A thin layer identification method for products of Momordica charantia

Abstract

The invention belongs to the technical field of identification of traditional Chinese medicines, and specifically relates to a thin-layer identification method for products of the Momordica cochinchinensis family. The thin-layer identification method comprises the following steps: when preparing a test solution, the extraction solvent comprises a water-saturated n-butanol solution; a Momordica cochinchinensis control medicinal material is used as a reference; the test solution and the control medicinal material solution are taken, spotted, a mixed solution of ethyl acetate, anhydrous ethanol, water and glacial acetic acid is used as a developing agent, developed, taken out and inspected. The thin-layer identification method has the advantages of simple preparation of the test solution and non-toxicity, etc., the obtained thin-layer chromatogram has rich spot information, good separation, clear color development of the chromatogram, accurate identification results, and is conducive to the quality control of Momordica cochinchinensis products.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine identification, and particularly relates to a thin layer identification method for Momordica charantia products. Background Art

[0002] The 2020 edition of the Chinese Pharmacopoeia stipulates that Momordica charantia is a plant of the Cucurbitaceae family. Momordica cochinchinensis (Lour.) Spreng. In winter, the ripe fruits are harvested, cut open, and dried until half dry. The pulp is removed, the seeds are taken out, and dried.

[0003] The chemical composition of Momordica cochinchinensis products obtained by different processing methods is slightly different. According to the processing method, Momordica cochinchinensis can be divided into Momordica cochinchinensis kernel and Momordica cochinchinensis frost. Momordica cochinchinensis kernel is obtained after the Momordica cochinchinensis is shelled and kernel is taken out. It was first recorded in "Rihuazi" and has the effects of reducing swelling, dispersing knots, detoxifying, chasing away wind and relieving pain. It tastes bitter, slightly sweet, warm in nature, and toxic. Take the clean Momordica cochinchinensis kernel, stir-fry it, grind it into powder, wrap it with paper, and pressurize it to remove oil to obtain Momordica cochinchinensis frost.

[0004] Thin layer chromatography has the characteristics of easy operation, simple equipment, and easy color development, and is of great significance in the qualitative identification of traditional Chinese medicine. The existing technology often uses the following methods to identify products of the Momordica charantia: (1) petroleum ether-chloroform is used as the extraction solvent of the test sample, and chloroform-methanol-water is used as the developing agent; (2) methanol is used as the extraction solvent of the test sample, and formic acid is used as one of the developing agent components. However, the above methods all have defects to varying degrees. Taking method (1) as an example, petroleum ether-chloroform is used as the extraction solvent, the reagent is highly toxic, the sample preparation time is long, the operation is complicated, and the spot information cannot correspond to the same position. The R in method (2) f The spot separation is poor, the spot information is little, and it cannot meet the quality control requirements well. Summary of the invention

[0005] Therefore, the technical problem to be solved by the present invention is to overcome the defects of the prior art in identifying Momordica charantia products, such as long sample preparation time for the test solution, little spot information, poor separation, and failure to meet quality control requirements, thereby providing a thin layer identification method for Momordica charantia products.

[0006] To this end, the present invention provides the following technical solutions.

[0007] The present invention provides a thin layer identification method for Momordica charantia products, comprising the following steps:

[0008] Preparation of the test solution: When preparing the test solution, the extraction solvent includes water and n-butanol;

[0009] Preparation of control medicinal material solution: Momordica charantia control medicinal material was used as the control substance;

[0010] Take the test sample solution and the control medicinal material solution, spot the samples, use a mixture of ethyl acetate, anhydrous ethanol, water and glacial acetic acid as the developing agent, develop, take out and inspect.

[0011] The developing agent includes ethyl acetate, anhydrous ethanol, water and glacial acetic acid in a volume ratio of (7.5-8.5):(1-2):(0.1-1):(1.5-2).

[0012] The developing agent includes ethyl acetate, anhydrous ethanol, water and glacial acetic acid in a volume ratio of 8:1:1:2.

[0013] The Momordica cochinchinensis products are selected from Momordica cochinchinensis medicinal materials, Momordica cochinchinensis decoction pieces, Momordica cochinchinensis frost decoction pieces, Momordica cochinchinensis decoction standard decoction freeze-dried powder or Momordica cochinchinensis formula granules.

