A thin layer identification method for granules with different original formulas of Sichuan tung bark

Abstract

The present invention provides a thin layer chromatography identification method for granules with different origins of Sichuan tung bark, comprising the following steps: A) taking a sample to be tested, dissolving it with a solvent to obtain a test solution; the sample to be tested is a granule of Chinese catalpa ovata or a granule of Chinese catalpa ovata; B) taking a control medicinal material, dissolving it with a solvent to obtain a control medicinal material solution; the control medicinal material is a control medicinal material of Chinese catalpa ovata or a control medicinal material of Chinese catalpa ovata; C) subjecting the test solution and the control medicinal material solution to thin layer chromatography, using a silica gel G thin layer plate and a developing solvent of petroleum ether-chloroform-ethanol-glacial acetic acid; D) examining the sample under ultraviolet light; if a blue fluorescent spot is present at an Rf value of 0.235±10% in the chromatogram of the sample, the sample is Chinese catalpa ovata; if no blue fluorescent spot is present at an Rf value of 0.235±10%, the sample is Chinese catalpa ovata. This method has good stability, high precision, good reproducibility, is convenient, and is easy to master.

Description

Technical Field

[0001] The invention belongs to the technical field of analysis and detection, and in particular relates to a thin layer identification method for granules with different original formulas of Sichuan tung bark. Background Art

[0002] Chuantong bark is the dried bark of Kalopanax septemlobus (Thunb.) Koidz. or Kalopanax septemlobus var. margnificus (Zabel) Hand-Mazz., both of the Araliaceae family. It has the effects of dispelling rheumatism, unblocking meridians, and relieving pain. [1] . Used for rheumatoid arthritis, pain in the waist and knees; used externally for traumatic injuries.

[0003] The "Technical Requirements for Quality Control and Standardization of Traditional Chinese Medicine Formula Granules" published by the State Food and Drug Administration stipulates that the raw medicinal materials used in traditional Chinese medicine formula granules should be of fixed origin. Property identification is an effective method for distinguishing the two. Formula granules are made from medicinal materials through processing, decoction, concentration, drying, mixing, and granulation. Decoction will destroy the original properties of the medicinal materials, and property identification methods are not applicable to the identification of formula granules. No relevant literature has been reported on the basic principle and thin layer identification methods of different bases of Sichuan tung bark. Currently, there is no effective method to distinguish between Sichuan tung bark (Catharanthus spinulosa) formula granules and Sichuan tung bark (Catharanthus pubescens) formula granules. Therefore, it is urgent to establish a method that can distinguish Sichuan tung bark (Catharanthus spinulosa) formula granules from Sichuan tung bark (Catharanthus pubescens) formula granules. Summary of the Invention

[0004] The purpose of the present invention is to provide a thin layer identification method for particles with different base formulas of Chuantong bark. The method of the present invention has good stability, high precision, good reproducibility, is convenient and easy to master.

[0005] The terms "include," "comprising," and "having" are used interchangeably herein and are intended to indicate the inclusiveness of a solution, meaning that other elements may be present in addition to the listed elements. It should also be understood that the use of "include," "comprising," and "having" in this document also provides a "consisting of" solution.

[0006] In this application, the term "and / or" describes the association relationship between associated objects, indicating that three relationships may exist. For example, A and / or B can mean: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural.

[0007] In this application, "at least one" means one or more, and "more than one" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items.

[0008] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the following processes does not mean the order of execution. Some or all of the steps can be executed in parallel or sequentially. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0009] The present invention aims to establish a thin-layer chromatography method for identifying the formula granules of Chuantong bark (catalpa asiatica) and Chuantong bark (hairy-leaved catalpa asiatica). By observing whether there are fluorescent spots within a certain ratio shift value range on the thin-layer chromatogram of the test sample, the method is used as an identification point to quickly and effectively identify the formula granules of Chuantong bark (catalpa asiatica) and Chuantong bark (hairy-leaved catalpa asiatica).

