Thin-layer identification method for index components in nerve-calming and mind-tranquilizing pills

Abstract

The invention provides a thin-layer identification method for index components in nerve-calming and mind-tranquilizing pills, and relates to the technical field of pharmaceutical raw material detection and traditional Chinese medicine quality control. The thin-layer identification method comprises the following steps: S1, sample application: sucking a test solution of the nerve soothing and mind tranquilizing pills, a negative test solution of the nerve soothing and mind tranquilizing pills, a reference medicinal material solution and a reference substance solution, and applying the test solution, the reference medicinal material solution and the reference substance solution on a silica gel G thin-layer plate; s2, spreading: spreading the silica gel G thin-layer plate by using a spreading agent, taking out the silica gel G thin-layer plate after spreading, and airing the silica gel G thin-layer plate; and S3, inspecting: spraying a developing solution on the aired silica gel G thin-layer plate for developing inspection. According to the method disclosed by the invention, eight index components including ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rf, tenuifolin, dehydrotumulin, pachymic acid and beta-asarone in the nerve calming and mind stabilizing pill can be identified only by preparing three test sample solutions, and the method is good in applicability, clear in chromatogram and good in separation degree.

Description

Technical Field

[0001] The invention belongs to the technical fields of pharmaceutical raw material detection technology and traditional Chinese medicine quality control technology, and in particular relates to a thin layer identification method for index components in Anshen Dingzhi Pills. Background Art

[0002] Anshen Dingzhi Pills (prescription) comes from Cheng Guopeng's "Medical Enlightenment" (Volume 4. Not allowed to lie down) in the Qing Dynasty. It is composed of Poria cocos, Poria cocos, ginseng, Polygala tenuifolia, Acorus calamus, and dragon's tooth. It is made into pills with refined honey and coated with cinnabar. Anshen Dingzhi Pills have the effects of calming the mind and stabilizing the spirit, replenishing qi and calming the nerves. It is a classic prescription for doctors of all dynasties to treat diseases caused by anxiety, depression, panic, etc. caused by mental malnutrition.

[0003] Thin layer chromatography is an important experimental technique for rapid separation and qualitative analysis of small amounts of substances. The equipment and operation process are relatively simple, and the sample dosage and time are relatively low. However, the existing thin layer analysis methods are mainly used for the analysis of single herbs. There is no report on the thin layer identification analysis method for the whole formula of Anshen Dingzhi Pills. The general practice is to identify the herbs separately according to the thin layer identification conditions corresponding to the herbs such as ginseng, polygala, poria, poria and calamus in the formula of Anshen Dingzhi Pills. This is difficult to meet the requirements for the thin layer identification efficiency of Anshen Dingzhi Pills in actual production. At the same time, the ingredients of the compound formula of Anshen Dingzhi Pills are complex, and the thin layer analysis method of single herbs cannot accurately eliminate interference. It is urgent to develop a thin layer identification method for the whole formula of Anshen Dingzhi Pills.

[0004] The ingredients in Anshen Dingzhi Pills that can be identified by thin-layer chromatography include the main active ingredients of ginseng, including ginsenoside Rb1, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rf; Polygala tenuifolia, including index ingredients such as Polygala tenuifolia saponins; the index ingredients of Poria cocos and Poria cocos include triterpenoid compounds such as pachymic acid and dehydrogenated tert-butyl acid; and Acorus tatarinowii contains ingredients such as β-asarone. Due to the large structural differences of the above-mentioned index ingredients, the prior art basically uses specific identification methods and conditions to identify them separately, which greatly reduces the efficiency of the identification of the whole formula of Anshen Dingzhi Pills. How to achieve efficient identification of multiple index ingredients in the compound of Anshen Dingzhi Pills with simpler analysis conditions in the thin-layer analysis method is of great significance to the identification and quality control of the characteristic index ingredients of Anshen Dingzhi Pills. Summary of the invention

[0005] Based on the technical problems existing in the background technology, the present invention proposes a thin layer identification method for the index components in Anshen Dingzhi Pills.

[0006] The present invention provides a thin layer chromatography identification method for the index components in the Anshen Dingzhi Pill compound, comprising the following steps:

[0007] S1. Spotting: Pipette the Anshen Dingzhi Pills test solution, Anshen Dingzhi Pills negative test solution, control medicinal material solution and control substance solution and spot them on the silica gel G thin layer plate;

[0008] S2, developing: using a developing agent to develop the silica gel G thin layer plate, after development, taking out the silica gel G thin layer plate and drying it;

[0009] S3. Inspection: Spray the color developing solution on the dried silica gel G thin layer plate for color development inspection.

[0010] Anshen Dingzhi Pills include medicinal materials such as ginseng, Polygala tenuifolia, Poria cocos, Poria cocos and Acorus calamus; among them, the index components in ginseng include ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re and ginsenoside Rf; the index components in Polygala tenuifolia include Polygala tenuifolia saponins; the index components in Poria cocos, Poria cocos and Acorus calamus include pachymic acid, dehydrogenated tert-butyl acid and β-asarone.

[0011] Preferably, the thin layer chromatography identification method of the index component of ginseng in Anshen Dingzhi Pills comprises the following steps:

[0012] S1. Sample spotting: aspirate the Anshen Dingzhi Pills test solution 1, the Anshen Dingzhi Pills negative test solution 1, the control medicinal material solution 1 and the control substance solution and place them on a silica gel G thin layer plate 1;

[0013] S2, developing: using chloroform-methanol-water as a developing agent to develop the silica gel G thin layer plate, after development, taking out the silica gel G thin layer plate and drying it;

[0014] S3. Inspection: Spray sulfuric acid ethanol solution on the dried silica gel G thin layer plate for color development inspection.

[0015] More preferably, in the S1, the test solution of Anshen Dingzhi Pills is a solution of Anshen Dingzhi Pills with a mass volume ratio of 1 g / 10 mL dissolved by ultrasonic means of 75% ethanol; the negative test solution of Anshen Dingzhi Pills is a solution with a mass volume ratio of 2 g / 25 mL dissolved by ultrasonic means of 75% ethanol; the control medicinal material solution includes ginseng control medicinal material solutions with a mass volume ratio of 1 g / 50 mL each dissolved by ultrasonic means of 75% ethanol; the reference substance solution includes ginsenoside Rb1, ginsenoside Rg1, ginsenoside Re and ginsenoside Rf reference substance solutions with a mass volume ratio of 1 mg / 1 mL each dissolved by methanol.

