conditions: Ultraviolet light (254nm) is used for testing. Take 1g of the sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution. Take another 1g of a negative sample lacking Angelica pubescens and prepare a negative sample solution using the same method. Take another 1g of Angelica pubescens reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare a reference material solution using the same method starting from "adding 30ml of methanol". According to thin-layer chromatography, take 5μl of each of the above three solutions and spot them separately on the same silica gel GF plate. 254 On thin-layer plates, dichloromethane-ethyl acetate (7:4) was used as the developing solvent. The plates were then removed, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105℃ until the spots were clearly visible. The plates were then examined under ultraviolet light (254 nm). Results are shown below. Figure 11 .

[0167] Experimental results: Figure 11 It can be seen that in the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference medicinal material, and there is no negative interference. Therefore, the method is feasible, and the sample loading is determined to be 5 μl each for the reference medicinal material solution and the test sample solution.

[0168] 3.2 Durability Assessment

[0169] Durability tests were conducted according to the methods described above, examining thin-layer plates from different manufacturers (Yantai Jiangyou Silicone Development Co., Ltd. and Qingdao Ocean Chemical Plant), as well as different development temperatures and relative humidity levels. The results showed good durability under all development conditions, with clear main spots. (See attached figures.) Figure 12 , Figure 13 , Figure 14 , Figure 15Therefore, this thin-layer method is included in the main text.

[0170] 3.3 Method Determination

[0171] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, and add 2ml of methanol to the residue.

[0172] Dissolve the sample to prepare the test solution. Separately, take 1g of Angelica pubescens reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue (starting with "add 30ml of methanol"). Apply 5μl of each of the two solutions to the same silica gel GF plate using thin-layer chromatography. 254 On a thin-layer plate, dichloromethane-ethyl acetate (7:4) was used as the developing solvent. The plate was then removed, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105℃ until the spots were clearly visible. The plate was then examined under ultraviolet light (254 nm). The test sample chromatogram showed a main spot of the same color at the corresponding position as the reference medicinal material chromatogram.

[0173] 3.4 Thin-layer chromatography identification of multiple batches of corresponding physical samples

[0174] Take the reference herb solution of Angelica pubescens (numbered 1 and 17) and the test solution (numbered 2-16), with the corresponding sample batch numbers as follows: 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, 181204-1 5; Sample batch numbers corresponding to numbers 18-32 are as follows: 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, 181207-30). Identification was performed according to the thin-layer chromatography identification method under section “3.3”. The results are shown in […]. Figure 16 and Figure 17 .

[0175] Experimental results: Figure 16 and Figure 17 It can be seen that in the chromatogram of the test sample, the main spot of the same color is displayed at the corresponding position as in the chromatogram of the reference medicinal material.

[0176] 4. Study on thin-layer chromatography method for Saposhnikovia divaricata

[0177] 4.1 Establishment of Thin-Layer Chromatography Method

[0178] Method 1: Refer to the method under "Identification" of Saposhnikovia divaricata in Part I of the 2015 edition of the Chinese Pharmacopoeia: Take 2g of this product, add 20ml of acetone, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1ml of ethanol to prepare the test solution. Separately, take 2g of a negative sample lacking Saposhnikovia divaricata and prepare a negative sample solution using the same method. Then, take 0.3g of Saposhnikovia divaricata slices, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 20ml of acetone to prepare a Saposhnikovia divaricata slice solution using the same method. Next, take cimicifugoside reference standard and 5-O-methylvisamidolol reference standard, and prepare reference solutions containing 1mg each per 1ml using ethanol. For thin-layer chromatography, take 7μl each of the test solution, the negative sample solution lacking Saposhnikovia divaricata, 5μl of the Saposhnikovia divaricata slice solution, and 2μl of the reference solution, and spot them separately on the same silica gel GF plate. 254 On the thin-layer plate, chloroform-methanol (4:1) was used as the developing solvent. The plate was then removed, dried, and examined under ultraviolet light (254 nm). The results are shown below. Figure 18 .

[0179] Results Analysis: Figure 18 It can be seen that the chromatographic band of the test sample is relatively dark, and the chromatogram of the test sample is almost identical to that of the negative chromatogram, indicating that there is interference from the negative chromatogram. The method needs to be further optimized.

