Thin layer chromatography identification method for pumpkin pedicle medicinal materials and their preparations

Abstract

The invention provides a thin layer chromatography identification method for pumpkin stalk medicinal materials and preparations thereof. The method comprises the following steps: taking a sample to be tested, decocting with water, filtering, concentrating, extracting with a first solvent, evaporating to dryness, adding water and hydrochloric acid to the residue for extraction by heating, evaporating to dryness, and dissolving the residue in a second solvent to obtain a test solution; the sample to be tested is the pumpkin stalk medicinal material and the preparation thereof; taking a pumpkin stalk reference medicinal material, decocting with water, filtering, concentrating, extracting with a first solvent, evaporating to dryness, adding water and hydrochloric acid to the residue for extraction by heating, evaporating to dryness, and dissolving the residue in a second solvent to obtain a control medicinal material solution; performing thin layer chromatography on the test solution and the control medicinal material solution, using a silica gel G thin layer plate and n-hexane-ether-glacial acetic acid as a developing solvent; spraying with a phosphomolybdic acid ethanol solution, heating until spots are clear, and inspecting under ultraviolet light, so that fluorescent spots of the same color appear at corresponding positions in the chromatogram of the test sample and the chromatogram of the control medicinal material. The present invention uses the observation of whether the fluorescent spots of the same color appear at the corresponding positions of the thin-layer chromatogram and the chromatogram of the control medicinal material as identification points to quickly and effectively identify the pumpkin pedicle medicinal material and its preparation, thereby ensuring the accuracy of clinical medication.

Description

Technical Field

[0001] The invention relates to the technical field of analysis and detection, in particular to a pumpkin pedicle medicinal material and a thin layer identification method for its preparation. Background Art

[0002] Pumpkin pedicle is the dried pedicle of Cucurbita moschata (Duch.ex Lam.) Duch.ex Poir, a plant of the Cucurbitaceae family. It has the effects of tonifying the middle and replenishing qi, nourishing blood and stabilizing the fetus, detoxifying and reducing swelling. It is mainly used clinically for spleen deficiency and weak qi, and fetal movement disorder; it is also used externally to treat furuncles, carbuncles, and burns.

[0003] Pumpkin stalks primarily contain flavonoids, tetraterpenes, sugars, organic acids, and volatile components. The 2018 edition of the "Shanghai Traditional Chinese Medicine Preparation Specifications" and the 2003 edition of the "Guizhou Province Traditional Chinese Medicine and Ethnic Medicinal Materials Quality Standards" use physical and microscopic identification to identify pumpkin stalk medicinal materials and slices, but do not include thin-layer identification, which hinders quality control of the medicinal materials and slices. Because pumpkin stalk granules are made from pumpkin stalk slices through a series of processes, some of which destroy the original physical and microscopic characteristics, physical and microscopic identification methods are not suitable for the identification of pumpkin stalk extracts and preparations.

[0004] Currently, there is no effective thin-layer identification method for the qualitative identification of pumpkin pedicle medicinal materials and preparations. Therefore, it is urgent to establish a method that can identify pumpkin pedicle medicinal materials and preparations to ensure the accurate clinical use of pumpkin pedicles. Summary of the Invention

[0005] In view of this, the technical problem to be solved by the present invention is to provide a thin layer identification method for pumpkin pedicle medicinal materials and their preparations.

[0006] 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.

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

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

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

[0010] This invention aims to establish a thin-layer chromatography method for the identification of pumpkin stalk medicinal materials and formulated granules. By observing whether the fluorescent spots of the same color are located in the same position in the thin-layer chromatography spectrum of the test sample and the control medicinal material, this method can be used to quickly and effectively identify pumpkin stalk medicinal materials and their preparations. This method fills the gap in the qualitative identification of pumpkin stalks, improves the quality standards of pumpkin stalk medicinal materials and their preparations, and ensures drug safety.