[0016] More preferably, in the S1, the spotting volume ratio of Anshen Dingzhi Pills test solution 1, Anshen Dingzhi Pills negative test solution 1, control medicinal material solution 1, and reference substance solution 1 is 15:15:4:3.

[0017] More preferably, in the S2, the volume ratio of chloroform:methanol:water in the chloroform-methanol-water developing agent is 7:4:(0.5-1.5).

[0018] More preferably, in said S3, the mass percentage concentration of the sulfuric acid-ethanol mixed solution is 5%, wherein the volume ratio of the sulfuric acid solution to the ethanol solution is 1:19.

[0019] Preferably, the thin layer identification method of the indicative components of Polygala tenuifolia in Anshen Dingzhi Pills comprises the following steps:

[0020] S1. Spotting: aspirate the second Anshen Dingzhi Pills test solution, the second Anshen Dingzhi Pills negative test solution, the second reference medicinal material solution and the second reference substance solution and spot them on the second silica gel G thin layer plate;

[0021] S2, developing: using chloroform-methanol-water as a developing agent to develop the silica gel G thin layer plate, after development, taking out the silica gel G thin layer plate and drying it;

[0022] S3. Inspection: Spray sulfuric acid ethanol solution on the dried silica gel G thin layer plate for color development inspection.

[0023] More preferably, in the S1, the second test solution of Anshen Dingzhi Pills is a solution of Anshen Dingzhi Pills with a mass volume ratio of 1 g / 10 mL dissolved after ultrasonic extraction with 75% methanol; the second negative test solution of Anshen Dingzhi Pills is a solution with a mass volume ratio of 2 g / 25 mL dissolved after ultrasonic extraction with 75% methanol; the second control medicinal material solution includes a control medicinal material solution of Polygala tenuifolia with a mass volume ratio of 1 g / 50 mL dissolved after ultrasonic extraction with 75% methanol; the second reference substance solution includes a reference substance solution of Polygala tenuifolia saponins with a mass volume ratio of 1 mg / 1 mL dissolved in methanol.

[0024] More preferably, in the S1, the spotting volume ratio of the second Anshen Dingzhi Pills test solution, the second Anshen Dingzhi Pills negative test solution, the second control medicinal material solution, and the second reference substance solution is 20:20:20:8.

[0025] More preferably, in the S2, the volume ratio of chloroform:methanol:water in the chloroform-methanol-water developing agent is (7-7.5):4:(0.5-1.5).

[0026] More preferably, in said S3, the mass percentage concentration of the sulfuric acid-ethanol mixed solution is 5%, wherein the volume ratio of the sulfuric acid solution to the ethanol solution is 1:19.

[0027] Preferably, the thin layer identification method of the indicative components of Poria cocos, Poria cocos and Acorus tatarinowii in Anshen Dingzhi Pills comprises the following steps:

[0028] S1. Spotting: aspirate the Anshen Dingzhi Pills test solution 3, the Anshen Dingzhi Pills negative test solution 3, the control medicinal material solution 3 and the control substance solution 3 and spot them on the silica gel G thin layer plate 3;

[0029] S2, developing: using dichloromethane-ethyl acetate as a developing agent to develop the silica gel G thin layer plate, after development, taking out the silica gel G thin layer plate and drying it;

[0030] S3. Inspection: Spray a mixed solution of vanillin, sulfuric acid and ethanol on the dried silica gel G thin layer plate for color development inspection.

[0031] More preferably, in the S1, the Shendingzhi Pills test solution three is an Anshen Dingzhi Pills solution with a mass volume ratio of 1 g / 10 mL dissolved by ultrasonic dissolution in petroleum ether; the Anshen Dingzhi Pills negative test solution three is a solution with a mass volume ratio of (3-4.5) g / 50 mL dissolved by ultrasonic dissolution in petroleum ether; the control medicinal material solution three includes a solution with a mass volume ratio of (0.5-1) g / 50 mL dissolved by ultrasonic dissolution in petroleum ether; the reference substance solution three includes a dehydrogenated tert-butyl ether reference substance solution, a pachymic acid reference substance solution and a β-asarone reference substance solution dissolved in methanol with a mass volume ratio of 2 mg / 1 mL.

[0032] More preferably, in the S1, the volume ratio of the sample spots of Anshen Dingzhi Pills test solution 3, Anshen Dingzhi Pills negative test solution 3, control medicinal material solution 3, and reference substance solution 3 is 20:10:10:4.

[0033] More preferably, in said S2, the volume of dichloromethane-ethyl acetate in the dichloromethane-ethyl acetate developing agent is (4-4.5):(1-1.5).

[0034] More preferably, in said S3, the mass percentage concentration of the vanillin sulfuric acid ethanol mixed solution is 2%; wherein the volume ratio of the vanillin sulfuric acid solution to the ethanol solution is 1:9.

[0035] Preferably, in S3, the inspection condition is inspection under sunlight or under 365nm ultraviolet light.

[0036] The beneficial effects of the present invention are:

[0037] The present invention adopts a solution ultrasonic extraction method, and only needs to prepare three test sample solutions, and can simultaneously effectively identify 8 index components of ginsenoside Rb1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rf, fine leaf polygala saponin, dehydrogenated earth molar acid, pachymic acid and β-asarone in the Anshen Dingzhi Pill compound; the identification method of the present invention eliminates the influence of other components (dragon teeth, cinnabar) in the Anshen Dingzhi Pill on the thin layer identification, so that the chromatogram is clear and the separation is good. The present invention sets a negative test sample solution and a control medicinal material solution during the thin layer analysis, and the chromatogram comparison can be performed more clearly. It is found through the test results that the identification method proposed by the present invention has good applicability, the obtained chromatogram is clear, the separation is good, and the success rate of the thin layer identification of the Anshen Dingzhi Pill compound is significantly improved, which meets the requirements for the identification efficiency of the Anshen Dingzhi Pill in actual production. At the same time, the present invention detected the characteristic components of thin layer chromatography of Poria cocos and Acorus calamus (pachymic acid and β-asarone) in the 2020 edition of the "Chinese Pharmacopoeia", and also detected the characteristic components of thin layer chromatography of Poria cocos (dehydrogenated tert-butyl ether), which is also of great significance for the establishment of quality control indicators of Poria cocos, Poria cocos and Acorus calamus medicinal materials.