[0180] Method 2 addresses the technical issues of Method 1 and analyzes relevant literature, focusing on the developing solvent system and colorimetric conditions, as detailed below:

[0181] Take 2g of this product, add 20ml of acetone, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.5ml of ethanol to prepare the test solution. Separately, take 2g of a negative sample lacking *Saposhnikovia divaricata*, and prepare a negative sample solution using the same method. Next, take 0.2g of *Saposhnikovia divaricata* reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare a reference material solution using the same method starting from "adding 20ml of acetone". Then, take *Cimicifuga foetida* glycoside reference standard and 5-O-methylvisamidolol glycoside reference standard, and prepare reference solution containing 1mg per ml of each with ethanol. For thin-layer chromatography, apply 5 μl, 6 μl, and 7 μl of the test solution, 7 μl of the negative sample solution for *Saposhnikovia divaricata* (if missing), and 1 μl, 2 μl, and 3 μl of the reference herb solution and reference solution, respectively, onto the same silica gel G thin-layer plate. Develop the plate using dichloromethane-methanol (4:1) as the developing solvent. Remove the plate, air-dry it, spray it with 10% sulfuric acid in ethanol solution, air-dry it again, and examine it under ultraviolet light (365 nm). The results are shown in the figure. Figure 19 .

[0182] Results Analysis: Figure 19It can be seen that in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference medicinal material and the reference standard, and there is no negative interference. Therefore, the method is feasible, and the sample loading is determined to be: 1 μl of the reference standard solution, 3 μl of the reference medicinal material solution, and 6 μl of the test sample solution.

[0183] 4.2 Durability Test

[0184] Durability tests were conducted according to the methods described above, examining thin-layer plates from different manufacturers (Yantai Jiangyou Silicone Development Co., Ltd. and Qingdao Ocean Chemical Plant), as well as the effects of different development temperatures and relative humidity. The results showed good durability under all development conditions, with clear main spots. Figure 20 , Figure 21 , Figure 22 , Figure 23 Therefore, this thin-layer method is included in the main text.

[0185] 4.3 Method Determination

[0186] Take 2g of this product, add 20ml of acetone, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.5ml of ethanol to prepare the test solution. Separately, take 0.2g of Saposhnikovia divaricata reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution using the same method starting from "adding 20ml of acetone". Then, take cimicifugoside reference standard and 5-O-methylvisamidolol reference standard, and prepare solutions containing 1mg each per ml with ethanol to prepare reference solutions. Apply 6μl of the test solution, 3μl of the reference material solution, and 1μl of the reference solution to the same silica gel G thin-layer plate using thin-layer chromatography. Develop with dichloromethane-methanol (4:1) as the developing solvent, remove, air dry, spray with 10% sulfuric acid ethanol solution, air dry, and examine under ultraviolet light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference medicinal material and the reference standard.

[0187] 4.4 Identification of multiple batches of thin-layer chromatography

[0188] Take cimicifugoside reference standards (numbered 1 and 19), 5-O-methylvisamidolol reference standards (numbered 2 and 20), and Saposhnikovia divaricata reference materials (numbered 3 and 21) and test solutions (numbered 4-18). The corresponding sample batch numbers are: 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, and 181204-1. 3. Sample batch numbers 181204-14 and 181204-15; the corresponding sample batch numbers for batches 22-36 are: 181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, and 181207-30. Identification was performed according to the thin-layer chromatography method under section “4.3”. The results are shown in […]. Figure 24 and Figure 25 .

[0189] Experimental results: Figure 24 and Figure 25 It can be seen that in the chromatogram of the test sample, fluorescent spots of the same color are displayed at the corresponding positions as those of the reference standard and the reference medicinal material.

[0190] 5. Study on thin-layer chromatography methods for Ligusticum striatum and Ligusticum chuanxiong

[0191] 5.1 Establishment of Thin-Layer Chromatography Method

[0192] Since both Ligusticum striatum and Ligusticum chuanxiong contain indicative components such as ferulic acid and ligustilide, the two herbs were combined for thin-layer chromatography identification.