[0010] The present invention provides a thin layer identification method for granules of different original formulas of Sichuan tung bark, comprising the following steps:

[0011] A) taking a sample to be tested and dissolving it in a solvent to obtain a test solution; the sample to be tested is a granule of Castanopsis arvense or a granule of Castanopsis pubescens;

[0012] B) taking a control medicinal material and dissolving it with a solvent to obtain a control medicinal material solution; the control medicinal material is a Catalpa oleracea control medicinal material or a Catalpa oleracea hairy-leaved control medicinal material;

[0013] C) The test solution and the control medicinal material solution were subjected to thin layer chromatography using a silica gel G thin layer plate and a developing solvent of petroleum ether-chloroform-ethanol-glacial acetic acid;

[0014] D) Check under ultraviolet light. If a blue fluorescent spot is found at an Rf value of 0.235±10% in the chromatogram of the test sample, it is Catalpa ovata. If no blue fluorescent spot is found at an Rf value of 0.235±10%, it is Catalpa ovata.

[0015] The present invention preferably prepares the test solution according to the following steps:

[0016] Take the sample to be tested, add water and hydrochloric acid, heat under reflux for hydrolysis, extract with n-butanol, and combine the extracts;

[0017] In the present invention, the sample to be tested is the formula granules of Chuantong bark (Castanopsis pilosula) or the formula granules of Chuantong bark (Castanopsis pilosula).

[0018] In the present invention, the ratio of the mass of the sample to be tested to the volume of water is preferably 1 g: (10-25) mL, more preferably 1 g: (15-20) mL; the ratio of the mass of the sample to be tested to the volume of hydrochloric acid is preferably 1 g: (3-5) mL, more preferably 1 g: (3-4) mL.

[0019] In the present invention, the hydrolysis time is preferably 1 to 3 hours, more preferably 2 hours.

[0020] After the extract is evaporated to dryness, the residue is dissolved in water and passed through a neutral alumina column. First, elution is performed with 50 to 60 mL of water, the water eluate is discarded, and then elution is performed with 25 to 30 mL of ethanol. The ethanol eluate is collected and the solvent is recovered to dryness.

[0021] Add 1-2 mL of methanol to the residue and sonicate, take the filtrate to obtain the test solution.

[0022] In the present invention, the ultrasonic time is preferably 25 to 35 minutes, more preferably 30 minutes.

[0023] In the present invention, the control medicinal material solution is preferably prepared according to the following steps:

[0024] Take the control medicinal material, add water and hydrochloric acid, heat under reflux for hydrolysis, extract with n-butanol, and combine the extracts;

[0025] In the present invention, the control medicinal material is the Chuantong bark (Calotaxus arborescens) control medicinal material or the Chuantong bark (Calotaxus arborescens) control medicinal material.

[0026] In the present invention, the ratio of the mass of the control medicinal material to the volume of water is preferably 1 g: (10-25) mL, more preferably 1 g: (15-20) mL; the ratio of the mass of the control medicinal material to the volume of hydrochloric acid is preferably 1 g: (3-5) mL, more preferably 1 g: (3-4) mL.

[0027] In the present invention, the hydrolysis time is preferably 1 to 3 hours, more preferably 2 hours.

[0028] After the extract is evaporated to dryness, the residue is dissolved in water and passed through a neutral alumina column. First, elution is performed with 50 to 60 mL of water, the water eluate is discarded, and then elution is performed with 25 to 30 mL of ethanol. The ethanol eluate is collected and the solvent is recovered to dryness.

[0029] Add 1-2 mL of methanol to the residue and sonicate, take the filtrate to obtain the test solution.

[0030] In the present invention, the ultrasonic time is preferably 25 to 35 minutes, more preferably 30 minutes.

[0031] After obtaining the test solution and the control medicinal material solution, the present invention performs thin layer chromatography on the test solution and the control medicinal material solution. The specific steps are as follows:

[0032] The present invention preferably tests according to the thin layer chromatography method (General Rules 0502 of the 2020 edition of the Chinese Pharmacopoeia), and the test sample solution and the control medicinal material solution are respectively spotted on the same silica gel G thin layer plate, developed with a developing agent, taken out, dried, sprayed with 10% sulfuric acid ethanol, heated at 105°C until the spots are colored, washed, and placed under ultraviolet light for inspection.