[0011] The present invention provides a thin layer identification method for pumpkin pedicle medicinal materials and preparations thereof, comprising:

[0012] A) taking a sample to be tested, boiling it with water, filtering it, concentrating it, extracting it with a first solvent, evaporating it to dryness, extracting the residue by heating it with water and hydrochloric acid, evaporating it to dryness, and dissolving the residue in a second solvent to obtain a test solution; the sample to be tested is pumpkin pedicle medicinal material and its preparation;

[0013] B) taking a pumpkin pedicle reference medicinal material, decocting it with water, filtering, concentrating, extracting it with a first solvent, evaporating it to dryness, extracting the residue by heating with water and hydrochloric acid, evaporating it to dryness, and dissolving the residue in a second solvent to obtain a reference medicinal material solution;

[0014] 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 n-hexane-ether-glacial acetic acid;

[0015] D) Spray with ethanol solution of phosphomolybdic acid, heat until the spots are clear, and examine under ultraviolet light. In the chromatogram of the test sample, fluorescent spots of the same color will appear at the corresponding positions in the chromatogram of the control medicinal material.

[0016] The present invention provides a method for thin layer chromatography identification of pumpkin pedicle medicinal materials and their preparations. First, a sample to be tested is taken and decocted in water. The decocting time of the present invention is 30 minutes.

[0017] The sample to be tested in the present invention is pumpkin stalk medicinal material, pumpkin stalk formula granules or pumpkin stalk decoction pieces.

[0018] The mass volume ratio of the sample to be tested and water in the present invention is 3g:90-110mL;

[0019] In some specific embodiments, the mass volume ratio of the sample to be tested and water is 3 g:100 mL;

[0020] After decoction, the mixture is filtered, concentrated, and extracted with a first solvent. The mixture is evaporated to dryness, and the residue is heated and extracted with water and hydrochloric acid. The mixture is evaporated to dryness, and the residue is dissolved in a second solvent to obtain a control medicinal material solution. The heating extraction is a heating reflux extraction, and the heating reflux extraction time is 1.5 to 2 hours, more preferably 2 hours.

[0021] In a specific embodiment, the method preferably includes: concentrating the filtrate to 20 ml, extracting by shaking with n-butanol 2-3 times, 20 ml each time, combining the n-butanol extracts, and evaporating to dryness. After evaporation to dryness, adding 20 ml of water and 2 ml of hydrochloric acid to the residue, heating under reflux for 2 hours, evaporating to dryness, and dissolving the residue in 1 ml of methanol to prepare the test solution.

[0022] Take pumpkin pedicle reference medicinal material, add water to decoct, filter, concentrate, extract with a first solvent, evaporate to dryness, add water and hydrochloric acid to the residue and heat extract, evaporate to dryness, and dissolve the residue in a second solvent to obtain a reference medicinal material solution.

[0023] The mass volume ratio of the pumpkin pedicle reference medicinal material and water is 3g:90-110mL;

[0024] In some specific embodiments, the mass volume ratio of the pumpkin pedicle control medicinal material and water is 3g:100mL;

[0025] After decoction, the mixture is filtered, concentrated, and extracted with a first solvent. The mixture is evaporated to dryness, and the residue is heated and extracted with water and hydrochloric acid. The mixture is evaporated to dryness, and the residue is dissolved in a second solvent to obtain a control medicinal material solution. The heating extraction is a heating reflux extraction, and the heating reflux extraction time is 1.5 to 2 hours, more preferably 2 hours.

[0026] In a specific embodiment, the method preferably includes: concentrating the filtrate to 20 ml, extracting with n-butanol by shaking 2-3 times, 20 ml each time, combining the n-butanol extracts, and evaporating to dryness. After evaporation to dryness, adding 20 ml of water and 2 ml of hydrochloric acid to the residue, heating under reflux for 2 hours, evaporating to dryness, and dissolving the residue in 1 ml of methanol to obtain a control medicinal material solution.

[0027] The test solution and the control medicinal material solution were subjected to thin layer chromatography.

[0028] Preferably, the test solution and the control medicinal material solution are respectively spotted on the same silica gel G thin layer plate, developed with a developing agent, removed, dried, sprayed with ethanolic phosphomolybdic acid solution, heated until the spots are clear, and inspected under ultraviolet light. The ultraviolet light wavelength is 365nm.