[0193] Method 1: The test sample is ultrasonically analyzed using methanol. The developing system is adjusted as follows:

[0194] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, cool, filter, evaporate the filtrate to dryness, dissolve the residue in 5ml of methanol to prepare the test solution. Separately, take 18 negative samples lacking Ligusticum chuanxiong, 18 negative samples lacking Ligusticum sinense, and 18 negative samples lacking both Ligusticum chuanxiong and Ligusticum sinense, and prepare their respective negative sample solutions using the same method. Separately, take 1g of Notopterygium incisum slices, 0.5g of Ligusticum chuanxiong slices, and 0.5g of Ligusticum sinense slices, place them in round-bottom flasks, add 50ml of water, reflux for 30 minutes, centrifuge, collect the supernatant, evaporate to dryness, and prepare the Notopterygium incisum, Ligusticum chuanxiong, and Ligusticum sinense slice solutions using the same method (add 30ml of methanol). Take ferulic acid reference standard, add methanol to prepare a solution containing 1mg per ml, as the ferulic acid reference standard solution. Take ligustilide reference standard, add methanol to prepare a solution containing 0.1mg per ml, as the ligustilide reference standard solution. Apply 2 μl each of the test solution, negative solutions, and reference solutions, and 5 μl of the herbal medicine solution to the same silica gel G thin-layer plate using the thin-layer chromatography method. Develop the plate using toluene-chloroform-ethyl acetate-glacial acetic acid (6:5:2:1) as the developing solvent. Remove the plate, air-dry it, and examine it under ultraviolet light (365 nm). Results are shown below. Figure 26 .

[0195] Results Analysis: Figure 26 It is known that ferulic acid is present in the processed slices of Ligusticum chuanxiong, Ligusticum striatum, and Notopterygium incisum. This component lacks specificity and is of little qualitative significance, therefore it is not included in the standard. However, the chromatograms of the test samples show fluorescent spots of the same color at the corresponding positions of ligustilide, and the absence of negative samples from Ligusticum striatum and Ligusticum chuanxiong indicates no interference, suggesting the method is basically feasible. However, the chromatograms of Ligusticum striatum and Ligusticum chuanxiong slices show no spots at the corresponding positions, possibly due to the decoction process. Further research will be conducted without decoction of the slices. Furthermore, the R-value for ligustilide is too high under this developing system; a more systematic approach will be used in the next step.

[0196] Method 2 addresses the technical issues of Method 1 by adjusting the preparation method of the reference medicinal material and the developing solvent ratio system, as follows:

[0197] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution. Separately, take 1g each of negative samples lacking Ligusticum striatum, negative samples lacking Ligusticum chuanxiong, and negative samples lacking both Ligusticum striatum and Ligusticum chuanxiong, and prepare the corresponding negative sample solutions using the same method. Take 0.2g each of Ligusticum striatum (Liaoning Ligusticum striatum) and Ligusticum chuanxiong reference materials, and prepare the reference material solution using the same method. Take ligustilide reference standard, add methanol to prepare a solution containing 1mg per ml, and use this as the reference solution. Apply 2μl, 3μl, and 5μl of each test solution, reference material solution, and reference solution, and 5μl of each negative sample solution to the same silica gel G thin-layer plate using thin-layer chromatography. Develop with petroleum ether (60-90℃)-ethyl acetate (5:1), remove, air dry, and examine under ultraviolet light (365nm). Results are shown below. Figure 27 .

[0198] Results Analysis: Figure 27 It can be seen that the test sample chromatogram shows fluorescent spots of the same color at the corresponding positions as the reference medicinal material chromatogram and the reference standard, and there is no negative interference. Therefore, the method is feasible, and the sample loading is determined to be 5 μl each of the reference standard solution, the reference medicinal material solution and the test sample solution.

[0199] 5.2 Durability Test

[0200] Durability tests were conducted according to the methods described above, examining thin-layer plates from different manufacturers (Yantai Jiangyou Silicone Development Co., Ltd. and Qingdao Ocean Chemical Plant), as well as different development temperatures and relative humidity levels. The results showed good durability under all development conditions, with clear main spots. (See attached figures.) Figure 28 , Figure 29 , Figure 30 , Figure 31 Therefore, this thin-layer method is included in the main text.