[0033] In the present invention, the developing agent is preferably petroleum ether-chloroform-ethanol-glacial acetic acid, the boiling range of the petroleum ether is preferably 30-60°C, and the mass ratio of the petroleum ether, chloroform, ethanol and glacial acetic acid is (13-17):(4.5-7.5):(0.8-1.2):0.3, more preferably (14-16):(5-7):(0.9-1.1):0.3, and most preferably 15:6:1:0.3.

[0034] In the present invention, the spotting volume of the test solution and the control medicinal material solution is preferably 3 to 5 μL.

[0035] In the present invention, the wavelength of the ultraviolet lamp is preferably 365 nm.

[0036] In the present invention, after ultraviolet light inspection, if there is a blue fluorescent spot at the Rf value of 0.235±10% in the chromatogram of the test sample, it is Catalpa ovata; if there is no blue fluorescent spot at the Rf value of 0.235±10%, it is Catalpa ovata.

[0037] In the present invention, the Rf value is preferably 0.235±10%, more preferably 0.217-0.245, such as 0.217, 0.220, 0.223, 0.225, 0.228, 0.230, 0.232, 0.235, 0.238, 0.240, 0.242, 0.245.

[0038] In the present invention, the inspection temperature is preferably 4-30° C., and the method of the present invention has good adaptability to different temperatures.

[0039] In the present invention, the humidity of the inspection is preferably 32% rh to 75% rh, and the method of the present invention has good adaptability to different humidity levels.

[0040] The invention provides a thin layer chromatography identification method for granules with different origins of Sichuan tung bark, comprising the following steps: A) taking a sample to be tested, dissolving it with a solvent, and obtaining a test solution; the sample to be tested is a Chinese catalpa vine formula granule or a Chinese catalpa vine hairy-leaf formula granule; B) taking a control medicinal material, dissolving it with a solvent, and obtaining a control medicinal material solution; the control medicinal material is a Chinese catalpa vine control medicinal material or a Chinese catalpa vine hairy-leaf control medicinal material; C) subjecting the test solution and the control medicinal material solution to thin layer chromatography detection, wherein the thin layer plate is a silica gel G thin layer plate and the developing solvent is petroleum ether-chloroform-ethanol-glacial acetic acid; D) inspecting under an ultraviolet lamp, and if a blue fluorescent spot is present at an Rf value of 0.235±10% in the chromatogram of the test sample, the sample is Chinese catalpa vine hairy-leaf; and if no blue fluorescent spot is present at an Rf value of 0.235±10%, the sample is Chinese catalpa vine hairy-leaf.

[0041] The thin layer identification method of the present invention compares the thin layer chromatography of the test sample with the thin layer chromatography of the control medicinal material Chuantong bark (Calotaxus thornii). If the spots are consistent with those of the control medicinal material, it is Chuantong bark (Calotaxus thornii) granules; if the blue fluorescent spots are at the same position as the control medicinal material at the ratio shift value of 0.217 to 0.245, it is Chuantong bark (Calotaxus thornii) granules.

[0042] The invention discloses the construction and application of a thin layer identification method for Chuantong bark (catalpa asiatica) granules and Chuantong bark (catalpa asiatica hairy-leaved) granules. The method has good stability, high precision, good reproducibility, is convenient and easy to master. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0044] Figure 1 For the identification of the basal origin of the Chuantong bark by the developer 1 in the present invention;

[0045] Figure 2 For the identification of the base of Chuantong bark by the developing agent 2 in the present invention;

[0046] Figure 3 For the identification of the base of Chuantong bark by the developing agent 3 in the present invention;

[0047] Figure 4 This is confirmed by the identification method of the Sichuan Paulownia bark of the present invention;

[0048] Figure 5 For the present invention, the amount of sample is investigated;

[0049] Figure 6 For the specificity investigation of the present invention;

[0050] Figure 7 Investigation of different thin layer plates for the present invention - Tianjin Silida;

[0051] Figure 8 For the present invention different thin layer plate investigation - Merck;

[0052] Figure 9 For the different thin layer plates of the present invention, we investigated - Qingdao Yuminyuan;

[0053] Figure 10 This is an investigation of different temperatures of the present invention (4°C);

[0054] Figure 11 This is an investigation of different temperatures of the present invention (30°C);

[0055] Figure 12 For the investigation of different humidity of the present invention (32% rh);

[0056] Figure 13 This is the investigation of different humidity of the present invention (75% rh);

[0057] Figure 14 This is a multiple batch verification of the present invention. DETAILED DESCRIPTION

[0058] In order to further illustrate the present invention, the following detailed description of a thin layer identification method for granules of different raw material formulas of Sichuan tung bark provided by the present invention is given in combination with the examples, but it should not be understood as limiting the scope of protection of the present invention.