[0038] The present invention simultaneously detects the index components of tuckahoe, poria and calamus in the compound of Anshen Dingzhi Pills, namely pachymic acid, dehydrotrimoxan and β-asarone, under the condition of a test solution: petroleum ether is used as a solvent to prepare a dispersion solution of Anshen Dingzhi Pills with a mass volume ratio of 3 to 4.5 g / 50 mL, and after ultrasonic extraction, dichloromethane-ethyl acetate is used as a developing agent to develop the silica gel G thin layer plate, and the silica gel G thin layer plate is taken out after development and dried; a vanillin sulfuric acid ethanol mixed solution is sprayed on the dried silica gel G thin layer plate for color development inspection. As a result, in the chromatogram of the test sample of Anshen Dingzhi Pills, pachymic acid, dehydrotrimoxan and β-asarone show main spots of the same color at the corresponding positions of the chromatogram of the control medicinal material, and the separation effect is good.

[0039] The present invention uses a single identification method to simultaneously identify four ginsenosides in Anshen Dingzhi Pills, including ginsenoside Rb1, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rf. By controlling the experimental conditions, a chromatogram with clear chromatogram and good separation is obtained. As a result, in the chromatogram of the test sample of Anshen Dingzhi Pills, ginsenoside Rb1, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rf show main spots of the same color at positions corresponding to the chromatogram of the control ginseng medicinal material, and the separation effect is good. The extraction method and developing agent for thin-layer identification of a single ginseng medicinal material in the Chinese Pharmacopoeia are simpler and easier to operate, and effective thin-layer identification analysis of the compound Anshen Dingzhi Pills can be performed.

[0040] At the same time, the identification method of Polygala tenuifolia saponin, the indicative component of Polygala tenuifolia in the compound of Anshen Dingzhi Pills, is also simpler and easier to operate than the extraction method and developing agent for thin-layer identification of Polygala tenuifolia medicinal materials in the Chinese Pharmacopoeia. By controlling the experimental conditions, the chromatogram is clear and the separation is good. Results In the chromatogram of the test sample of Anshen Dingzhi Pills, Polygala tenuifolia saponin showed the same main spot of color at the corresponding position of the chromatogram of the control medicinal material, with good separation effect and high success rate of thin-layer identification, which can be used for thin-layer identification of Anshen Dingzhi Pills. The thin-layer chromatogram examined under sunlight is still clearly visible, which greatly improves the detection efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 a is a thin layer inspection diagram of Example 1 inspected under sunlight, Figure 1 b is a thin layer inspection diagram of Example 1 inspected under a UV lamp at 365 nm;

[0042] 1 is ginsenoside Rb1 reference solution; 2 is ginsenoside Re reference solution; 3 is ginsenoside Rf reference solution; 4 is ginsenoside Rg1 reference solution; 5 is ginsenoside Rb1, Re, Rf, Rg1 mixed reference solution; 6 is ginseng reference medicinal material solution; 7 is Anshen Dingzhi Pills test solution 1; 8 is Anshen Dingzhi Pills negative test solution 1.

[0043] Figure 2 This is a thin layer inspection diagram of Example 2 inspected under sunlight;

[0044] 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference medicinal material solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills.

[0045] Figure 3 This is a thin layer inspection diagram of Example 3 inspected under sunlight;

[0046] 1 is dehydrogenated taurine reference solution; 2 is pachymic acid reference solution; 3 is β-asarone reference solution; 4 is pachymarus reference medicinal material solution; 5 is calamus reference medicinal material solution; 6 is Anshen Dingzhi Pills test solution three; 7 is test solution three without pachymarus and poria; 8 is test solution three without calamus.

[0047] Figure 4 a is a thin layer inspection diagram of comparative example 1-1 inspected under sunlight, Figure 4 b is a thin layer inspection diagram of comparative example 1-1 inspected under a UV lamp at 365 nm;

[0048] 1 is ginsenoside Rb1 reference solution; 2 is ginsenoside Re reference solution; 3 is ginsenoside Rf reference solution; 4 is ginsenoside Rg1 reference solution; 5 is ginsenoside Rb1, Re, Rf, Rg1 mixed reference solution; 6 is ginseng reference medicinal material solution; 7 is Anshen Dingzhi Pills test solution 1; 8 is Anshen Dingzhi Pills negative test solution 1.

[0049] Figure 5 a is a thin layer inspection diagram of comparative example 1-2 inspected under sunlight, Figure 5 b is a thin layer inspection diagram of comparative example 1-2 inspected under a UV lamp at 365 nm;

[0050] 1 is ginsenoside Rb1 reference solution; 2 is ginsenoside Re reference solution; 3 is ginsenoside Rf reference solution; 4 is ginsenoside Rg1 reference solution; 5 is ginsenoside Rb1, Re, Rf, Rg1 mixed reference solution; 6 is ginseng reference medicinal material solution; 7 is Anshen Dingzhi Pills test solution 1; 8 is Anshen Dingzhi Pills negative test solution 1.

[0051] Figure 6 a is a thin layer inspection diagram of comparative example 1-3 inspected under sunlight, Figure 6 b is a thin layer inspection diagram of comparative example 1-3 inspected under a UV lamp at 365 nm;

[0052] 1 is ginsenoside Rb1 reference solution; 2 is ginsenoside Re reference solution; 3 is ginsenoside Rf reference solution; 4 is ginsenoside Rg1 reference solution; 5 is ginsenoside Rb1, Re, Rf, Rg1 mixed reference solution; 6 is ginseng reference medicinal material solution; 7 is Anshen Dingzhi Pills test solution 1; 8 is Anshen Dingzhi Pills negative test solution 1.