[0201] 5.3 Method Determination

[0202] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution. Separately, take 0.2g each of Ligusticum striatum (Liaoning Ligusticum striatum) and Ligusticum chuanxiong reference materials, and prepare reference material solutions using the same method. Also, take ligustilide reference standard, add methanol to prepare a solution containing 1mg per ml, and prepare the reference standard solution. Apply 5μl of the above four solutions to the same silica gel G thin-layer plate using thin-layer chromatography. Develop with petroleum ether (60-90℃)-ethyl acetate (5:1) as the developing solvent, remove, air dry, and examine under ultraviolet light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference materials and the reference standard.

[0203] 5.4 Thin-layer chromatography identification of multiple batches of material references

[0204] Take ligustilide reference standards (numbered 1 and 19), 5-O-methylvisamidol glycoside reference standards (numbered 2 and 20), and Saposhnikovia divaricata reference medicinal materials (numbered 3 and 21) and test solutions (numbered 4-18). The corresponding sample batch numbers are: 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, and 181204-15, respectively. Sample batch numbers 22-36 correspond to the following batch numbers in sequence: 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, and 181207-30. They were identified according to the thin-layer chromatography identification method under section “5.3”, and the results are shown below. Figure 32 and Figure 33 .

[0205] Experimental results: 32 and Figure 33 It can be seen that in the chromatogram of the test sample, fluorescent spots of the same color are displayed at the corresponding positions as those of the reference standard and the reference medicinal material.

[0206] 6. Study on thin-layer chromatography method of roasted licorice

[0207] 6.1 Establishment of Thin-Layer Chromatography Method

[0208] Method 1 follows the method described in the "Identification" section of the 2015 edition of the Chinese Pharmacopoeia, Part I: Take 1g of the sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 40ml of water, extract three times with 20ml of n-butanol each time, combine the n-butanol extracts, wash three times with water, discard the aqueous solution, evaporate the n-butanol extract to dryness, dissolve the residue in 5ml of methanol, and use this as the test solution. Separately, take 1g of a negative sample of unprocessed licorice and prepare a negative sample solution using the same method. Then, take 0.25g of lyophilized licorice powder and prepare a lyophilized licorice powder solution using the same method. Finally, take glycyrrhizin reference standard and prepare a solution containing 2mg per ml using methanol, as the reference solution. Apply 2 μl and 5 μl of the test solution, 2 μl and 5 μl of the negative solution for licorice root deficiency, 2 μl and 5 μl of the lyophilized licorice powder solution, and 1 μl of the reference solution to the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution. Develop the plate using ethyl acetate-formic acid-glacial acetic acid-water (15:1:1:2) as the developing solvent. Remove the plate, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible. Examine the plate under sunlight and ultraviolet light (365 nm). The results are shown in the figure. Figure 34 .

[0209] Results Analysis: Figure 34 It can be seen that, under sunlight, the chromatogram of the test sample shows spots of the same color at the corresponding positions as the chromatograms of the reference sample and the prepared licorice root slices, and there is no negative interference, but the spots in the chromatogram of the test sample are lighter in color; under ultraviolet light (365nm), the chromatogram of the test sample does not show fluorescent spots of the same color as the chromatograms of the reference sample and the prepared licorice root slices, so the method is not feasible, and the preparation method of the test sample should be changed.

[0210] Method 2, based on Method 1, involves purifying the test sample, as detailed below:

[0211] Take 2g of this product, dissolve it in 40ml of water, and extract it three times with 20ml of water-saturated n-butanol each time. Combine the n-butanol extracts, wash twice with 30ml of ammonia solution each time, combine the ammonia solution, adjust the pH to 3-4 with hydrochloric acid, and then extract it twice with 30ml of ethyl acetate each time. Combine the ethyl acetate extracts, evaporate to dryness, and dissolve the residue in 1ml of methanol to obtain the test solution. Separately, take 2g of a negative sample lacking prepared licorice root and prepare a negative sample solution for prepared licorice root deficiency using the same method. Separately, take 1g of prepared licorice root slices, add 50ml of water, decoct for 30 minutes, filter, and prepare a prepared licorice root slice solution from the step of "extracting three times with water-saturated n-butanol" using the same method. Also, take glycyrrhizin reference standard, add methanol to prepare a solution containing 2mg per ml, and use this as the reference solution. Apply 1 μl of each of the four solutions described above to the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution according to the thin-layer chromatography method. Develop the plate using ethyl acetate-formic acid-glacial acetic acid-water (15:1:1:2) as the developing solvent. Remove the plate, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible. Examine the plate under ultraviolet light (365 nm). The results are shown in the figure. Figure 35 .