[0014] In the thin layer identification method, when preparing the test solution, the extraction solvent includes water-saturated n-butanol;

[0015] Preferably, the method for preparing the test solution comprises: taking the test sample, adding water, ultrasonicating, filtering, extracting the filtrate with water-saturated n-butanol, evaporating to dryness, and adding methanol to the residue to dissolve it.

[0016] The ratio of the mass of the test sample to the volume of water-saturated n-butanol is 0.03-0.13 g:1 ml.

[0017] In the thin layer identification method, when preparing the reference medicinal material solution, the extraction solvent includes water-saturated n-butanol;

[0018] Preferably, the preparation method of the control medicinal material solution comprises: taking a reference substance, adding water, ultrasonicating, filtering, extracting the filtrate with water-saturated n-butanol, evaporating to dryness, and adding methanol to the residue to dissolve it.

[0019] The ratio of the mass of the reference substance to the volume of water-saturated n-butanol is 0.03-0.13 g:1 ml.

[0020] In the thin layer identification method, when performing thin layer identification, the color developer is a 10% sulfuric acid ethanol solution;

[0021] Preferably, before the inspection, the step of heating to 100-110° C. is also included;

[0022] Preferably, the inspection is performed under ultraviolet light.

[0023] In the thin layer identification method, the spotting volume is 5-10 μL.

[0024] The technical solution of the present invention has the following advantages:

[0025] 1. The thin-layer identification method of Momordica cochinchinensis products provided by the present invention comprises the following steps: when preparing the test solution, the extraction solvent comprises a water-saturated n-butanol solution; Momordica cochinchinensis control medicinal material is used as a reference; the test solution and the reference medicinal material solution are taken, spotted, a mixed solution of ethyl acetate, anhydrous ethanol, water and glacial acetic acid is used as a developing agent, developed, taken out and inspected. The thin-layer identification method has the advantages of simple preparation of the test solution and non-toxicity, and the obtained thin-layer chromatogram has rich spot information, good separation, clear color development of the chromatogram, accurate identification results, and is conducive to the quality control of Momordica cochinchinensis products.

[0026] The test solution extraction solvent of the present invention includes water-saturated n-butanol, and the main chemical components in the Momordica cochinchinensis products, such as terpenes, saponins, etc., can be extracted. Compared with the prior art for extracting the hydrolyzed product gypsophila saponin, the overall quality of the Momordica cochinchinensis products can be more accurately reflected, and the quality control is convenient. Ethyl acetate, anhydrous ethanol, water and glacial acetic acid are used as developing agents, and the developing agent system is matched with the test solution obtained by extraction with water-saturated n-butanol, and the obtained thin layer chromatography Rf value is appropriate, the spot information is rich, the separation is good, the color development is clear, and the identification result is accurate.

[0027] The thin layer identification method provided by the present invention has good durability against environmental factors such as temperature and humidity, and has good reproducibility for thin layer plates from different manufacturers and different operators.

[0028] 2. The thin layer chromatography identification method of Momordica charantia products provided by the present invention. The test solution provided by the present invention has the advantages of simple and rapid preparation method, and the extraction solvent used is non-toxic. It can comprehensively and sufficiently extract the chemical components in the Momordica charantia products, providing a basis for examining and obtaining rich spot information of thin layer chromatography, thereby ensuring the quality controllability of Momordica charantia products. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0030] Figure 1 is the thin layer chromatogram obtained in Example 1 of the present invention;

[0031] Figure 2 This is the chromatogram obtained when the sample amount was investigated in Experimental Example 1 of the present invention;

[0032] Figure 3 This is the chromatogram obtained during the durability study in Experimental Example 1 of the present invention;

[0033] Figure 4 This is the chromatogram obtained during the thin layer plate inspection in Experimental Example 1 of the present invention;

[0034] Figure 5 This is the chromatogram obtained during the reproducibility study in Experimental Example 1 of the present invention;

[0035] Figure 6 This is the chromatogram obtained when investigating different batches of Momordica cochinchinensis medicinal materials in Experimental Example 1 of the present invention;

[0036] Figure 7 It is the chromatogram obtained when inspecting different batches of Momordica charantia seeds slices in Experimental Example 1 of the present invention;

[0037] Figure 8 This is a chromatogram obtained when investigating freeze-dried powder of standard decoction of Momordica cochinchinensis seeds in different batches in Experimental Example 1 of the present invention;