[0053] Figure 7 a is a thin layer inspection diagram of comparative examples 1-4 inspected under sunlight, Figure 7 b is a thin layer inspection diagram of comparative example 1-4 inspected under a UV lamp at 365 nm;

[0054] 1 is ginsenoside Rb1 reference solution; 2 is ginsenoside Re reference solution; 3 is ginsenoside Rf reference solution; 4 is ginsenoside Rg1 reference solution; 5 is ginsenoside Rb1, Re, Rf, Rg1 mixed reference solution; 6 is ginseng reference medicinal material solution; 7 is Anshen Dingzhi Pills test solution 1; 8 is Anshen Dingzhi Pills negative test solution 1.

[0055] Figure 8 This is a thin layer inspection diagram of Comparative Example 2-1 inspected under sunlight;

[0056] 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference medicinal material solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills.

[0057] Fig. 9 This is a thin layer inspection diagram of Comparative Example 2-2 inspected under sunlight;

[0058] 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference medicinal material solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills.

[0059] Fig.10 This is a thin layer inspection diagram of Comparative Example 2-3 inspected under sunlight;

[0060] 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference medicinal material solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills.

[0061] Fig.11 This is a thin layer inspection diagram of Comparative Examples 2-4 inspected under sunlight;

[0062] 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference medicinal material solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills.

[0063] Fig.12 This is a thin layer inspection diagram of Comparative Example 3-1 inspected under sunlight;

[0064] 1 is dehydrogenated taurine reference solution; 2 is pachymic acid reference solution; 3 is β-asarone reference solution; 4 is pachymarus reference medicinal material solution; 5 is calamus reference medicinal material solution; 6 is Anshen Dingzhi Pills test solution three; 7 is test solution three without pachymarus and poria; 8 is test solution three without calamus.

[0065] Fig.13 This is a thin layer inspection diagram of Comparative Example 3-2 inspected under sunlight;

[0066] 1 is dehydrogenated taurine reference solution; 2 is pachymic acid reference solution; 3 is β-asarone reference solution; 4 is pachymarus reference medicinal material solution; 5 is calamus reference medicinal material solution; 6 is Anshen Dingzhi Pills test solution three; 7 is test solution three without pachymarus and poria; 8 is test solution three without calamus.

[0067] Fig.14 This is a thin layer inspection diagram of Comparative Example 3-3 inspected under sunlight;

[0068] 1 is dehydrogenated taurine reference solution; 2 is pachymic acid reference solution; 3 is β-asarone reference solution; 4 is pachymarus reference medicinal material solution; 5 is calamus reference medicinal material solution; 6 is Anshen Dingzhi Pills test solution three; 7 is test solution three without pachymarus and poria; 8 is test solution three without calamus.

[0069] Fig.15 This is a thin layer inspection diagram of Comparative Example 3-4 inspected under sunlight;

[0070] 1 is dehydrogenated taurine reference solution; 2 is pachymic acid reference solution; 3 is β-asarone reference solution; 4 is pachymarus reference medicinal material solution; 5 is calamus reference medicinal material solution; 6 is Anshen Dingzhi Pills test solution three; 7 is test solution three without pachymarus and poria; 8 is test solution three without calamus. DETAILED DESCRIPTION

[0071] The technical solution of the present invention is described in detail through specific embodiments.

[0072] Unless otherwise specified, the materials, reagents, etc. used in the following examples and comparative examples can be obtained from commercial sources.

[0073] Example 1

[0074] This embodiment provides a thin layer chromatography identification method for ginsenoside Rb1, ginsenoside Rg1, ginsenoside Re and ginsenoside Rf in ginseng in Anshen Dingzhi Pills, comprising the following steps:

[0075] Preparation of Anshen Dingzhi Pills test solution 1: Take 5 g of Anshen Dingzhi Pills, grind them into powder, add 50 mL of 75% ethanol, ultrasonically treat (power 250 W, frequency 50 kHz) for 45 minutes, filter, evaporate the filtrate to dryness, add 1 mL of methanol to the residue to dissolve it, and use the Anshen Dingzhi Pills test solution 1.

[0076] Preparation of negative test solution 1 of Anshen Dingzhi Pills: take 4 g of pills made without ginseng, add 50 mL of 75% ethanol, ultrasonically treat (power 250 W, frequency 50 kHz) for 45 minutes, filter, evaporate the filtrate to dryness, add 1 mL of methanol to the residue to dissolve it, and use it as negative test solution 1 of Anshen Dingzhi Pills.

[0077] Preparation of control medicinal material solution 1: Take 1 g of ginseng control medicinal material and prepare it in the same way as Anshen Dingzhi Pills test solution 1 to obtain a 1 g / 50 mL ginseng control medicinal material solution.

[0078] Preparation of reference solution 1: Take ginsenoside Rb1 reference substance, ginsenoside Rg1 reference substance, ginsenoside Re reference substance and ginsenoside Rf reference substance and add methanol to make 1 mL of solution containing 1 mg of reference substance.

[0079] Spotting: For the thin layer chromatography (Chinese Pharmacopoeia 2020 Edition Part IV General Rules 0502) test, take 15 μL of the Anshen Dingzhi Pills test solution, 15 μL of the Anshen Dingzhi Pills negative test solution, 4 μL of the control medicinal material solution, and 3 μL of the reference solution (3 μL each of the ginsenoside Rb1 reference solution, ginsenoside Rg1 reference solution, ginsenoside Re, and ginsenoside Rf reference solution), and spot them on a silica gel G thin layer plate; use an automatic spotter to ensure that the spots are even and level during spotting;

[0080] Development: Use the lower layer solution of chloroform-methanol-water mixed solution with a volume ratio of 7:4:1 as the developing agent, place the spotted silica gel G thin layer plate in the developing agent vapor for pre-saturation, develop, take out and dry;

[0081] Inspection: Spray a 5% sulfuric acid ethanol solution (volume ratio of sulfuric acid solution to ethanol solution is 1:19) on the dried silica gel G thin layer plate, heat at 105°C until the spots are clearly colored, and inspect under sunlight and ultraviolet light (365nm).

