A thin layer identification method for flax seeds

Abstract

The present invention provides a thin-layer chromatography (TLC) identification method for flaxseed, comprising the following steps: S1) extracting and dissolving a test flaxseed to obtain a test solution, wherein the test flaxseed is a flaxseed medicinal material or a flaxseed formula granule; S2) extracting and dissolving a flaxseed control medicinal material to obtain a control medicinal material solution; S3) subjecting the test solution and the control medicinal material solution to thin-layer chromatography, using a silica gel G TLC plate and a developing solvent of petroleum ether, ethyl acetate, and glacial acetic acid; and S4) inspecting under ultraviolet light. In the TLC identification method for flaxseed, by observing whether the test sample and the control flaxseed medicinal material have the same fluorescent spot of the same color at the same position in the TLC, it can be confirmed that the test sample is a flaxseed medicinal material or a flaxseed formula granule. The method can be used to quickly and effectively identify flaxseed medicinal materials or flaxseed formula granules. Experiments have shown that the identification method provided by the present application has good stability, high accuracy, good reproducibility, convenience, and ease of use.

Description

Technical Field

[0001] The invention relates to the technical field of analysis and detection, in particular to a thin layer identification method for flaxseed. Background Art

[0002] Flaxseed is the dried mature seed of Linum usitatissimum L., a plant of the Linaceae family. It contains gum and oil, and has the effects of moisturizing and relieving constipation, nourishing blood and dispelling wind. It is mainly used clinically for intestinal dryness, constipation, dry skin, itching and hair loss.

[0003] The main chemical components of flaxseed are compounds such as cyclic peptides, lignans, flavonoids, and fatty acids. Current flaxseed identification methods focus primarily on character and component identification. For example, Zhou Jingchun conducted a comparative study of flaxseed and chrysanthemum seeds by identifying their origin and place of production, their properties, and their components. While this method is simple, the subjective descriptions passed down orally combined with sensory evaluation through visual observation are easily influenced by individual differences and environmental factors, making objectivity and accuracy difficult to guarantee. Few studies have been reported on the physical and chemical identification of flaxseed, necessitating the development of a thin-layer chromatography (TLC) method for its identification. Summary of the Invention

[0004] The technical problem solved by the present invention is to provide a thin layer identification method for flax seeds. The thin layer identification method provided by the present application can quickly and effectively identify flax seeds as medicinal materials or flax seeds as formula particles.

[0005] In view of this, the present application provides a thin layer identification method for flax seeds, comprising the following steps:

[0006] S1) extracting and dissolving flaxseed to be tested to obtain a test solution, wherein the flaxseed to be tested is flaxseed medicinal material or flaxseed formula granules;

[0007] S2) extracting and dissolving the flaxseed control medicinal material to obtain a control medicinal material solution;

[0008] S3) The test solution and the control medicinal material solution were subjected to thin layer chromatography (TLC); the TLC plate was a silica gel G TLC plate and the developing solvent was petroleum ether-ethyl acetate-glacial acetic acid;

[0009] S4) Place the sample under ultraviolet light for inspection. If a fluorescent spot of the same color appears in the chromatogram of the test sample at the corresponding position as the control medicinal material, the sample is the flaxseed medicinal material or the flaxseed formula granules.

[0010] Preferably, step S1) is specifically:

[0011] When the flaxseed to be tested is linseed medicinal material, the linseed is decocted with water and then filtered, the filtrate is concentrated and then hydrochloric acid is added, and the mixture is heated under reflux for extraction, and the residue is evaporated to dryness and dissolved in water, and then extracted with ethyl acetate, and the residue is evaporated to dryness and dissolved in methanol to obtain a test solution;

[0012] When the flaxseed to be tested is a flaxseed formula granule, the flaxseed formula granule is ground, and then water and hydrochloric acid are added, and reflux extraction is performed. After evaporation to dryness, water is added to the residue, and then extraction is performed with ethyl acetate. After evaporation to dryness again, methanol is added to dissolve the residue to obtain a test solution;

[0013] The ratio of the linseed, the hydrochloric acid and the ethyl acetate is 1 g: (1-3) ml: (2-8) ml.

[0014] Preferably, step S2) is specifically as follows:

[0015] The linseed control medicinal material was decocted with water and filtered. The filtrate was concentrated and then added with hydrochloric acid. The mixture was heated under reflux for extraction. After evaporation, the residue was dissolved in water and extracted with ethyl acetate. The residue was evaporated and dissolved in methanol to obtain the test solution.