[0059] Instruments and reagents

[0060] instrument

[0061] Hot plate, mortar, thin layer imaging system: CAMAG TLC Visualizer, silica gel G thin layer plate (Qingdao Ocean Chemical Plant, batch number: 20180527, Tianjin Silida Technology Co., Ltd., batch number: 191016, Merck, batch number: HX87183353)

[0062] Reagents

[0063] Chloroform, ethanol, n-butanol, hydrochloric acid, ethyl acetate, and glacial acetic acid were all analytical grade, and water was ultrapure water (prepared in the laboratory).

[0064] Drug testing

[0065] Chuantong bark (Cassia truncatum) control material (Sichuan Weikeqi Biotechnology Co., Ltd., batch number: ycwkq22101407); Chuantong bark (Cassia truncatum) control material (Sichuan Weikeqi Biotechnology Co., Ltd., batch number: ycwkq23011102); Chuantong bark (Cassia truncatum) formulated granules 20302015, 20302016, and 20302017; Chuantong bark (Cassia truncatum) formulated granules 2045016, 2045017, and 2045018.

[0066] Example 1 Investigation of the developing agent

[0067] Take 1g each of the powder of the control medicinal material of Catalpa asiatica, the formulated granules of Catalpa asiatica, the control medicinal material of Catalpa asiatica and the formulated granules of Catalpa asiatica respectively, add 20ml of water and 3-5ml of hydrochloric acid, heat under reflux for hydrolysis for 2 hours, cool, and extract twice with saturated n-butanol with water, 15ml each time, combine the n-butanol extracts, evaporate to dryness, dissolve the residue in 5ml of water, and pass it through a neutral alumina column, add 50ml of water for elution, discard the eluate, and elute with 25ml of ethanol, collect the eluate, recover the solvent to dryness, add 1ml of ethanol to the residue and ultrasonicate for 30min, and take the filtrate as the test solution of Catalpa asiatica, the test solution of Catalpa asiatica, the control medicinal material solution and the control medicinal material solution of Catalpa asiatica.

[0068] According to the thin layer chromatography method (General Rules 0502 of the 2020 edition of the Chinese Pharmacopoeia), 3 to 5 μl of each of the above four solutions were taken and spotted on the same silica gel G thin layer plate, respectively, with petroleum ether (30-60°C)-chloroform-ethanol-glacial acetic acid (15:6:1:0.3) as developing agent 1, n-hexane-ethyl acetate-glacial acetic acid (7:3:0.3) as developing agent 2, and n-butanol-glacial acetic acid-water (4:1:5) as developing agent 3. Develop, take out, dry, spray with 10% sulfuric acid ethanol, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material. See the results. Figures 1 to 3 .

[0069] The results showed that when the developing agent petroleum ether (30-60℃)-chloroform-ethanol-glacial acetic acid (15:6:1:0.3) was used, the spots were clear. The hairy-leaved catalpa granules test sample had obvious spots at Rf=0.250, while the thorny-leaved catalpa granules test sample did not have such spots. This method can be used to achieve the qualitative identification of the Chuantong bark (thorny-leaved catalpa) formula granules and the Chuantong bark (hairy-leaved catalpa) formula granules.

[0070] Example 2 Identification Method Confirmation

[0071] Preparation of test solution: Take 1 g of the powder of this product, add 20 ml of water and 3-5 ml of hydrochloric acid, heat under reflux for hydrolysis for 2 hours, cool, and extract twice with 15 ml of n-butanol saturated with water, combine the n-butanol extracts, evaporate to dryness, dissolve the residue in 5 ml of water, pass through a neutral alumina column, add 50 ml of water to elute, discard the eluate, and elute with 25 ml of ethanol, collect the eluate, recover the solvent to dryness, add 1 ml of ethanol to the residue and ultrasonicate for 30 minutes, and take the filtrate as the test solution.