[0082] The results are as follows Figure 1 As shown, 1 is ginsenoside Rb1 reference solution; 2 is ginsenoside Re reference solution; 3 is ginsenoside Rf reference solution; 4 is ginsenoside Rg1 reference solution; 5 is ginsenoside Rb1, Re, Rf, Rg1 mixed reference solution; 6 is ginseng reference medicinal material solution; 7 is Anshen Dingzhi Pills test solution 1; 8 is Anshen Dingzhi Pills negative test solution 1. It can be seen that in the Anshen Dingzhi Pills test sample chromatogram, the main spot of the same color appears at the corresponding position of the reference medicinal material chromatogram.

[0083] Example 2

[0084] This embodiment provides a thin layer chromatography identification method for polygala tenuifolia saponins in polygala tenuifolia in Anshen Dingzhi Pills, comprising the following steps:

[0085] Preparation of Anshen Dingzhi Pills test solution 2: Take about 5 g of Anshen Dingzhi Pills, grind them into powder, put them in a stoppered conical flask, accurately add 50 mL of 75% methanol, weigh the weight, ultrasonically treat (power 400 W, frequency 40 kHz) for 45 minutes, shake well, filter, evaporate to dryness, add 50 mL of water-saturated n-butanol and shake to extract, let stand to separate the layers, take the supernatant, evaporate to dryness, add 1 mL of methanol to the residue to dissolve it, and obtain Anshen Dingzhi Pills test solution 2.

[0086] Preparation of negative test solution 2 of Anshen Dingzhi Pills: Take about 4 g of pills made from Polygala tenuifolia, grind them into powder, put them in a stoppered conical flask, accurately add 50 mL of 75% methanol, weigh the weight, ultrasonically treat (power 400 W, frequency 40 kHz) for 45 minutes, shake well, filter, evaporate to dryness, add 50 mL of water-saturated n-butanol and shake to extract, let stand to separate the layers, take the supernatant, evaporate to dryness, add 1 mL of methanol to the residue to dissolve it, and the negative test solution 2 of Anshen Dingzhi Pills is obtained.

[0087] Preparation of control medicinal solution 2: Take 1 g of Polygala tenuifolia control medicinal material and prepare it in the same manner as Anshen Dingzhi Pills test solution 2 to obtain 1 g / 50 mL of Polygala tenuifolia control medicinal solution.

[0088] Preparation of reference solution 2: Take the Polygala tenuifolia saponin reference standard and add methanol to make 1 mL of a solution containing 1 mg, the Polygala tenuifolia saponin reference standard solution.

[0089] Spotting: Thin layer chromatography (Chinese Pharmacopoeia 2020 Edition Part IV General Rules 0502) test, take 20μL of Anshen Dingzhi Pills test solution 2, 20μL of Anshen Dingzhi Pills negative test solution 2, 20μL of control medicinal material solution 2, and 8μL of reference solution 2, and spot them on silica gel G thin layer plate 2 respectively; use an automatic spotter to ensure that the spots are even and level during spotting;

[0090] Development: Use the lower layer solution of chloroform-methanol-water mixed solution with a volume ratio of 7:4:1 as the developing agent to place the spotted silica gel G thin layer plate in the developing agent vapor for pre-saturation, develop, take out and dry;

[0091] Inspection: Spray a 5% sulfuric acid ethanol solution (volume ratio of sulfuric acid solution to ethanol solution is 1:19) on the dried silica gel G thin layer plate 2, and heat at 105°C until the spots are clearly colored.

[0092] The results are as follows Figure 2 As shown, 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills. It can be seen that in the chromatogram of the test sample of Anshen Dingzhi Pills, the main spot of the same color appears at the corresponding position of the chromatogram of the reference medicinal material, and the separation effect is good.

[0093] Example 3

[0094] This embodiment provides a thin layer chromatography identification method for dehydrogenated turmeric acid and pachymic acid in Poria cocos and Poria cocos in Anshen Dingzhi Pills and β-asarone in Acorus tatarinowii, comprising the following steps:

[0095] Preparation of Anshen Dingzhi Pills test solution three: Take 5 g of Anshen Dingzhi Pills, grind them into powder, add 50 mL of petroleum ether, ultrasonically treat (power 250 W, frequency 50 kHz) for 30 minutes, filter, evaporate the filtrate to dryness, add 1 mL of methanol to dissolve the residue, and use the Anshen Dingzhi Pills test solution three.

[0096] Preparation of negative test solution three for Anshen Dingzhi Pills: take 3 g of pills made without Poria cocos and Poria spirit and 4.5 g of pills made without Acorus tatarinowii, grind them into powder, add 50 mL of petroleum ether to each, treat with ultrasound (power 250 W, frequency 50 kHz) for 30 min, filter, evaporate the filtrate to dryness, add 1 mL of methanol to dissolve the residues, and use them as negative test solution three for Anshen Dingzhi Pills, namely, test solution three without Poria cocos and Poria spirit and test solution three without Acorus tatarinowii.

[0097] Preparation of control medicinal solution three: Take 0.5g of Acorus tatarinowii control medicinal material and 1g of Poria cocos control medicinal material respectively, and prepare them in the same way as Anshen Dingzhi Pills test solution three to obtain 0.5g / 50mL Acorus tatarinowii control medicinal material solution and 1g / 50mL Poria cocos control medicinal material solution.

[0098] Preparation of reference solution three: Take dehydrogenated tert-butyl ester reference standard, pachymic acid reference standard, β-asarone reference standard, add methanol to make 1 mL of solution containing 2 mg each, to obtain dehydrogenated tert-butyl ester reference solution, pachymic acid reference solution and β-asarone reference solution.

[0099] Spotting: Thin layer chromatography (Chinese Pharmacopoeia 2020 Edition Part IV General Rules 0502) test, take 20μL of Anshen Dingzhi Pills test solution 3, 10μL of Anshen Dingzhi Pills negative test solution 3 (10μL of each of the two negative test solutions), 10μL of control medicinal material solution 3 (10μL of Poria control medicinal material solution and Acorus tatarinowii control medicinal material solution), 4μL of reference solution 3 (4μL of dehydrogenated tert-butyl acid reference solution, tuckahoe acid reference solution and β-asarone reference solution), and spot them on silica gel G thin layer plate 3; use an automatic spotter to ensure that the spots are even and level during spotting;

[0100] Development: Use a mixed solution of dichloromethane and ethyl acetate with a volume ratio of 4:1 as the developing agent, place the spotted silica gel G thin layer plate in the developing agent vapor for pre-saturation, develop, take out, and dry;

[0101] Inspection: Spray a mixed solution of 2% vanillin sulfate ethanol (the volume of vanillin sulfate solution and ethanol is 1:9) on the dried silica gel G thin layer plate 3, and heat at 105°C until the spots are clearly colored.