[0038] Fig. 9 This is the chromatogram obtained when inspecting different batches of Momordica charantia seed formula granules in Experimental Example 1 of the present invention;

[0039] Fig.10 This is the chromatogram obtained by comparing Experiment 1 with Experiment 2 of the present invention;

[0040] Fig.11 is the chromatogram obtained by comparing Experiment 2 of Experiment 2 of the present invention;

[0041] Fig.12 This is the chromatogram obtained by comparing Experiment 3 with Experiment 2 of the present invention. DETAILED DESCRIPTION

[0042] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0043] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0044] Experimental Materials

[0045] The batch number of the reference medicinal material of Momordica cochinchinensis is: 121418-200501

[0046] The batch number of Momordica cochinchinensis medicinal materials is: K298YC01- K298YC15

[0047] The batch number of Momordica cochinchinensis seed slices is: K298YP01- K298YP15

[0048] The batch number of the freeze-dried powder of Momordica cochinchinensis seed standard decoction is: K298JG01-K298JG15

[0049] The batch number of Momordica cochinchinensis seed granules is: K298CP01- K298CP03

[0050] The batch number of Momordica cochinchinensis frost slices is: K298PZ01

[0051] The batch number of Momordica cochinchinensis shell is: K298YD01

[0052] The batch number of methanol is: 20220214

[0053] The batch number of maltodextrin is: B2102012021011652.

[0054] Example 1

[0055] This embodiment provides a thin layer identification method for products of Momordica charantia, comprising the following steps:

[0056] Preparation of test solution: Take 0.5-2g of the test sample, grind it, add 15ml of water, ultrasonically treat it for 30min, filter it, shake and extract the filtrate with 15ml of water-saturated n-butanol, take water-saturated n-butanol, evaporate it to dryness, add 1ml of methanol to the residue to dissolve it, and obtain the test solution. Among them, when the test sample is a medicinal material, the mass of the test sample is 2g; when the test sample is a decoction piece or granule, the mass of the test sample is 1g; when the test sample is a standard decoction freeze-dried powder, the mass of the test sample is 0.5g.

[0057] Preparation of reference medicinal material solution: Take 2 g of Momordica cochinchinensis reference medicinal material, grind it into powder, add 15 ml of water, ultrasonically treat it for 30 minutes, filter it, shake and extract the filtrate with 15 ml of water-saturated n-butanol, take water-saturated n-butanol, evaporate it to dryness, add 1 ml of methanol to the residue to dissolve it, and obtain a reference medicinal material solution.

[0058] Preparation of negative control solution: Take 2g of maltodextrin, grind it, add 15ml of water, ultrasonically treat it for 30min, filter it, shake and extract the filtrate with 15ml of water-saturated n-butanol, take water-saturated n-butanol, evaporate it to dryness, add 1ml of methanol to the residue to dissolve it, and obtain a negative control solution.

[0059] 5 μL of the test solution, control medicinal material solution and negative control solution were respectively spotted on the same silica gel H thin layer plate (produced by Yantai Institute of Chemical Industry), and ethyl acetate, anhydrous ethanol, water and glacial acetic acid in a volume ratio of 8:1:1:2 were used as developing agents. The plate was developed, taken out, dried, sprayed with 10% ethanolic sulfuric acid solution, heated at 105°C until the spots were clearly colored, and examined under ultraviolet light (365nm).

[0060] Among them, Momordica cochinchinensis medicinal materials, Momordica cochinchinensis decoction pieces, Momordica cochinchinensis decoction standard decoction freeze-dried powder, Momordica cochinchinensis formula granules, Momordica cochinchinensis frost decoction pieces, and Momordica cochinchinensis shell were used as test samples to prepare test sample solutions, and thin layer chromatograms of each test sample were obtained. The results are shown in Figure 1 . The solvent methanol was used as the blank control group. Among them, 1 is the Momordica cochinchinensis control medicinal material solution; 2 is the Momordica cochinchinensis medicinal material; 3 is the Momordica cochinchinensis kernel slices; 4 is the Momordica cochinchinensis kernel standard decoction freeze-dried powder; 5 is the Momordica cochinchinensis kernel formula granules; 6 is the Momordica cochinchinensis frost slices; 7 is the Momordica cochinchinensis shell; 8 is the methanol blank control group; 9 is the negative control solution. The batch numbers of each test product are shown in Table 1:

[0061] Table 1 Batch number of each test product

[0062]

[0063] from Figure 1 It can be seen that methanol solvent has no effect on the TLC spots. The TLCs obtained from different test sample solutions show fluorescent spots of the same color at the corresponding positions of the TLCs of the control medicinal materials, and the identification results are accurate; in addition, the TLCs obtained from different test samples have clear color, rich spot information, and good separation, which is beneficial to the quality control of Momordica cochinchinensis products.