[0016] The ratio of the linseed reference medicinal material, the hydrochloric acid and the ethyl acetate is 1 g: (1-3) ml: (2-8) ml.

[0017] Preferably, in step S3), the sample volume for thin layer chromatography detection is 5 to 15 μL.

[0018] Preferably, the volume ratio of the petroleum ether, the ethyl acetate and the glacial acetic acid is (9-11):1:(0.08-0.12).

[0019] Preferably, the volume ratio of the petroleum ether, the ethyl acetate and the glacial acetic acid is 10:1:0.1.

[0020] Preferably, the wavelength of the ultraviolet lamp is 365nm.

[0021] Preferably, the review further includes:

[0022] The developed thin layer plate was sprayed with 10% sulfuric acid ethanol solution and then heated at 100-110°C.

[0023] Preferably, the inspection temperature is 4°C to 35°C.

[0024] Preferably, the humidity under inspection is 32% rh to 75% rh.

[0025] The present application provides a thin-layer chromatography (TLC) identification method for flaxseed. The method first prepares a test solution and a control medicinal material solution, then subjects the test solution and the control medicinal material solution to TLC. If, in the chromatogram of the test sample, a fluorescent spot of the same color appears at a position corresponding to that of the control medicinal material, the sample is identified as flaxseed medicinal material or flaxseed formulated granules. In the TLC identification method provided herein, by observing whether the TLC of the test sample and the control flaxseed medicinal material show fluorescent spots of the same color at the same position, the sample is confirmed to be flaxseed medicinal material or flaxseed formulated granules. The method can be used to quickly and effectively identify flaxseed medicinal materials or flaxseed formulated granules. Experiments have shown that the identification method provided herein is stable, precise, reproducible, convenient, and easy to master. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a thin layer chromatogram of flaxseed as a control medicinal material and flaxseed as a medicinal material with different sample amounts in Example 1 of the present invention;

[0027] Figure 2 The thin layer chromatograms of the negative solution, linseed control medicinal material, and linseed medicinal material in Example 1 of the present invention are shown;

[0028] Figure 3 This is a thin layer chromatogram of the thin layer chromatography plate of Tianjin Silida in Example 1 of the present invention;

[0029] Figure 4 This is a thin layer chromatogram under the Qingdao Yuminyuan thin layer chromatography plate in Example 1 of the present invention;

[0030] Figure 5 This is a thin layer chromatogram of the Qingdao Ocean thin layer chromatography plate in Example 1 of the present invention;

[0031] Figure 6 This is a thin layer chromatogram of the 4°C flaxseed control medicinal material and the flaxseed medicinal material of Example 1 of the present invention;

[0032] Figure 7 This is a thin layer chromatogram of the 35°C linseed control medicinal material and the linseed medicinal material of Example 1 of the present invention;

[0033] Figure 8 The thin layer chromatograms of the 32% rh linseed control medicinal material and the linseed medicinal material of Example 1 of the present invention are shown;

[0034] Figure 9 The thin layer chromatograms of the 75% rh linseed control medicinal material and the linseed medicinal material of Example 1 of the present invention are shown;

[0035] Figure 10 The thin layer chromatograms of different batches of linseed medicinal materials and linseed control medicinal materials in Example 1 of the present invention are shown;

[0036] Figure 11 The thin layer chromatograms of different batches of flaxseed formula granules and flaxseed control medicinal materials in Example 2 of the present invention are shown;

[0037] Figure 12 This is a thin layer chromatogram of the linseed control medicinal material and the linseed medicinal material in Comparative Example 1 of the present invention;

[0038] Figure 13 This is a thin layer chromatogram of the linseed control medicinal material and the linseed medicinal material in Comparative Example 2 of the present invention. DETAILED DESCRIPTION

[0039] In order to further understand the present invention, preferred embodiments of the present invention are described below in conjunction with examples. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, rather than limiting the claims of the present invention.

[0040] Given the lack of existing physical and chemical identification methods for flaxseed, this application provides a thin-layer chromatography (TLC) identification method for flaxseed. This TLC identification method is used to identify flaxseed medicinal materials and flaxseed formula granules, respectively. By observing whether the thin-layer chromatography spectrum of the test sample contains fluorescent spots of the same color at the same location as the control medicinal material, the method can be used to quickly and effectively identify the flaxseed medicinal material or flaxseed formula granules. This method fills the gap in the qualitative identification of flaxseed, improves the quality standards of flaxseed medicinal materials or flaxseed formula granules, and ensures drug safety. Specifically, the embodiments of the present invention disclose a TLC identification method for flaxseed, comprising the following steps:

[0041] S1) extracting and dissolving flaxseed to be tested to obtain a test solution, wherein the flaxseed to be tested is flaxseed medicinal material or flaxseed formula granules;

[0042] S2) extracting and dissolving the flaxseed control medicinal material to obtain a control medicinal material solution;

[0043] S3) The test solution and the control medicinal material solution were subjected to thin layer chromatography (TLC); the TLC plate was a silica gel G TLC plate and the developing solvent was petroleum ether-ethyl acetate-glacial acetic acid;

[0044] S4) Place the sample under ultraviolet light for inspection. If a fluorescent spot of the same color appears in the chromatogram of the test sample at the corresponding position as the control medicinal material, the sample is the flaxseed medicinal material or the flaxseed formula granules.

[0045] In the flaxseed identification method provided in the present application, in step S1), a test solution is prepared, i.e., the flaxseed to be tested is extracted and then dissolved. In the present application, the flaxseed to be tested is flaxseed medicinal material or flaxseed formula granules. The flaxseed to be tested is extracted with hydrochloric acid and then ethyl acetate, and then dissolved in methanol. When the flaxseed to be tested is flaxseed medicinal material, step S1) is specifically as follows:

[0046] Add linseed to water and boil, then filter. Concentrate the filtrate, add hydrochloric acid, heat under reflux for extraction, evaporate to dryness, dissolve the residue in water, extract with ethyl acetate, evaporate to dryness, dissolve the residue in methanol to obtain the test solution.

[0047] The ratio of the linseed, the hydrochloric acid, and the ethyl acetate is 1 g: (1-3) ml: (2-8) ml; more specifically, the ratio of the linseed, the hydrochloric acid, and the ethyl acetate is 1 g: 1 ml: 5 ml.

[0048] When the flaxseed to be tested is flaxseed formula granules, step S1) is specifically as follows:

[0049] Grind the flaxseed formula granules, add water and hydrochloric acid, reflux extract, evaporate to dryness, add water to the residue, extract with ethyl acetate, evaporate to dryness again, and dissolve the residue in methanol to obtain the test solution;

[0050] In the above process, the ratio of the flaxseed formula particles, the hydrochloric acid, and the ethyl acetate is 1 g: (1-3) ml: (2-8) ml; more specifically, the ratio of the flaxseed formula particles, the hydrochloric acid, and the ethyl acetate is 1 g: 1 ml: 5 ml.

[0051] In step S2), a control medicinal material solution is prepared, that is, the solvent after the linseed control medicinal material is extracted; the preparation method of the control medicinal material solution is the same as the preparation method of the linseed medicinal material solution, which will not be described here in detail.

[0052] After the test solution and control herbal solution are prepared, they are subjected to thin-layer chromatography. Specifically, the test solution and control herbal solution are each spotted onto a single silica gel G thin-layer plate, developed with a developing agent, removed, dried, and examined under ultraviolet light with a wavelength of 365 nm.

[0053] In the above process, the developing agent is petroleum ether, ethyl acetate and glacial acetic acid, and the volume ratio thereof is (9-11):1:(0.08-0.12). In a specific embodiment, the volume ratio of the petroleum ether, ethyl acetate and glacial acetic acid is 10:1:0.1; wherein the boiling point of petroleum ether is 60-90°C.

[0054] The silica gel G thin layer chromatography plates described in this application can be selected from the tailorable thin layer chromatography plates of Tianjin Silida Technology Co., Ltd., or prefabricated silica gel G plates manufactured by Qingdao Yuminyuan or Qingdao Ocean Chemical Plant Branch. Results demonstrate that this method is robust and meets identification requirements. The cigarette volume used for thin layer chromatography testing in this application is 5-15 μL, specifically 8-14 μL, and more specifically 8 μL, 10 μL, 12 μL, or 14 μL.

[0055] In order to make the fluorescent spots appear clearly, the dried thin layer plate was sprayed with 10% sulfuric acid ethanol solution before inspection under ultraviolet light, and then heated at 105°C.

[0056] The inspection temperature described in the present application is 4°C to 35°C, and the thin layer identification method provided by the present invention has good adaptability to different temperatures.

[0057] The humidity of the inspection of the present invention is 32% rh to 75% rh. The thin layer identification method provided by the present invention has good adaptability to different humidity.

[0058] In this application, when identifying flaxseed medicinal materials, a test solution and a flaxseed control medicinal material solution are prepared separately. These two solutions are then spotted on the same silica gel G thin layer for thin-layer chromatography and inspection to determine whether they are flaxseed medicinal materials. When identifying flaxseed formula granules, a test solution and a flaxseed control medicinal material solution are prepared separately. These two solutions are then spotted on the same silica gel G thin layer for thin-layer chromatography and inspection to determine whether they are flaxseed formula granules.