[0212] Results Analysis: Figure 35 It is known that under sunlight and ultraviolet light (365nm), the chromatogram of the test sample shows spots or fluorescent spots of the same color at the corresponding positions as those of the prepared licorice root slices and the reference solution, indicating no negative interference and the method is feasible. However, due to the complexity of the test sample preparation method, a simplified preparation method is proposed.

[0213] Method 3 simplifies the sample preparation method to direct ultrasonication with methanol, as follows:

[0214] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution. Separately, take 18 negative samples of unprocessed licorice and prepare a negative sample solution of unprocessed licorice using the same method. Take 0.2g of licorice (licorice) reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution using the same method starting from "add 30ml of methanol". Separately, take glycyrrhizin reference standard and prepare a solution containing 1mg per ml using methanol to prepare the reference solution. Apply 2μl of each of the above solutions to the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution according to the thin-layer chromatography method. Use ethyl acetate-formic acid-glacial acetic acid-water (15:1:1:2) as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible. The results are shown in the figure. Figure 36 .

[0215] Results Analysis: Figure 36 It can be seen that under sunlight, the chromatogram of the test sample shows four identical yellow spots at the corresponding positions as the chromatogram of the reference medicinal material, and there is no interference. Under ultraviolet light (365nm), the chromatogram of the test sample mainly shows three identical yellow-green fluorescent spots at the corresponding positions as the chromatogram of the reference medicinal material, which are negative and have no interference. The fluorescent spots of other colors are negative and have interference. That is, the effective chromatographic information of ultraviolet light (365nm) is less than that of sunlight, and the chromatographic information of the two inspection conditions is not complementary. Therefore, the inspection condition is determined to be sunlight, and the sample volume is determined to be 2μl each of the reference standard, the reference medicinal material solution, and the test sample solution.

[0216] 6.2 Durability Test

[0217] Durability tests were conducted according to the methods described above, examining thin-layer plates from different manufacturers (Yantai Jiangyou Silicone Development Co., Ltd. and Qingdao Ocean Chemical Plant), as well as the effects of different development temperatures and relative humidity. The results showed good durability under all development conditions, with clear spots. (See attached image.) Figure 37 , Figure 38 , Figure 39 , Figure 40 Therefore, this thin-layer method is included in the main text.

[0218] 6.3 Method Determination

[0219] Take 1g of this product, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution. Separately, take 0.2g of licorice (Glycyrrhiza) reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the "add 30ml of methanol" step onwards using the same method. Also, take glycyrrhizin reference standard, add methanol to prepare a solution containing 1mg per ml, and use this as the reference solution. Apply 2μl of each of the above three solutions to the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution using thin-layer chromatography. Develop with ethyl acetate-formic acid-glacial acetic acid-water (15:1:1:2) as the developing solvent. Remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference standard.

[0220] 6.4 Thin-layer chromatography identification of multiple batches of material references

[0221] Glycyrrhizin reference standards (numbered 1 and 18), licorice reference medicinal materials (numbered 2 and 19), and test solutions (numbered 3-17) were taken respectively. The corresponding sample batch numbers are: 181130-01, 181130-02, 181130-03, 181130-04, 181130-05, 181130-06, 181130-07, 181130-08, 181203-09, 181130-10, 181203-11, 181203-12, 181204-13, 181204-14, 1 Samples 81204-15; the corresponding batch numbers for samples 20-34 are: 1181204-16, 181204-17, 181204-18, 181205-19, 181205-20, 181205-21, 181205-22, 181206-23, 181206-24, 181206-25, 181206-26, 181207-27, 181207-28, 181207-29, 181207-30. They were identified according to the thin-layer chromatography identification method under section “6.3”, and the results are shown below. Figure 41 and Figure 42 .

[0222] Experimental results: consisting of 41 and Figure 42 It can be seen that in the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference medicinal material and the reference standard.