[0064] Experimental Example 1

[0065] (1) Investigation of the amount of sample

[0066] Three batches of freeze-dried powder of standard decoction of Momordica cochinchinensis seeds were taken, with batch numbers K298JG01, K298JG02, and K298JG03, respectively, and the spotting amount was investigated, with the spotting amount being 3 μL, 5 μL, and 10 μL; the test solution was prepared according to the "Preparation of the test solution" in Example 1, and the spotting amount was used as a variable. The thin layer chromatograms of each test sample at different spotting amounts were obtained according to the thin layer identification method in Example 1, and the results are shown in FIG. Figure 2 The batch number, sample volume and corresponding number of the TLC of the freeze-dried powder of the standard decoction of Momordica cochinchinensis seeds are shown in Table 2.

[0067] Table 2 Sample information of each batch of freeze-dried powder of standard decoction of Momordica cochinchinensis seeds

[0068]

[0069] from Figure 2 It can be seen that the optimal spot volume is 5-10 μL, the spots in the thin layer chromatogram are clear, the separation is good, and R f Moderate, best color rendering effect.

[0070] (2) Durability test: the impact of different temperature and humidity conditions on identification results

[0071] Information on Thin Layer Chromatography Conditions: Contact Spotting

[0072] Expanding cylinder specifications: double tank

[0073] Unfolding method and extension distance: upward unfolding, extension distance is 8cm

[0074] Saturation method: Pre-saturation 10min

[0075] Thin layer plate: Silica gel H thin layer plate from Yantai Institute of Chemical Industry;

[0076] Three batches of freeze-dried powder of standard decoction of Momordica cochinchinensis seeds were taken, with batch numbers K298JG01, K298JG02, and K298JG03, respectively. The temperature and humidity were investigated. The temperature of environment ① was 21°C and the humidity was 28%; the temperature of environment ② was 4°C and the humidity was 78%; the temperature of environment ③ was 20°C and the humidity was 23%; the temperature of environment ④ was 21°C and the humidity was 78%; the test solution was prepared according to the "Preparation of Test Solution" in Example 1, and temperature and humidity were used as variables. The thin layer chromatograms of each test sample under different environments were obtained according to the thin layer identification method in Example 1. The results are shown in FIG. Figure 3 .in, Figure 3 a in a is the thin layer chromatogram obtained in environment ①, 1 in a is the thin layer chromatogram of the Momordica cochinchinensis reference medicinal material, 2 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG01; 3 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG02; 4 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG03.

[0077] Figure 3 b in the figure is the thin layer chromatogram obtained in environment ②, 1 in b is the thin layer chromatogram of the Momordica cochinchinensis reference medicinal material, 2 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG01; 3 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG02; 4 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG03.

[0078] Figure 3 c in the figure is the thin layer chromatogram obtained in environment ③. Figure 3In c, 1 is the thin layer chromatogram of the Momordica cochinchinensis reference medicinal material, 2 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG01; 3 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG02; 4 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG03.

[0079] Figure 3 The d in the figure is the thin layer chromatogram obtained in environment ③. Figure 3 In d, 1 is the thin layer chromatogram of the Momordica cochinchinensis reference medicinal material, 2 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG01; 3 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG02; 4 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG03.

[0080] from Figure 3 It can be seen from ad in the figure that under different temperature and humidity conditions, the spots in the chromatograms obtained from the test solution and the control medicinal material solution are clear, and the correspondence and separation effects are good, indicating that the thin layer identification method provided by the present invention has good durability against temperature and humidity.

[0081] (3) Investigation of thin-layer boards produced by different manufacturers

[0082] Three batches of freeze-dried powder of standard decoction of Momordica cochinchinensis seeds, with batch numbers K298JG01, K298JG02 and K298JG03, were taken, and silica gel H thin layer plates produced by different manufacturers were investigated; the test solution was prepared according to the "Preparation of the test solution" in Example 1, and the silica gel H thin layer plate was used as a variable. The thin layer chromatograms of each test sample were obtained according to the thin layer identification method in Example 1, and the results are shown in FIG. Figure 4 .