[0029] According to the present invention, the developing agent is n-hexane-ether-glacial acetic acid;

[0030] In some preferred embodiments of the present invention, the volume ratio of n-hexane-diethyl ether-glacial acetic acid is 7:3:1.

[0031] The thin layer plate described herein is a silica gel G thin layer plate. It can be selected from the tailorable thin layer chromatography plates of Tianjin Silida Technology Co., Ltd., prefabricated silica gel G plates from Qingdao Yuminyuan, and Qingdao Ocean Chemical Plant Branch. Results show good durability. In the chromatogram of the test sample, fluorescent spots of the same color appear at corresponding positions in the chromatogram of the control medicinal material.

[0032] The thin layer chromatography sample volume of the present invention is 4 to 10 μL. When 4 to 10 μL of the control medicinal material solution and the test sample solution are sampled, the fluorescent spots on the thin layer chromatography are clearly colored, confirming that the sample volume of the pumpkin pedicle control medicinal material solution and the pumpkin pedicle medicinal material is 4 to 10 μL. In the test sample chromatogram, a fluorescent spot of the same color appears at the corresponding position in the control medicinal material chromatogram.

[0033] The inspection temperature of the present invention is 4° C. to 35° C. The method of the present invention has good adaptability to different temperatures. In the chromatogram of the test sample, fluorescent spots of the same color appear at positions corresponding to those in the chromatogram of the control medicinal material.

[0034] The inspection humidity of the present invention is 32% rh to 75% rh. The method of the present invention has good adaptability to different humidity. In the chromatogram of the test sample, fluorescent spots of the same color appear at positions corresponding to those in the chromatogram of the control medicinal material.

[0035] The present invention provides a method for identifying pumpkin stalk medicinal materials and melon stalk medicinal materials, and the identification is performed by using the method described in the above technical solution.

[0036] The present invention provides a method for identifying pumpkin stalk medicinal materials and wax gourd stalk medicinal materials, and the identification is performed using the method described in the above technical solution.

[0037] The inventors found that the chromatograms of the test samples of melon stalks and wax gourd stalks showed significant differences in spots at corresponding positions compared with the chromatograms of the control medicinal materials of pumpkin stalks. The pumpkin stalk thin layer identification method established by the present invention has strong specificity and can be used to identify pumpkin stalk medicinal materials, extracts, and preparations.

[0038] The invention provides a thin layer chromatography identification method for pumpkin stalk medicinal materials and preparations thereof, comprising the following steps: A) taking a sample to be tested, boiling it with water, filtering, concentrating, extracting it with a first solvent, evaporating to dryness, adding water and hydrochloric acid to the residue for extraction by heating, evaporating to dryness, and dissolving the residue in a second solvent to obtain a test solution; the sample to be tested is the pumpkin stalk medicinal material and preparations thereof; B) taking a pumpkin stalk control medicinal material, boiling it with water, filtering, concentrating, extracting it with a first solvent, filtering, adding water and hydrochloric acid to the residue for extraction by heating, evaporating to dryness, and dissolving the residue in a second solvent to obtain a control medicinal material solution; and 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 n-hexane-ether-glacial acetic acid.

[0039] D) spraying with phosphomolybdic acid ethanol solution, heating until the spots are clear, and inspecting under ultraviolet light. In the chromatogram of the test sample, fluorescent spots of the same color appear at positions corresponding to the chromatogram of the control medicinal material. The thin-layer chromatography identification method for pumpkin pedicle medicinal materials and preparations thereof established by the present invention uses fluorescent spots of the same color as those of the control medicinal material at positions corresponding to the chromatogram of the thin-layer chromatography to quickly and effectively identify pumpkin pedicle medicinal materials and preparations thereof.