[0072] Preparation of control medicinal material solution: take 2g of control medicinal material, add 50ml of water, decoct for 30 minutes, filter, evaporate the filtrate to dryness, add 20ml of water and 3-5ml of hydrochloric acid to the residue, heat and reflux to hydrolyze for 2 hours, cool, and extract twice with water-saturated n-butanol, 15ml each time, combine the n-butanol extracts, evaporate to dryness, dissolve the residue in 5ml of water, and pass it through a neutral alumina column, add 50ml of water to elute, discard the eluate, and elute with 25ml of ethanol, collect the eluate, recover the solvent to dryness, add 1mL of ethanol to the residue and ultrasonicate for 30min, and take the filtrate as the control medicinal material solution.

[0073] According to the thin layer chromatography method (General Rules 0502 of the 2020 edition of the Chinese Pharmacopoeia), 3 to 5 μl of each of the above two solutions were taken and spotted on the same silica gel G thin layer plate, and petroleum ether (30 to 60 ° C) - chloroform - ethanol - glacial acetic acid (15: 6: 1: 0.3) was used as the developing agent. After development, the plate was taken out, dried, sprayed with 10% sulfuric acid ethanol, heated at 105 ° C until the spots were clearly colored, and inspected under ultraviolet light (365 nm). In the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding position of the chromatogram of the control medicinal material. The results are shown in Figure 4 .

[0074] The results showed that this method can effectively distinguish the different original medicinal materials of Chuantong bark.

[0075] Example 3 Thin layer method establishment

[0076] 3.1 Preparation of test solution

[0077] Take 1g of the powder of this product, add 20ml of water and 3-5ml of hydrochloric acid, heat under reflux and hydrolyze for 2 hours, cool, and extract twice with water-saturated n-butanol, 15ml each time, combine the n-butanol extracts, evaporate to dryness, dissolve the residue in 5ml of water and pass it through a neutral alumina column, add 50ml of water to elute, discard the eluate, and elute with 25ml of ethanol, collect the eluate, recover the solvent to dryness, add 1mL of ethanol to the residue and ultrasonicate for 30min, and take the filtrate as the test solution.

[0078] 3.2 Preparation of control medicinal material solution

[0079] Take 1 g of the control medicinal material, add 20 ml of water and 3-5 ml of hydrochloric acid, heat under reflux and hydrolyze for 2 hours, cool, and extract twice with water-saturated n-butanol, 15 ml each time, combine the n-butanol extracts, evaporate to dryness, dissolve the residue in 5 ml of water, pass through a neutral alumina column, add 50 ml of water to elute, discard the eluate, and elute with 25 ml of ethanol, collect the eluate, recover the solvent to dryness, add 1 mL of ethanol to the residue and ultrasonicate for 30 minutes, and take the filtrate as the control medicinal material solution.

[0080] 3.3 Assay

[0081] According to the thin layer chromatography method (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia), 3-5 μl of each of the above two solutions were applied to the same silica gel G thin layer plate. The plate was developed with petroleum ether (30-60°C)-chloroform-ethanol-glacial acetic acid (15:6:1:0.3) as the developing solvent. The plate was removed, air-dried, sprayed with 10% ethanolic sulfuric acid, and heated at 105°C until the spots were clearly colored. The plate was then examined under ultraviolet light (365 nm). In the chromatogram of the test sample, a fluorescent spot of the same color appeared at the corresponding position in the chromatogram of the reference sample.

[0082] 3.4 Methodological Investigation

[0083] 3.4.1 Inspection of sampling quantity

[0084] Under the experimental conditions proposed above, 1 μl, 3 μl and 5 μl of the control medicinal material solution of Catalpa strychnifolia, the formula granule solution of Catalpa strychnifolia, the control medicinal material solution of Catalpa strychnifolia and the formula granule solution of Catalpa strychnifolia were spotted on the same silica gel G thin layer plate. Figure 5 . Figure 5 In the figure, 1 to 3 are the sample volumes of 1, 3, and 5 μl of the control medicinal material solution of Catalpa fasciata; 4 to 6 are the sample volumes of 1, 3, and 5 μl of the formula granule solution of Catalpa fasciata; 7 to 9 are the sample volumes of 1, 3, and 5 μl of the control medicinal material solution of Catalpa fasciata; 10 to 12 are the sample volumes of 1, 3, and 5 μl of the formula granule solution of Catalpa fasciata.