[0083] Among them, the manufacturers of silica gel H thin layer plates are Tianjin Silida, Yantai Yinlong, Dingkang and Qingdao Ocean. Figure 4 There are 4 groups of test results, which correspond to Tianjin Silida, Yantai Yinlong, Dingkang and Qingdao Ocean from left to right. Among the 4 groups of test results, 1 is the thin layer chromatogram of the Momordica cochinchinensis reference medicinal material (batch number 121418-200501), 2 is the thin layer chromatogram of the freeze-dried powder of the standard decoction of Momordica cochinchinensis kernel with batch number K298JG01; 3 is the thin layer chromatogram of the freeze-dried powder of the standard decoction of Momordica cochinchinensis kernel with batch number K298JG02; 4 is the thin layer chromatogram of the freeze-dried powder of the standard decoction of Momordica cochinchinensis kernel with batch number K298JG03.

[0084] from Figure 4 It can be seen that the silica gel H thin layer plates produced by different manufacturers can achieve good separation effect for the freeze-dried powder of standard decoction of Momordica cochinchinensis seeds, indicating that the thin layer identification method provided by the present invention has good durability for thin layer plates produced by different manufacturers.

[0085] (4) Investigation of reproducibility

[0086] Three batches of freeze-dried powder of standard decoction of Momordica cochinchinensis seeds were taken, with batch numbers K298JG01, K298JG02 and K298JG03, respectively. Different experimenters prepared the test solution according to the "Preparation of the test solution" in Example 1, and obtained the thin layer chromatograms of each test sample according to the thin layer identification method in Example 1. The results are shown in Figure 5 . Figure 5 1 is the thin layer chromatogram of the Momordica cochinchinensis reference medicinal material (batch number 121418-200501), 2 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG01; 3 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG02; 4 is the thin layer chromatogram of the Momordica cochinchinensis standard decoction freeze-dried powder of batch number K298JG03.

[0087] (5) Inspection of different batches of test products

[0088] A. The test sample is Momordica cochinchinensis

[0089] Take 15 batches of Momordica cochinchinensis medicinal materials, with batch numbers K298YC01, K298YC02, K298YC03, K298YC04, K298YC05, K298YC06, K298YC07, K298YC08, K298YC09, K298YC10, K298YC11, K298YC12, K298YC13, K298YC14 and K298YC15, prepare test solution according to the "Preparation of test solution" in Example 1, and obtain thin layer chromatograms of each test sample according to the thin layer identification method in Example 1. The results are shown in FIG. Figure 6 .in, Figure 6 1 is the Momordica cochinchinensis control medicinal material, and 2-16 correspond to the Momordica cochinchinensis medicinal materials of batch numbers K298YC01-K298YC15 respectively.

[0090] B. The test sample is Momordica cochinchinensis seed slices

[0091] Take 15 batches of Momordica cochinchinensis seeds, batch numbers are K298YP01, K298YP02, K298YP03, K298YP04, K298YP05, K298YP06, K298YP07, K298YP08, K298YP09, K298YP10, K298YP11, K298YP12, K298YP13, K298YP14, K298YP15, prepare test solution according to the "Preparation of test solution" in Example 1, and obtain thin layer chromatograms of each test sample according to the thin layer identification method in Example 1. The results are shown in FIG. Figure 7 .in, Figure 7 1 is the Momordica cochinchinensis reference medicinal material, and 2-16 correspond to the Momordica cochinchinensis kernel decoction pieces of batch numbers K298YP01-K298YP 15, respectively.

[0092] C. The test sample is freeze-dried powder of standard decoction of Momordica cochinchinensis seeds

[0093] Take 15 batches of freeze-dried powder of standard decoction of Momordica cochinchinensis seeds, with batch numbers K298JG01, K298JG02, K298JG03, K298JG04, K298JG05, K298JG06, K298JG07, K298JG08, K298JG09, K298JG10, K298JG11, K298JG12, K298JG13, K298JG14, and K298JG15, respectively, and prepare test solutions according to the "Preparation of Test Solution" in Example 1. According to the thin layer identification method in Example 1, thin layer chromatograms of each test sample are obtained respectively. The results are shown in FIG. Figure 8 .in, Figure 8 1 is the Momordica cochinchinensis reference medicinal material, and 2-16 correspond to the Momordica cochinchinensis kernel standard decoction freeze-dried powder of batch numbers K298JG01-K298JG15, respectively.