[0223] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A method for detecting a wind-dispelling and dampness-removing drug, wherein the prescription and preparation method of the drug are as follows: Prescription: Notopterygium root 4.13g, Ligusticum root 2.06g, Prepared licorice root 2.06g, Angelica pubescens root 4.13g, Saposhnikovia root 2.06g Chuanxiong (Ligusticum striatum) 2.06g, Manjingzi (Vitex trifolia) 1.24g Preparation: Crush the above seven ingredients into coarse particles. Weigh each crushed ingredient according to the single-dose dosage, place them in a clay pot, add 400ml of water, bring to a boil over high heat, simmer over low heat for 31 minutes, filter, concentrate the filtrate under reduced pressure until the ratio of the amount of medicinal slices to the weight of the concentrated liquid is 1:2 to 1:4, freeze dry to obtain dry powder, package, and obtain the corresponding physical product. Its features are, The detection methods for the drugs are as follows: thin-layer chromatography identification of Notopterygium incisum; thin-layer chromatography identification of Angelica pubescens; thin-layer chromatography identification of Saposhnikovia divaricata; thin-layer chromatography identification of Ligusticum striatum and Ligusticum chuanxiong; and thin-layer chromatography identification of Glycyrrhiza uralensis (processed).

2. The detection method according to claim 1, characterized in that, The thin-layer chromatography method for identifying Notopterygium incisum is as follows: Take 0.5-1.5g of the corresponding drug sample, add 25-35ml of methanol, sonicate for 20-40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-3ml of methanol to prepare the test solution; separately take 0.5-1.5g of Notopterygium incisum reference material, add 40-60ml of water and decoct for 25-35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; take 1μl each of the test solution and the reference material solution and spot them separately on the same silica gel G thin-layer plate, using a 3:1:2 ratio of n-hexane-60-90℃ petroleum ether-ethyl acetate as the developing solvent, develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference material.

3. The detection method according to claim 2, characterized in that, The thin-layer chromatography method for identifying Notopterygium incisum is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution; Take 1g of Notopterygium incisum reference material, add 50ml of water and decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Take 1μl each of the test solution and the reference material solution and spot them separately on the same silica gel G thin-layer plate, using a 3:1:2 ratio of n-hexane-60~90℃ petroleum ether-ethyl acetate as the developing solvent, develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position as in the chromatogram of the reference material.

4. The detection method according to claim 1, characterized in that, The thin-layer chromatography method for identifying Angelica pubescens is as follows: Take 0.5-1.5g of the corresponding drug sample, add 25-35ml of methanol, sonicate for 20-40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-3ml of methanol to prepare the test solution; Take 0.5-1.5g of Angelica pubescens reference material, add 40-60ml of water, decoct for 25-35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Apply 5μl each of the test solution and the reference material solution to the same silica gel GF plate according to the thin-layer chromatography method. 254 On a thin-layer plate, dichloromethane-ethyl acetate in a ratio of 7:4 was used as the developing solvent. The plate was then removed, dried, and sprayed with 10% sulfuric acid ethanol solution. The plate was heated at 105°C until the spots were clearly visible. The plate was then examined under a UV lamp at 254 nm. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position as in the chromatogram of the reference medicinal material.

5. The detection method according to claim 4, characterized in that, The thin-layer chromatography method for identifying Angelica pubescens is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to prepare the test solution; Take 1g of Angelica pubescens reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Apply 5μl each of the test solution and the reference material solution to the same silica gel GF plate according to the thin-layer chromatography method. 254 On a thin-layer plate, dichloromethane-ethyl acetate in a ratio of 7:4 was used as the developing solvent. The plate was then removed, dried, and sprayed with 10% sulfuric acid ethanol solution. The plate was heated at 105°C until the spots were clearly visible. The plate was then examined under a UV lamp at 254 nm. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position as in the chromatogram of the reference medicinal material.