[0040] The invention discloses the construction and application of a thin-layer identification method for pumpkin pedicle medicinal materials and preparations thereof. The method has good stability, high precision, good reproducibility, is convenient and easy to master. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 Spot sampling inspection;

[0042] Figure 2 Exclusive inspection;

[0043] Figure 3 Inspection of different thin layer boards-Tianjin Silida;

[0044] Figure 4 Inspection of different thin layer boards-Qingdao Yuminyuan;

[0045] Figure 5 Investigation of different thin layer plates-Qingdao Ocean;

[0046] Figure 6 Different temperatures -4°C;

[0047] Figure 7 Different temperatures -35°C;

[0048] Figure 8 Different humidity - 32%;

[0049] Figure 9 Different humidity - 75%;

[0050] Figure 10 Multiple batches of pumpkin pedicle medicinal materials were verified;

[0051] Figure 11 Multiple batches of pumpkin stem formula granules were verified;

[0052] Figure 12 This is an identification chart of melon stem medicinal materials, wax gourd stem medicinal materials and pumpkin stem medicinal materials;

[0053] Figure 13 Comparative Example 1 thin layer diagram;

[0054] Figure 14 Comparative Example 2 thin layer diagram. DETAILED DESCRIPTION

[0055] To further illustrate the present invention, the following describes in detail a method for constructing a thin-layer identification method for pumpkin pedicle medicinal materials and their preparations provided by the present invention in conjunction with examples.

[0056] Hot plate, mortar, thin layer imaging system: CAMAG TLC Visualizer, silica gel G thin layer plate (Qingdao Ocean Chemical Plant, batch number: 20220307, ​​Tianjin Silida Technology Co., Ltd., batch number: 191026; Qingdao Yuminyuan, batch number: 20200613)

[0057] Chloroform, anhydrous ethanol, n-hexane, ether, glacial acetic acid, methanol, toluene, glacial acetic acid, phosphomolybdic acid, etc. were all analytical grade, and water was ultrapure water.

[0058] Pumpkin stalk control medicinal material (Chengdu Desite Biotechnology Co., Ltd., batch number: DSTYN003401); pumpkin stalk medicinal materials (XXLS202309297, XXLS2023092928, XXLS202309299); pumpkin stalk formula granules (batch numbers: B2311028, B2311029, B23119030); melon stalk medicinal material (XXLS202301026); winter melon stalk medicinal material (XXLS202306023)

[0059] Example 1

[0060] 1.1 Preparation of test solution

[0061] Take 3g of pumpkin stalk medicinal material powder, add 100ml of water, decoct for 30 minutes, filter, concentrate the filtrate to 20ml, add n-butanol and shake to extract 3 times, 20ml each time, combine the n-butanol extracts, evaporate to dryness, add 20ml of water to the residue, add 2ml of hydrochloric acid, heat and reflux to extract for 2 hours, evaporate to dryness, add 1ml of methanol to the residue to dissolve it, and use it as the test solution.

[0062] 1.2 Preparation of control medicinal material solution

[0063] Take 3g of pumpkin stalk reference medicinal material, add 100ml of water, decoct for 30 minutes, filter, concentrate the filtrate to 20ml, add n-butanol and shake to extract 3 times, 20ml each time, combine the n-butanol extracts, evaporate to dryness, add 20ml of water to the residue, add 2ml of hydrochloric acid, heat and reflux to extract for 2 hours, evaporate to dryness, add 1ml of methanol to the residue to dissolve it, and use it as the test solution.

[0064] 1.3 Assay

[0065] According to the thin-layer chromatography method (Chinese Pharmacopoeia 2020 Edition, General Chapter 0502), 10 μl of each of the two solutions was applied to the same silica gel G thin-layer plate. Develop the plate with n-hexane-ether-glacial acetic acid (7:3:1) as the developing solvent. Remove the plate, allow it to air dry, and spray it with 10% ethanolic phosphomolybdic acid solution. Heat it at 105°C until the spots are clearly colored. Examine the plate under ultraviolet light (365 nm). In the chromatogram of the test sample, a fluorescent spot of the same color will appear at the corresponding position in the chromatogram of the control medicinal material.

[0066] Example 2 Methodological Investigation

[0067] 2.4.1 Sampling quantity inspection

[0068] Under the experimental conditions proposed above, 4, 6, 8, and 10 μl of pumpkin pedicle control medicinal material solution and 4, 6, 8, and 10 μl of pumpkin pedicle medicinal material solution were spotted on the same silica gel G thin layer plate. The results were shown in the following table. Figure 1 As can be seen from the figure, when 4-10 μl of the control herbal solution and the test solution are applied, the fluorescent spots on the thin-layer chromatography are clearly colored. The applied volume for the pumpkin pedicle control herbal solution and the pumpkin pedicle is determined to be 4-10 μl. In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions on the chromatogram of the control herbal solution. Figure 1 Sample volume inspection; Note: 1 to 4 are pumpkin pedicle control medicinal materials 4, 6, 8, 10 μl; 5 to 8 are pumpkin pedicle medicinal materials 4, 6, 8, 10 μl.