[0102] The results are as follows Figure 3As shown, 1 is dehydrogenated taurine acid reference solution; 2 is tuckahoe acid reference solution; 3 is β-asarone reference solution; 4 is tuckahoe reference medicinal material solution; 5 is calamus tatarinowii reference medicinal material solution; 6 is Anshen Dingzhi Pills test solution 3; 7 is Poria cocos Poria test solution 3 without Poria cocos; 8 is calamus tatarinowii test solution 3. It can be seen that in the chromatogram of the Anshen Dingzhi Pills test sample, the main spot of the same color appears at the corresponding position of the chromatogram of the reference medicinal material, and the separation effect is good.

[0103] Comparative Example 1-1

[0104] The same method as in Example 1 was used to perform thin layer identification on the indicative components in ginseng in Anshen Dingzhi Pills, except that during the development process, the developing agent chloroform-methanol-water (7:4:1) was replaced with chloroform-ethyl acetate-methanol-water (15:40:22:10).

[0105] The results are as follows Figure 4 As shown, in the chromatogram of the test sample of Anshen Dingzhi Pills, a main spot of the same color appears at the corresponding position in the chromatogram of the control medicinal material, but compared with the developing agent chloroform-methanol-water (7:4:1), the spot is slightly blurred and the separation is not good enough, but it can still be distinguished.

[0106] Comparative Example 1-2

[0107] The indicative components in ginseng in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 1, except that during the development process, the developing agent chloroform-methanol-water (7:4:1) was replaced with chloroform-methanol-water (7:3:1).

[0108] The results are as follows Figure 5 The results show that in the chromatogram of the Anshen Dingzhi Pills test sample, the main spot of the same color appears at the corresponding position of the chromatogram of the control medicinal material. However, compared with the developing solvent chloroform-methanol-water (7:4:1), the spot is slightly blurred and the separation is not good enough, but it can still be distinguished.

[0109] Comparative Examples 1-3

[0110] The indicator components in ginseng in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 1, except that, during the spotting process, 10 μL of Anshen Dingzhi Pills test solution, 10 μL of Anshen Dingzhi Pills negative test solution, 2 μL of control medicinal material solution, and 2 μL of reference solution (2 μL each of ginsenoside Rb1 reference solution, ginsenoside Rg1 reference solution, ginsenoside Re, and ginsenoside Rf reference solution) were aspirated.

[0111] The results are as follows Figure 6As shown, 1 is ginsenoside Rb1 reference solution; 2 is ginsenoside Re reference solution; 3 is ginsenoside Rf reference solution; 4 is ginsenoside Rg1 reference solution; 5 is ginsenoside Rb1, Re, Rf, Rg1 mixed reference solution; 6 is ginseng reference medicinal material solution; 7 is Anshen Dingzhi Pills test solution 1; 8 is Anshen Dingzhi Pills negative test solution 1. It can be seen that in the Anshen Dingzhi Pills test sample chromatogram, the main spot of the same color appears at the corresponding position of the reference medicinal material chromatogram. It can be seen that at the position corresponding to the chromatogram of the control medicinal material, the main colored spot is blurred, compared with the absorption of Anshen Dingzhi Pills test solution - 15 μL, Anshen Dingzhi Pills negative test solution - 15 μL, control medicinal material solution - 4 μL, reference solution - 3 μL (3 μL each of ginsenoside Rb1 reference solution, ginsenoside Rg1 reference solution, ginsenoside Re, and ginsenoside Rf reference solution).

[0112] Comparative Examples 1-4

[0113] The indicative components in ginseng in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 1, except that during the development process, the developing agent chloroform-methanol-water (7:4:1) was replaced with chloroform-methanol-water (13:7:2).

[0114] The results are as follows Figure 7 As shown, after the developing agent chloroform-methanol-water (7:4:1) was replaced with chloroform-methanol-water (13:7:2), the spots were blurred.

[0115] Comparative Example 2-1

[0116] The indicative components in Polygala tenuifolia in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 2, except that in the developing step, the developing agent was a mixed solution of chloroform-methanol-water in a volume ratio of 6:3:0.5.

[0117] The results are as follows Figure 8 As shown, 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills. It can be seen that the development effect is not ideal, and the main spot of color is not clear at the position corresponding to the chromatogram of the reference medicinal material.

[0118] Comparative Example 2-2

[0119] The indicative components in Polygala tenuifolia in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 2, except that in the developing step, the developing agent was a mixed solution of chloroform-methanol-water in a volume ratio of 6:4:1.

[0120] The results are as follows Fig. 9 As shown, 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills. It can be seen that the development effect is not ideal, and the main spot of color is not clear at the position corresponding to the chromatogram of the reference medicinal material.

[0121] Comparative Examples 2-3

[0122] The same method as in Example 2 was used to perform thin layer identification on the indicative components in Polygala tenuifolia in Anshen Dingzhi Pills, except that in the spotting step, 15 μL of Anshen Dingzhi Pills test solution, 15 μL of Anshen Dingzhi Pills negative test solution, 15 μL of control medicinal material solution, and 6 μL of control substance solution were aspirated.

[0123] The results are as follows Fig.10 As shown, 1 is the reference solution of Polygala tenuifolia saponins; 2 is the reference solution of Polygala tenuifolia; 3 is the second test solution of Anshen Dingzhi Pills; 4 is the second negative test solution of Anshen Dingzhi Pills. It can be seen that compared with the absorption of 20 μL of the second test solution of Anshen Dingzhi Pills, 20 μL of the second negative test solution of Anshen Dingzhi Pills, 20 μL of the second control solution of the medicinal material, and 8 μL of the second reference solution, the main spot of color development is not clear at the position corresponding to the chromatogram of the control medicinal material.

[0124] Comparative Examples 2-4

[0125] The indicative components in Polygala tenuifolia in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 2, except that, in the developing step, the developing agent was a mixed solution of chloroform-ethyl acetate-methanol-water in a volume ratio of 15:40:22:10.