6. The detection method according to claim 1, characterized in that, The thin-layer chromatography method for identifying Saposhnikovia divaricata is as follows: Take 1-3g of the corresponding drug sample, add 10-30ml of acetone, sonicate for 10-30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.4-0.6ml of ethanol to prepare the test solution; Separately, take 0.1-0.3g of Saposhnikovia divaricata reference material, add 40-60ml of water, decoct for 25-35 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Then, take cimicifugoside reference standard and 5-O-methylvisamidolol reference standard, add ethanol to prepare... Prepare a mixed solution containing 1 mg of each of the test sample and the reference material solution per 1 ml as the reference solution. Using thin-layer chromatography, apply 6 μl of the test sample solution, 3 μl of the reference material solution, and 1 μl of the reference solution separately to the same silica gel G thin-layer plate. Develop the plate using a 4:1 dichloromethane-methanol mixture as the developing solvent. Remove the plate, air-dry it, spray it with 10% sulfuric acid ethanol solution, air-dry it again, and examine it under a 365 nm UV lamp. In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference solution.

7. The detection method according to claim 1, characterized in that, The thin-layer chromatography method for identifying Saposhnikovia divaricata is as follows: Take 2g of the corresponding drug sample, add 20ml of acetone, sonicate for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 0.5ml of ethanol to obtain the test solution; separately, take 0.2g of Saposhnikovia divaricata reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; then, take cimicifugoside reference standard and 5-O-methylvisamidolol reference standard, and prepare a solution containing 1mg of each per 1ml with ethanol. The mixed solution was used as the reference solution. Using thin-layer chromatography, 6 μl of the test solution, 3 μl of the reference medicinal material solution, and 1 μl of the reference solution were spotted separately onto the same silica gel G thin-layer plate. The plate was developed using a 4:1 dichloromethane-methanol mixture as the developing solvent. The plate was then removed, air-dried, sprayed with 10% sulfuric acid ethanol solution, air-dried again, and examined under a 365 nm UV lamp. Fluorescent spots of the same color appeared at the corresponding positions in the chromatograms of the test sample, the reference medicinal material, and the reference solution.

8. The detection method according to claim 5, characterized in that, The thin-layer chromatography method for identifying Ligusticum striatum and Ligusticum chuanxiong is as follows: Take 0.5-1.5g of the corresponding drug sample, add 25-35ml of methanol, sonicate for 20-40 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-3ml of methanol to prepare the test solution; Take 0.1-0.3g each of Ligusticum striatum and Ligusticum chuanxiong reference materials to prepare the Ligusticum striatum and Ligusticum chuanxiong reference material solutions in the same way as the test solution; Separately take ligustilide reference standard, add methanol to prepare a solution containing 1mg per ml to prepare the reference solution; Take 5μl each of the test solution, reference material solution, and reference solution according to the thin-layer chromatography method, spot them separately on the same silica gel G thin-layer plate, use petroleum ether-ethyl acetate at a ratio of 5:1 at 60-90℃ as the developing solvent, develop, remove, air dry, and examine under a UV lamp at 365nm; In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference materials and the reference solution.

9. The detection method according to claim 8, characterized in that, The thin-layer chromatography method for identifying Ligusticum striatum and Ligusticum chuanxiong is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution; take 0.2g each of Ligusticum striatum and Ligusticum chuanxiong reference materials and prepare the Ligusticum striatum and Ligusticum chuanxiong reference material solutions in the same way as the test solution; separately take ligustilide reference standard, add methanol to prepare a solution containing 1mg per 1ml to obtain the reference solution; according to the thin-layer chromatography method, take 5μl each of the test solution, reference material solution, and reference solution, and spot them separately on the same silica gel G thin-layer plate, use petroleum ether-ethyl acetate at a ratio of 5:1 at 60-90℃ as the developing solvent, develop, remove, air dry, and examine under a UV lamp at 365nm; in the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference materials and the reference solution.

10. The detection method according to claim 1, characterized in that, The thin-layer chromatography method for identifying prepared licorice is as follows: Take 1g of the corresponding drug sample, add 30ml of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol to obtain the test solution; Take 0.2g of licorice reference material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, and prepare the reference material solution from the residue using the same method; Separately, take glycyrrhizin reference standard, add methanol to prepare a solution containing 1mg per ml to obtain the reference standard solution; According to the thin-layer chromatography method, take 2μl each of the test solution, reference material solution, and reference standard solution, and spot them separately on the same silica gel G thin-layer plate prepared with 1% sodium hydroxide solution. Use ethyl acetate-formic acid-glacial acetic acid-water in a ratio of 15:1:1:2 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible; In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference standard.

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