[0069] 2.4.2 Specificity Investigation

[0070] According to the above test sample preparation method, negative solution, pumpkin pedicle control medicinal material solution, and pumpkin pedicle medicinal material solution were prepared respectively, and spotted on the same thin layer plate. The plate was developed according to the proposed thin layer chromatography conditions and the results were examined. Figure 2 As can be seen from the figure, the negative sample has no interference with the pumpkin pedicle medicinal material test sample. This method has good specificity and can be used to identify pumpkin pedicle medicinal materials. Figure 2 Specificity investigation; Note: 1 is the negative solution; 2 is the pumpkin pedicle control medicinal material; 4 is the pumpkin pedicle medicinal material.

[0071] 2.4.3 Durability Investigation

[0072] 2.4.3.1 Comparison of different thin layer plates

[0073] The tailorable thin layer chromatography plates of Tianjin Silida Technology Co., Ltd., the prefabricated silica gel G plates of Qingdao Yuminyuan and Qingdao Ocean Chemical Plant Branch were selected and tested according to the proposed test methods. Figure 3-5 The results showed that all three brands of TLC plates met the identification requirements, indicating that the method is robust. In the chromatograms of the test samples, fluorescent spots of the same color appeared at the corresponding positions in the chromatograms of the control herbs. Figure 3 Different thin layer plate inspection - Tianjin Silida. Note: 1 is the pumpkin pedicle control medicinal material; 2 is the pumpkin pedicle medicinal material. Figure 4 Investigation of different thin layer plates - Qingdao Yuminyuan; Note: 1 is pumpkin pedicle control medicinal material; 2 is pumpkin pedicle medicinal material. Figure 5 Different thin layer plate inspection - Qingdao Ocean; Note: 1 is pumpkin pedicle control medicinal material; 2 is pumpkin pedicle medicinal material

[0074] 2.4.3.2 Comparison of different temperatures

[0075] Take the spotted thin layer plate and develop it at a low temperature of 4°C and a high temperature of 35°C. Figure 6-7 It can be seen that this method has good durability under different temperatures. In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions in the chromatogram of the control medicinal material. Figure 6 Different temperatures -4 ° C. Note: 1 is the pumpkin pedicle control medicinal material; 2 is the pumpkin pedicle medicinal material; Figure 7 Different temperature -35℃; Note: 1 is pumpkin pedicle control medicinal material; 2 is pumpkin pedicle medicinal material.

[0076] 2.4.3.3 Comparison of different humidity levels

[0077] Take the spotted thin layer plate and develop it in 32% rh and 75% rh humidity environment respectively. Figure 8-9 As can be seen from the figure, this method has good durability under different humidity conditions. In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions in the chromatogram of the control medicinal material. Figure 8 Different humidity -32%; Note: 1 is the pumpkin stalk control medicinal material; 2 is the pumpkin stalk medicinal material. Figure 9 Different humidity -75%; Note: 1 is the pumpkin stalk control medicinal material; 2 is the pumpkin stalk medicinal material.

[0078] 2.4.4 Medicinal Material Verification

[0079] Three batches of pumpkin pedicle medicinal materials were subjected to thin layer identification and verification. The experimental results are shown in Figure 10 The results showed that in the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding positions in the chromatogram of the control medicinal material. Figure 10 Multiple batches of pumpkin stalk medicinal materials were verified; Note: 1 is the pumpkin stalk control medicinal material; 2-4 are pumpkin stalk medicinal materials XXLS202309297, XXLS2023092928, and XXLS202309299.