[0059] The present application provides a thin-layer chromatography (TLC) identification method for flaxseed, comprising the following steps: S1) extracting and dissolving the flaxseed to be tested to obtain a test solution, wherein the flaxseed to be tested is a flaxseed medicinal material or a flaxseed granule; S2) extracting and dissolving a flaxseed control medicinal material to obtain a control medicinal material solution; S3) subjecting the test solution and the control medicinal material solution to thin-layer chromatography, using a silica gel G TLC plate and a developing solvent of petroleum ether-ethyl acetate-glacial acetic acid; S4) inspecting under ultraviolet light. If a fluorescent spot of the same color appears in the chromatogram of the test sample at a position corresponding to that of the control medicinal material, the flaxseed medicinal material or the flaxseed granule is identified. The present invention aims to establish a TLC identification method for flaxseed, which can be used to quickly and effectively identify flaxseed medicinal materials or their preparations by observing whether the fluorescent spots of the same color appear in the TLC of the test sample and the control medicinal material at the same position, thereby filling the gap in the qualitative identification of flaxseed, improving the quality standards of flaxseed medicinal materials and their preparations, and ensuring drug safety.

[0060] In order to further understand the present invention, the thin layer identification method of flax seeds provided by the present invention is described in detail below with reference to the examples. The protection scope of the present invention is not limited by the following examples.

[0061] The instruments and reagents used in the following examples are specifically as follows:

[0062] 1.1 Instrument

[0063] Heating plate, mortar, and thin layer imaging system: CAMAG TLC Visualizer, silica gel GF254 thin layer plate (Qingdao Ocean Chemical Plant, batch number: 20180527, Tianjin Silida Technology Co., Ltd., batch number: 191016, Merck, batch number: HX87183353);

[0064] 1.2 Reagents

[0065] Chloroform, anhydrous ethanol, n-butanol, toluene, ethyl acetate, methanol, glacial acetic acid, formic acid, all of the above reagents were of analytical grade, and water was ultrapure water;

[0066] 1.3 Drug testing

[0067] Flaxseed control medicinal material (Chengdu Pusi Biotechnology Co., Ltd., batch number: PS030308); flaxseed medicinal materials (XXLS202309291, XXLS2023092921, XXLS202309292); flaxseed formula granules (batch numbers: B2310028, B2310029, B23109030);

[0068] Example 1 Thin layer method establishment

[0069] 1.1 Preparation of test solution

[0070] Take 2g of linseed powder, add 100ml of water, boil for 30 minutes, filter, concentrate the filtrate to 20ml, add 2ml of hydrochloric acid, heat and reflux for 2 hours, evaporate to dryness, dissolve the residue in 10ml of water, shake and extract with ethyl acetate 3 times, each time 10ml, combine the ethyl acetate solutions, evaporate to dryness, and dissolve the residue in 1ml of methanol to prepare the test solution;

[0071] 1.2 Preparation of control medicinal material solution

[0072] Take 3g of linseed control medicinal material and prepare control medicinal material solution according to the method of test solution;

[0073] 1.3 Assay

[0074] According to the thin layer chromatography method (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia), 10 μl of each of the above two solutions were taken and spotted on the same silica gel G thin layer plate. The plate was developed with petroleum ether (60-90°C)-ethyl acetate-glacial acetic acid (10:1:0.1) as the developing solvent. The plate was taken out and dried in the air. It was sprayed with 10% ethanolic sulfuric acid solution and heated at 105°C until the spots were clearly colored. The plate was then inspected under ultraviolet light (365 nm). If a fluorescent spot of the same color appeared in the chromatogram of the test sample at the corresponding position in the chromatogram of the control medicinal material, it was the flaxseed medicinal material.

[0075] Example 2 Methodological Investigation

[0076] 2.1 Sampling quantity inspection

[0077] Under the experimental conditions proposed above, 8μl, 10μl, 12μl, and 14μl of flaxseed control solution and 8μl, 10μl, 12μl, and 14μl of flaxseed solution were spotted on the same silica gel G thin layer plate, and the plate was developed according to the proposed thin layer chromatography conditions and examined. The results are as follows Figure 1 As stated, Figure 1 The figures are thin layer chromatograms of flaxseed control medicinal material and flaxseed medicinal material with different sample amounts. Among them, 1 to 4 are thin layer chromatograms of flaxseed control medicinal material, and 5 to 8 are thin layer chromatograms of flaxseed medicinal material. It can be seen from the figure that when 8 to 14 μl of the control medicinal material solution and the test solution are sampled, the thin layer chromatogram fluorescent spots are clearly colored, and it is determined that the sample amount of the flaxseed control medicinal material solution and the flaxseed medicinal material is 8 to 14 μl; in the test sample chromatogram, fluorescent spots of the same color are displayed at the corresponding positions in the control medicinal material chromatogram.