[0094] D. The test sample is Momordica cochinchinensis seed granules

[0095] Three batches of Momordica cochinchinensis seed granules were taken, with batch numbers K298CP01, K298CP02, and K298CP03, respectively. Test solutions were prepared according to the "Preparation of Test Solution" in Example 1. Thin layer chromatograms of each test sample were obtained according to the thin layer identification method in Example 1. The results are shown in Fig. 9 .in, Fig. 9 1 is the Momordica cochinchinensis control medicinal material, and 2-4 correspond to the Momordica cochinchinensis kernel formula granules of batch numbers K298CP01-K298CP03 respectively.

[0096] As can be seen from the figure, the thin layer identification method of the present invention can be applied to Momordica cochinchinensis medicinal materials, Momordica cochinchinensis kernel decoction pieces, Momordica cochinchinensis kernel standard decoction freeze-dried powder, Momordica cochinchinensis kernel formula granules and Momordica cochinchinensis cream.

[0097] Experimental Example 2

[0098] This experimental example provides a comparative experiment, as follows:

[0099] (1) Comparative experiment 1 Investigation of the extraction solvent of the test solution

[0100] When preparing the test solution, methanol is used as the extraction solvent. The preparation method of the test solution includes: taking 2g of Momordica cochinchinensis medicinal material test sample, adding 30ml of methanol, soaking for 1h, ultrasonic treatment for 15min, filtering, accurately aspirating 20ml of the filtrate, water bathing to dryness, adding methanol to make the volume to 1ml, and obtaining the test solution. According to the thin layer identification method of Example 1, the thin layer chromatogram of comparative experiment 1 is obtained, as shown in FIG. Fig.10 .

[0101] Fig.10 3 is the thin layer chromatogram obtained by the thin layer identification method in Example 1 for the test sample of this comparative experiment 1, 4 is the control medicinal material solution prepared from the Momordica charantia reference medicinal material according to the "control medicinal material solution" in Example 1, and then the chromatogram is obtained by the thin layer identification method of Example 1, and 5 is the test sample solution prepared from the Momordica charantia kernel decoction piece according to the "test sample solution" in Example 1, and then the chromatogram is obtained by the thin layer identification method of Example 1.

[0102] from Fig.10 It can be seen that the test solution obtained by methanol extraction failed to achieve the identification effect under the existing conditions, and the chromatographic information was aggregated and could not be separated. It is speculated that this is related to the complexity of the extracted component information. Therefore, methanol is not suitable as the extraction solvent for this identification method.

[0103] (2) Comparative experiment 2: Investigation of developing agent

[0104] Taking the developing agent as a variable, the test sample solution was prepared according to the "Preparation of the Test Sample Solution" in Example 1, and the thin layer chromatograms of each test sample under different developing agents were obtained according to the thin layer identification method in Example 1. The results are shown in Fig.11 .

[0105] Developing agent A: ethyl acetate, ethanol and formic acid in a volume ratio of 6:1:0.1

[0106] Developing solvent B: ethyl acetate, ethanol and glacial acetic acid in a volume ratio of 4:1:1

[0107] Developing solvent C: ethyl acetate, anhydrous ethanol, water and formic acid in a volume ratio of 8:1:1:2

[0108] The thin layer chromatogram obtained with developing agent A is shown in Fig.11 Left: The thin layer chromatogram obtained with developing agent B is shown in Fig.11 Middle figure, the thin layer chromatogram obtained by developing agent C is shown in Fig.11Right figure; Among them, 3 in each figure is a test solution prepared from Momordica cochinchinensis medicinal material according to the "test solution" in comparative experiment 1, using the developing agents AC of this comparative experiment as developing agents, respectively, and adopting the thin layer identification method of Example 1 to obtain a chromatogram; 4 is a control medicinal material prepared from Momordica cochinchinensis according to the "control medicinal material solution" in Example 1, and then using the developing agents AC of this comparative experiment as developing agents, respectively, and obtaining a chromatogram according to the method in Example 1; 5 is a test solution prepared from Momordica cochinchinensis kernel decoction piece according to the "test solution" in Example 1, and then using the developing agents AC of this comparative experiment as developing agents, respectively, and obtaining a chromatogram according to the method in Example 1.