[0080] 2.4.5 Formula Particle Verification

[0081] Take 2g of this product, grind it into powder, add 20ml of water and ultrasonically dissolve it, add n-butanol and shake to extract 3 times, 20ml each time, combine the n-butanol extracts, evaporate to dryness, add 20ml of water to the residue, add 2ml of hydrochloric acid, heat and reflux to extract for 2 hours, evaporate to dryness, and dissolve the residue in 1ml of methanol as the test solution.

[0082] The results of the thin layer identification and verification experiments on three batches of pumpkin stem formula granules are shown in Figure 11 The results showed that in the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding positions in the chromatogram of the control medicinal material. Figure 11 Multiple batches of pumpkin stalk formula granules were validated; Note: 1 is pumpkin stalk control medicinal material; 2-4 are pumpkin stalk formula granules B2311028, B2311029, and B23119030.

[0083] Example 3 Identification of Melon Pedicle Medicinal Materials, Wax Gourd Pedicle Medicinal Materials and Pumpkin Pedicle Medicinal Materials

[0084] Preparation of test solution

[0085] Take 3g of pumpkin stalk, melon stalk and wax gourd stalk powder respectively, add 100ml of water, boil for 30 minutes, filter, concentrate the filtrate to 20ml, add n-butanol and shake to extract 3 times, 20ml each time, combine the n-butanol extracts, evaporate to dryness, add 20ml of water to the residue, add 2ml of hydrochloric acid, heat and reflux to extract for 2 hours, evaporate to dryness, and dissolve the residue in 1ml of methanol as the test solution.

[0086] 3.2. Preparation of control medicinal material solution

[0087] Take 3g of pumpkin stalk reference medicinal material, add 100ml of water, decoct for 30 minutes, filter, concentrate the filtrate to 20ml, add n-butanol and shake to extract 3 times, 20ml each time, combine the n-butanol extracts, evaporate to dryness, add 20ml of water to the residue, add 2ml of hydrochloric acid, heat and reflux to extract for 2 hours, evaporate to dryness, dissolve the residue in 1ml of methanol, and use it as the test solution.

[0088] 3.3 Assay

[0089] According to the thin layer chromatography method (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia), 10 μl of the test sample solution and the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate, and developed with n-hexane-ether-glacial acetic acid (7:3:1) as the developing agent. The plate was taken out, dried, sprayed with 10% phosphomolybdic acid ethanol solution, heated at 105°C until the spots were clearly colored, and inspected under ultraviolet light (365 nm).

[0090] See attached for the results Figure 12 . Figure 12This is an identification chart for melon stem, wax gourd stem, and pumpkin stem medicinal materials. Note: 1 is a pumpkin stem control medicinal material, 2 is a pumpkin stem medicinal material, 3 is a melon stem medicinal material, and 4 is a wax gourd stem medicinal material. The results show that the chromatograms of the melon stem and wax gourd stem test samples show significant differences in the corresponding spots compared to the pumpkin stem control medicinal material. The established pumpkin stem thin-layer chromatography identification method is highly specific and can be used to identify pumpkin stem medicinal materials, extracts, and preparations.

[0091] Comparative Example 1

[0092] Take 3g of pumpkin stalk powder, add 100ml of water, and boil for 30 minutes. Filter, concentrate the filtrate to 20ml, and extract with n-butanol three times, shaking 20ml each time. Combine the n-butanol extracts, evaporate to dryness, and dissolve the residue in 1ml of methanol to prepare the test solution. Take 3g of pumpkin stalk control medicinal material, add 100ml of water, boil for 30 minutes, filter, and evaporate the filtrate to dryness. Prepare the control medicinal material solution using the same method as above, starting from "extraction with n-butanol and shaking." According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 Edition, General Chapter 0502), 15μl of each solution was spotted on the same silica gel G thin layer plate using n-hexane-ether-glacial acetic acid (7:3:1) as the developing solvent. Develop, remove, air dry, spray with 10% sulfuric acid in ethanol, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365nm). In the chromatogram of the test sample, a fluorescent spot of the same color appears at the corresponding position in the chromatogram of the control medicinal material. Figure 13 . Figure 13 Thin layer diagram of comparative example 1; among them, 1 is the pumpkin pedicle control medicinal material; 2 to 4 are pumpkin pedicle medicinal materials.