[0085] Depend on Figure 5 As can be seen, when 3-5 μl of the control herb solution and the test sample solution were spotted, the spots developed clearly and the resolution was good, confirming the recommended spot size of 3-5 μl. Furthermore, at a shift ratio of 0.242, the hairy-leaved catalpa chinensis exhibited blue fluorescent spots, while the schizonepeta chinensis exhibited no spots, which can be used to distinguish between the different origins of the bark of the tung tree. The presence or absence of blue fluorescent spots at a shift ratio of 0.242 can be used as a distinguishing factor.

[0086] 3.4.2 Specificity Investigation

[0087] According to the above test sample preparation method, negative solution, control medicinal material solution of Chinese catalpa chinensis, Chinese catalpa chinensis granule solution, control medicinal material solution of Chinese catalpa chinensis, and Chinese catalpa chinensis granule solution were prepared respectively, and thin layer identification experiment was carried out. The results are shown in Figure 6 , Figure 6 In the table, 1 is the negative solution; 2 is the control medicinal material solution of Catalpa asiatica; 3 is the formula granule solution of Catalpa asiatica; 4 is the control medicinal material solution of Catalpa asiatica; 5 is the formula granule solution of Catalpa asiatica.

[0088] Depend on Figure 6 It can be seen that the negative sample did not interfere with the samples of Catalpa sutchuenensis and Catalpa truncatula, indicating that the method has good specificity. At the ratio shift value of 0.243, Catalpa truncatula showed blue fluorescent spots, while Catalpa sutchuenensis had no spots.

[0089] 3.4.3 Durability assessment

[0090] 3.4.3.1 Comparison of different thin layer plates

[0091] The tailorable thin layer chromatography plates of Tianjin Silida Technology Co., Ltd., the prefabricated silica gel G plates of Merck and Qingdao Yuminyuan Factory Branch were selected and tested according to the proposed test methods. Figures 7-9 , Figures 7-9 Among them, 1. Catalpa asiatica control medicinal material; 2. Catalpa asiatica formula granules; 3. Catalpa asiatica control medicinal material; 4. Catalpa asiatica formula granules.

[0092] The results showed that the method was robust and met the identification requirements. Furthermore, at Rf values ​​(Rf) of 0.226, 0.219, and 0.229, the granules from the bark of the Chinese tung tree (Callopix pubescens) exhibited fluorescent spots, while the granules from the bark of the Chinese tung tree (Callopix pubescens) did not, thus distinguishing the granules from the granules from the bark of the Chinese tung tree (Callopix pubescens).

[0093] 3.4.3.2 Comparison of different temperatures

[0094] Take the spotted thin layer plate and develop it at a low temperature of 4°C and a high temperature of 30°C. Figures 10-11 , Figures 10-11 Among them, 1. Catalpa asiatica control medicinal material; 2. Catalpa asiatica formula granules; 3. Catalpa asiatica control medicinal material; 4. Catalpa asiatica formula granules.

[0095] Depend on Figures 10-11 It can be seen that this method has good adaptability to different temperatures, and the particles of the Tongpi bark (catalpa asiatica) formula show blue fluorescent spots at the ratio shift values ​​of 0.241 and 0.236, while the particles of the Tongpi bark (catalpa asiatica) formula have no spots.

[0096] 3.4.3.3 Comparison of different humidity levels

[0097] Take the spotted thin layer plate and develop it in 32% and 75% humidity environment respectively. Figures 12-13 . Figures 12-13 Among them, 1. Catalpa asiatica control medicinal material; 2. Catalpa asiatica formula granules; 3. Catalpa asiatica control medicinal material; 4. Catalpa asiatica formula granules.

[0098] As can be seen from the figure, this method has good adaptability to different temperatures, and at the ratio shift values ​​of 0.250 and 0.217, the particles of the Chuantong bark (hairy-leaved Catalpa asiatica) formula show blue fluorescent spots, while the particles of the Chuantong bark (hairy-leaved Catalpa asiatica) formula have no spots.