[0109] from Fig.11 It can be seen that after adjusting the developing agent, only two spots are colored in the thin layer chromatogram obtained by developing agent A, and the clarity is not high. The thin layer chromatogram R obtained by developing agent B f The value decreases, and the separation of the three spots at the bottom is poor. The spots of the thin layer chromatogram obtained by developing agent C move upward as a whole, and the spots cannot be completely separated, and the identification effect cannot be achieved.

[0110] (3) Comparative study of the identification method in Experiment 3

[0111] Preparation of test solution: Take 2 g of Momordica charantia test sample, add 30 ml of methanol, soak for 1 hour, ultrasonically treat for 15 min, filter, accurately pipette 20 ml of the filtrate, dry it in a water bath, add methanol to make up to 1 ml, and the test solution is obtained.

[0112] Ethyl acetate, ethanol and formic acid in a volume ratio of 6:1:0.1 were used as the developing agent, the samples were taken out and dried, and 5% vanillin ethanol-10% sulfuric acid ethanol solution in a volume ratio of 1:20 was used as the color developer and the samples were examined under daylight conditions. Fig.12 .

[0113] Among them, 3 is the chromatogram obtained in this comparative experiment; 4 is a chromatogram obtained by using the Momordica charantia control medicinal material to prepare a control medicinal material solution according to the "control medicinal material solution" in Example 1, and then using the comparative experimental identification method; 5 is a chromatogram obtained by using the Momordica charantia kernel decoction piece to prepare a test solution according to the "test solution" in Example 1, and using the comparative experimental identification method.

[0114] from Fig.12 It can be seen that under the conditions of this comparative experiment, the test sample obtained in the comparative experiment has no spot coloring, and a color band accumulation appears at the front position; the test sample and the control medicinal material obtained in the embodiment have only one spot coloring, less information, and unclear coloring.

[0115] By comparing with comparative experiments 1-3, it is shown that the thin layer chromatogram obtained by the thin layer identification method provided by the present invention has rich spot information, good separation, clear color development of the chromatogram, accurate identification results, which is beneficial to the quality control of Momordica charantia products.

[0116] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the scope of protection of the invention.

Claims

1. A thin layer identification method for products of Momordica charantia, characterized in that: The following steps are involved: Preparation of test solution: Take the test sample, add water, sonicate, filter, extract the filtrate with water-saturated n-butanol, evaporate to dryness, and add methanol to the residue to dissolve it; Preparation of control medicinal material solution: Momordica charantia control medicinal material was used as the control substance; Take the test sample solution and the control medicinal material solution, spot them on the silica gel H thin layer plate, use the mixed solution of ethyl acetate, anhydrous ethanol, water and glacial acetic acid with a volume ratio of 8:1:1:2 as the developing agent, develop, take out and inspect; The Momordica cochinchinensis products are selected from Momordica cochinchinensis medicinal materials, Momordica cochinchinensis kernel decoction pieces, Momordica cochinchinensis frost decoction pieces, Momordica cochinchinensis kernel standard decoction freeze-dried powder or Momordica cochinchinensis kernel formula granules.

2. The thin layer identification method according to claim 1, characterized in that: The ratio of the mass of the test sample to the volume of water-saturated n-butanol is 0.03-0.13 g:1 ml.

3. The thin layer identification method according to claim 1, characterized in that: When preparing the control medicinal material solution, the extraction solvent includes water-saturated n-butanol.

4. The thin layer identification method according to claim 3, characterized in that: The preparation method of the control medicinal material solution comprises: taking a control substance, adding water, ultrasonicating, filtering, extracting the filtrate with water-saturated n-butanol, evaporating to dryness, and adding methanol to the residue to dissolve it.

5. The thin layer identification method according to claim 3, characterized in that: The ratio of the mass of the reference substance to the volume of water-saturated n-butanol is 0.03-0.13 g:1 ml.

6. The thin layer identification method according to any one of claims 1 to 5, characterized in that: When performing thin layer identification, the color developer is 10% sulfuric acid ethanol solution.

7. The thin layer identification method according to claim 6, characterized in that: Before the inspection, the method further includes heating to 100-110°C.

8. The thin layer identification method according to claim 7, characterized in that: The inspection was performed under ultraviolet light.

9. The thin layer identification method according to any one of claims 1 to 5, 7 or 8, characterized in that: The sample volume is 5-10 μL.

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