[0093] The results showed that using the above-mentioned thin layer identification method, there were no obvious spots between the pumpkin pedicle control medicinal materials and the medicinal materials, and qualitative identification of pumpkin pedicle medicinal materials could not be achieved.

[0094] Comparative Example 2

[0095] Take 2g of pumpkin stalk medicinal material powder, add 20ml of methanol, ultrasonicate for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of methanol to the residue to dissolve it, and use it as the test solution. Take another 2g of pumpkin stalk control medicinal material, and prepare the control medicinal material solution in the same way. According to the thin layer chromatography method (General Rules 0502 of the 2020 Edition of the Chinese Pharmacopoeia), 10μl of each of the above two solutions were taken and spotted on the same silica gel G thin layer plate, and toluene-acetone-glacial acetic acid (8:3:0.3) was used as the developing agent. Develop, take out, dry, spray with 10% phosphomolybdic acid ethanol solution, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365nm). In the test chromatogram, fluorescent spots of the same color appear at the corresponding position of the control medicinal material chromatogram. The experimental results are shown in Figure 14 . Figure 14 Thin layer diagram of comparative example 2; 1-2 are pumpkin pedicle control medicinal materials; 3-4 are pumpkin medicinal materials.

[0096] The results showed that using the above-mentioned thin layer identification method, there were no obvious spots between the pumpkin pedicle control medicinal materials and the medicinal materials, and the qualitative identification of the pumpkin pedicle medicinal materials could not be achieved.

[0097] 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 method for thin-layer identification of pumpkin pedicle medicinal materials and their preparations, comprising: A) A sample to be tested is decocted with water, filtered, concentrated, extracted with a first solvent, evaporated to dryness, the residue is extracted by heating with water and hydrochloric acid, evaporated to dryness, and the residue is dissolved in a second solvent to obtain a test solution; the sample to be tested is pumpkin pedicle medicinal material and its preparation; the first solvent is n-butanol; and the second solvent is methanol; B) taking a pumpkin stalk as a control medicinal material, decocting it with water, filtering, concentrating, extracting it with a first solvent, evaporating it to dryness, extracting the residue by heating with water and hydrochloric acid, evaporating it to dryness, and dissolving the residue in a second solvent to obtain a control medicinal material solution; 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 n-hexane-diethyl ether-glacial acetic acid in a mass ratio of 7:3:1; D) Spray with ethanol solution of phosphomolybdic acid, heat until the spots are clear, and examine under ultraviolet light. In the chromatogram of the test sample, fluorescent spots of the same color will appear at the corresponding positions in the chromatogram of the control medicinal material.

2. The method according to claim 1, characterized in that In step A), the mass volume ratio of the sample to be tested and water is 3 g: 90-110 mL; the decoction time is 30 min; the number of n-butanol extractions is 2-3 times; the heating extraction is heating reflux extraction, and the heating reflux extraction time is 1.5-2 h; The sample to be tested is pumpkin stalk medicinal material, pumpkin stalk formula granules or pumpkin stalk decoction pieces.

3. The method according to claim 1, characterized in that In step B), the first solvent is n-butanol; the second solvent is methanol; the mass volume ratio of the pumpkin pedicle reference medicinal material to water is 3g:90-110mL; the decoction time is 30min; the number of n-butanol extractions is 2-3 times; the heating extraction is heating reflux extraction, and the heating reflux extraction time is 1.5-2h.

4. The method according to claim 1, wherein The thin layer chromatography sample volume is 4-10 μL.

5. The method according to claim 1, wherein Step D) The wavelength of the ultraviolet light is 365 nm; the concentration of the phosphomolybdic acid ethanol solution is 10%; and the heating temperature is 105°C.

6. The method according to claim 1, characterized in that Step D) Check the temperature between 4°C and 35°C.

7. The method according to claim 1, characterized in that Step D) Check the humidity to be between 32% RH and 75% RH.

8. A method for identifying pumpkin stalk medicinal materials and melon stalk medicinal materials, characterized in that: The identification is performed using the method described in any one of claims 1 to 7.

9. A method for identifying pumpkin stalk medicinal materials and wax gourd stalk medicinal materials, characterized in that: The identification is performed using the method described in any one of claims 1 to 7.

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