[0126] The results are as follows Fig.11 As shown, when the developing agent is a mixed solution of chloroform-ethyl acetate-methanol-water with a mixed volume ratio of 15:40:22:10, although the spots are clearly colored, interference occurs with the negative test sample.

[0127] Comparative Example 3-1

[0128] The index components in Poria cocos, Poria cocos and Acorus tatarinowii in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 3, except that in the developing step, the developing agent was a mixed solution of toluene-ethyl acetate-formic acid in a mixing volume ratio of 20:5:0.5;

[0129] The results are as follows Fig.12As shown, 1 is the dehydrogenated taurine acid reference solution; 2 is the tuckahoe acid reference solution; 3 is the β-asarone reference solution; 4 is the tuckahoe reference medicinal material solution; 5 is the calamus tatarinowii reference medicinal material solution; 6 is the Anshen Dingzhi Pill test solution 3; 7 is the tuckahoe tatarinowii test solution 3 without tuckahoe tatarinowii; 8 is the calamus tatarinowii test solution 3. It can be seen that at the corresponding position of the reference medicinal material chromatogram, the main spot of the same color appears, but the spot color is not clear enough, and the separation effect of the β-asarone band in the test sample is not good.

[0130] Comparative Example 3-2

[0131] The index components in Poria cocos, Poria cocos and Acorus tatarinowii in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 3, except that in the developing step, the developing agent was a mixed solution of toluene-ethyl acetate-formic acid with a mixing volume ratio of 22:5:1;

[0132] The results are as follows Fig.13 As shown, 1 is dehydrogenated taurine acid reference solution; 2 is tuckahoe acid reference solution; 3 is β-asarone reference solution; 4 is tuckahoe reference medicinal material solution; 5 is calamus tatarinowii reference medicinal material solution; 6 is Anshen Dingzhi Pills test solution 3; 7 is tuckahoe fushen test solution 3 without tuckahoe fushen; 8 is calamus tatarinowii test solution 3. It can be seen that at the position corresponding to the reference medicinal material chromatogram, the main spot of the same color appears, and the separation effect is good, but the spot color is not clear enough.

[0133] Comparative Example 3-3

[0134] The same method as in Example 3 was used to perform thin layer identification on the indicator components in Poria, Poria and Acorus tatarinowii in Anshen Dingzhi Pills, with the difference that, in the spotting step, 15 μL of three Anshen Dingzhi Pills test solution, 15 μL of three Anshen Dingzhi Pills negative test solution (15 μL of each of the two negative test solutions), 15 μL of three control medicinal material solutions (15 μL of each Poria control medicinal material solution and Acorus tatarinowii control medicinal material solution), and 6 μL of three reference substance solutions (6 μL of each of dehydrogenated tert-butyl ether reference substance solution, pachymic acid reference substance solution and β-asarone reference substance solution) were aspirated.

[0135] The results are as follows Fig.14As shown, 1 is dehydrogenated tert-butyl acid reference solution; 2 is tuckahoe acid reference solution; 3 is β-asarone reference solution; 4 is tuckahoe control medicinal material solution; 5 is calamus tatarinowii control medicinal material solution; 6 is Anshen Dingzhi Pill test solution 3; 7 is tuckahoe tatarinowii test solution 3 without tuckahoe tatarinowii; 8 is calamus tatarinowii test solution 3 without tuckahoe tatarinowii. It can be seen that compared with the absorption of 20 μL of Anshen Dingzhi Pill test solution 2, 10 μL of Anshen Dingzhi Pill negative test solution 2 (10 μL of each of the two negative test solutions), 10 μL of control medicinal material solution 3 (10 μL of each of tuckahoe control medicinal material solution and calamus tatarinowii control medicinal material solution), and 4 μL of control solution 3 (4 μL of each of dehydrogenated tert-butyl acid reference solution, tuckahoe acid reference solution, and β-asarone reference solution), the main spots of the same color are shown at the corresponding positions of the control medicinal material chromatogram, but the spot color is not clear enough.

[0136] Comparative Examples 3-4

[0137] The index components in Poria cocos, Poria cocos and Acorus tatarinowii in Anshen Dingzhi Pills were identified by thin layer chromatography in the same manner as in Example 3, except that in the developing step, the developing agent was a mixed solution of dichloromethane-ethyl acetate-formic acid with a mixing volume ratio of 12:3:0.5;

[0138] The results are as follows Fig.15 As shown, when the developing solvent is a mixed solution of dichloromethane-ethyl acetate-formic acid with a mixing volume ratio of 12:3:0.5, the separation of β-asarone in the test sample is poor and the color development is relatively vague.

[0139] In summary, the thin layer chromatography identification method for the index components in Anshen Dingzhi Pills proposed in the present invention has good applicability, the obtained chromatogram is clear, and the separation is good, which significantly improves the success rate of thin layer chromatography identification of Anshen Dingzhi Pills and meets the requirements for the identification efficiency of Anshen Dingzhi Pills in actual production.

[0140] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A thin layer identification method for the index components in Anshen Dingzhi Pills, characterized in that: The following steps are involved: S1. Spotting: Pipette the Anshen Dingzhi Pills test solution, Anshen Dingzhi Pills negative test solution, control medicinal material solution and control substance solution and spot them on the silica gel G thin layer plate; S2, developing: using a developing agent to develop the silica gel G thin layer plate, taking out the silica gel G thin layer plate after development and drying it; S3, inspection: spray the color developing solution on the dried silica gel G thin layer plate for color development inspection; The Chinese medicinal materials identified by thin layer chromatography in Anshen Dingzhi Pills include ginseng, Polygala tenuifolia, Poria cocos, Poria cocos and Acorus tatarinowii; among them, the index components in ginseng include ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re and ginsenoside Rf; the index components in Polygala tenuifolia include Polygala tenuifolia saponin; the index components in Poria cocos, Poria cocos and Acorus tatarinowii include pachymic acid, dehydrogenated tert-butyl acid and β-asarone; In the above-mentioned S3, the inspection condition is inspection under sunlight or under 365nm ultraviolet light.