[0099] 3.4.4 Multi-batch verification

[0100] Take the samples of Catalpa oleracea and Catalpa pubescens for thin layer identification and verification. The results are shown in Figure 14 , Figure 14Among them, 1 is the control medicinal material of Catalpa asiatica; 2-4 are the formula granules of Chuantong bark (Calpa asiatica) 20302015, 20302016, 20302017; 5 is the control medicinal material of Catalpa asiatica; 6-8 are the formula granules of Chuantong bark (Calpa asiatica) 2045016, 2045017, 2045018.

[0101] The results showed that the method can accurately distinguish between granules from the bark of the Chinese tung tree (Callopithecus sibiricum) and granules from the bark of the Chinese tung tree (Callopithecus sibiricum). The granules from the bark of the Chinese tung tree (Callopithecus sibiricum) have a distinct blue fluorescent spot with an Rf value of 0.245, which can be used to distinguish the two.

[0102] 3.5 Determination of the identification point Rf of Chuantong bark (catalpa asiatica) formula granules and Chuantong bark (catalpa asiatica) formula granules

[0103] The data of various ratio shift values ​​in the thin layer chromatography methodology investigation are summarized and the results are shown in Table 1. Based on the summary results, it is determined that the identification spot ratio shift value (Rf) of the Chuantong bark formula granules should be within the range of 10%, and the specified value is 0.235.

[0104] Table 1 Summary of methodological investigation ratios

[0105]

[0106]

[0107] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A thin layer identification method for granules of different raw material formulas of Sichuan tung bark, comprising the following steps: A) Add water and hydrochloric acid to the sample to be tested, heat under reflux for hydrolysis, extract with water-saturated n-butanol, and combine the extracts; The extract is evaporated to dryness, dissolved in water, passed through a neutral alumina column, eluted with water and ethanol in sequence, the ethanol eluted eluate is collected, and the solvent is recovered to dryness; The residue is added with methanol and ultrasonicated, and the filtrate is taken to obtain a test solution; the sample to be tested is the granules of Castanopsis arvense or the granules of Castanopsis pubescens; B) taking a control medicinal material and dissolving it with a solvent to obtain a control medicinal material solution; the control medicinal material is a Catalpa strychnifolia control medicinal material or a Catalpa pubescens control medicinal material; C) subjecting the test sample solution and the control medicinal material solution to thin layer chromatography using a silica gel G thin layer plate and a developing solvent of petroleum ether-chloroform-ethanol-glacial acetic acid; the mass ratio of petroleum ether, chloroform, ethanol, and glacial acetic acid being 15:6:1:0.3; D) Check under ultraviolet light. If a blue fluorescent spot appears at an Rf value of 0.235±10% in the chromatogram of the sample, it is Catalpa ovata. If there is no blue fluorescent spot at the Rf value of 0.235±10%, it is Catalpa ovata.

2. The thin layer identification method of different original formula particles of Sichuan tung bark according to claim 1 is characterized in that: The step B) is specifically as follows: The control medicinal material was added with water and hydrochloric acid, heated under reflux for hydrolysis, extracted with water-saturated n-butanol, and the extracts were combined; The extract is evaporated to dryness, dissolved in water, passed through a neutral alumina column, eluted with water and ethanol in sequence, the ethanol eluted eluate is collected, and the solvent is recovered to dryness; The residue was added with methanol and sonicated, and the filtrate was collected to obtain a control medicinal material solution.

3. The thin layer identification method of different original formula particles of Sichuan tung bark according to claim 1 is characterized in that: The thin layer chromatography sample volume is 3-5 μL.

4. The thin layer identification method of different original formula particles of Sichuan tung bark according to claim 1 is characterized in that: Step D) The wavelength of the ultraviolet lamp is 365nm.

5. The thin layer identification method of different original formula particles of Sichuan tung bark according to claim 1 is characterized in that: The temperature examined in step D) is 4-30°C.

6. The thin layer identification method of different original formula particles of Sichuan Paulownia bark according to claim 1 is characterized in that: The humidity checked in step D) is 32-75% rh.

7. The thin layer identification method of different original formula particles of Sichuan tung bark according to claim 1 is characterized in that: The Rf value is 0.217~0.245.

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