2. The thin layer identification method according to claim 1, characterized in that: The thin-layer identification method of the index component of ginseng in Anshen Dingzhi Pills comprises the following steps: S1. Sample spotting: aspirate the Anshen Dingzhi Pills test solution 1, the Anshen Dingzhi Pills negative test solution 1, the control medicinal material solution 1 and the control substance solution and place them on a silica gel G thin layer plate 1; S2, developing: using chloroform-methanol-water as a developing agent to develop the silica gel G thin layer plate, after development, taking out the silica gel G thin layer plate and drying it; S3. Inspection: Spray sulfuric acid ethanol solution on the dried silica gel G thin layer plate for color development inspection.

3. The thin layer identification method according to claim 2, characterized in that: In the described S1, the test solution of Anshen Dingzhi Pills is a solution of Anshen Dingzhi Pills with a mass volume ratio of 1 g / 10 mL dissolved by ultrasonication in 75% ethanol; the negative test solution of Anshen Dingzhi Pills is a solution with a mass volume ratio of 2 g / 25 mL dissolved by ultrasonication in 75% ethanol; the control medicinal material solution includes ginseng control medicinal material solutions with a mass volume ratio of 1 g / 50 mL each dissolved by ultrasonication in 75% ethanol; the reference substance solution includes ginsenoside Rb1, ginsenoside Rg1, ginsenoside Re and ginsenoside Rf reference substance solutions with a mass volume ratio of 1 mg / 1 mL each dissolved by methanol.

4. The thin layer identification method according to claim 2, characterized in that: In the described S1, the volume ratio of the sample volume of Anshen Dingzhi Pills test solution 1, Anshen Dingzhi Pills negative test solution 1, control medicinal material solution 1, and reference solution 1 is 15:15:4:3; in the described S2, the volume ratio of chloroform:methanol:water in the chloroform-methanol-water developing agent is 7:4:(0.5-1.5); in the described S3, the mass percentage concentration in the sulfuric acid-ethanol mixed solution is 5%, wherein the volume ratio of the sulfuric acid solution to the ethanol solution is 1:

19.

5. The thin layer identification method according to claim 1, characterized in that: The thin layer identification method of the index components of Polygala tenuifolia in Anshen Dingzhi Pills comprises the following steps: S1. Spotting: aspirate the second Anshen Dingzhi Pills test solution, the second Anshen Dingzhi Pills negative test solution, the second reference medicinal material solution and the second reference substance solution and spot them on the second silica gel G thin layer plate; S2, developing: using chloroform-methanol-water as a developing agent to develop the silica gel G thin layer plate, after development, taking out the silica gel G thin layer plate and drying it; S3. Inspection: Spray sulfuric acid ethanol solution on the dried silica gel G thin layer plate for color development inspection.

6. The thin layer identification method according to claim 5, characterized in that: In the S1, the second test solution of Anshen Dingzhi Pills is a solution of Anshen Dingzhi Pills with a mass volume ratio of 1g / 10mL dissolved after ultrasonic extraction with 75% methanol; the second negative test solution of Anshen Dingzhi Pills is a solution with a mass volume ratio of 2g / 25mL dissolved after ultrasonic extraction with 75% methanol; the second control medicinal material solution includes a control medicinal material solution of Polygala tenuifolia with a mass volume ratio of 1g / 50mL dissolved after ultrasonic extraction with 75% methanol; the second reference substance solution includes a reference substance solution of Polygala tenuifolia saponins with a mass volume ratio of 1mg / 1mL dissolved in methanol.

7. The thin layer identification method according to claim 5, characterized in that: In the described S1, the volume ratio of the sample volume of Anshen Dingzhi Pills test solution 2, Anshen Dingzhi Pills negative test solution 2, control medicinal material solution 2, and reference substance solution 2 is 20:20:20:8; in the described S2, the volume ratio of chloroform:methanol:water in the chloroform-methanol-water developing agent is (7-7.5):4:(0.5-1.5); in the described S3, the mass percentage concentration in the sulfuric acid-ethanol mixed solution is 5%, wherein the volume ratio of the sulfuric acid solution to the ethanol solution is 1:

19.

8. The thin layer identification method according to claim 1, characterized in that: The thin layer identification method of the index components of Poria cocos, Poria cocos and Acorus tatarinowii in Anshen Dingzhi Pills comprises the following steps: S1. Spotting: aspirate the Anshen Dingzhi Pills test solution 3, the Anshen Dingzhi Pills negative test solution 3, the control medicinal material solution 3 and the control substance solution 3 and spot them on the silica gel G thin layer plate 3; S2, developing: using dichloromethane-ethyl acetate as a developing agent to develop the silica gel G thin layer plate, after development, taking out the silica gel G thin layer plate and drying it; S3. Inspection: Spray a mixed solution of vanillin, sulfuric acid and ethanol on the dried silica gel G thin layer plate for color development inspection.

9. The thin layer identification method according to claim 8, characterized in that: In the S1, the Shendingzhi Pills test solution three is an Anshen Dingzhi Pills solution with a mass volume ratio of 1 g / 10 mL dissolved by ultrasonic means of petroleum ether; the Anshen Dingzhi Pills negative test solution three is a solution with a mass volume ratio of (3-4.5) g / 50 mL dissolved by ultrasonic means of petroleum ether; the control medicinal material solution three includes a solution with a mass volume ratio of (0.5-1) g / 50 mL dissolved by ultrasonic means of petroleum ether; the reference substance solution three includes a dehydrogenated tert-butyl ether reference substance solution, a pachymic acid reference substance solution and a β-asarone reference substance solution dissolved by methanol with a mass volume ratio of 2 mg / 1 mL.

10. The thin layer identification method according to claim 8, characterized in that: In the described S1, the volume ratio of the sample volume of Anshen Dingzhi Pills test solution 3, Anshen Dingzhi Pills negative test solution 3, control medicinal material solution 3, and reference substance solution 3 is 20:10:10:4; in the described S2, the volume of dichloromethane-ethyl acetate in the dichloromethane-ethyl acetate developing agent is (4-4.5):(1-1.5); in the described S3, the mass percentage concentration of the vanillin sulfuric acid ethanol mixed solution is 2%; wherein the volume ratio of the vanillin sulfuric acid solution to the ethanol solution is 1:9.

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