[0078] 2.2 Specificity Investigation

[0079] According to the above test sample preparation method, negative solution, flaxseed control medicinal material solution and flaxseed medicinal material solution were prepared respectively, and spotted on the same thin layer plate respectively. The plate was developed according to the proposed thin layer chromatography conditions and examined. The results were as follows: Figure 2 As shown, Figure 2 The thin layer chromatograms of negative solution, flax seed control medicinal material and flax seed medicinal material are shown in Figure 1. The thin layer chromatogram of negative solution, 2. The thin layer chromatogram of flax seed control medicinal material, and 3. The thin layer chromatogram of flax seed medicinal material. Figure 2 It can be used as a TLC specificity test chart for flaxseed medicinal materials. As can be seen from the figure, the negative sample has no interference with the flaxseed medicinal material test sample. The method has good specificity and can be used to identify flaxseed medicinal materials.

[0080] 2.3 Durability Investigation

[0081] 2.3.1 Comparison of different thin layer plates

[0082] The tailorable thin layer chromatography plates of Tianjin Silida Technology Co., Ltd., prefabricated silica gel G plates of Qingdao Yuminyuan and Qingdao Ocean Chemical Plant Branch were selected and tested according to the proposed test methods, such as Figures 3-5 As shown, Figure 3 This is a thin layer chromatogram under the Tianjin Silida thin layer chromatography plate, where 1 is the thin layer chromatogram of the flaxseed control medicinal material, and 2 to 4 are thin layer chromatograms of the flaxseed medicinal material. Figure 4 This is a thin layer chromatogram under the Qingdao Yuminyuan thin layer chromatography plate, where 1 is the thin layer chromatogram of the flaxseed control medicinal material, and 2 to 4 are thin layer chromatograms of the flaxseed medicinal material. Figure 5The thin layer chromatograms under the Qingdao Ocean Thin Layer Chromatography Plate, among which 1 is the thin layer chromatogram of flaxseed control medicinal material, 2 to 4 are the thin layer chromatograms of flaxseed medicinal material; Figures 3-5 It can be seen that the three brands of thin-layer plates can all meet the identification requirements, indicating that the identification method has good durability; 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.

[0083] 2.3.2 Comparison of different temperatures

[0084] Take the spotted thin layer plate and develop it at low temperature of 4℃ and high temperature of 35℃ respectively. Figure 6 and Figure 7 As shown, Figure 6 The figures are the thin layer chromatograms of flaxseed control medicinal material and flaxseed medicinal material at 4℃, among which 1 is the thin layer chromatogram of flaxseed control medicinal material, and 2 to 4 are the thin layer chromatograms of flaxseed medicinal material. Figure 7 The thin layer chromatograms of flaxseed reference medicinal material and flaxseed medicinal material at 35℃ are shown in Figure 1, where 1 is the thin layer chromatogram of flaxseed reference medicinal material, and 2 to 4 are the thin layer chromatograms of flaxseed medicinal material. Figures 6-7 It can be seen that this method has good durability at 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.

[0085] 2.3.3 Comparison of different humidity levels

[0086] Take the spotted thin layer plate and develop it in a humidity environment of 32% rh and 75% rh, respectively. Figures 8-9 As shown, Figure 8 The figures are the thin layer chromatograms of 32% rh flaxseed control medicinal material and flaxseed medicinal material, wherein 1 is the thin layer chromatogram of flaxseed control medicinal material, and 2 to 4 are the thin layer chromatograms of flaxseed medicinal material. Figure 9 The figures are thin layer chromatograms of 75% rh flaxseed control medicinal material and flaxseed medicinal material. Figure 1 is the thin layer chromatogram of the flaxseed control medicinal material, and figures 2 to 4 are the thin layer chromatograms of the flaxseed medicinal material. It can be seen from the figures that the 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.

[0087] 2.3.4 Medicinal Material Verification

[0088] Three batches of flaxseed medicinal materials were subjected to thin layer chromatography identification and verification. The experimental results are as follows Figure 10 As shown, Figure 10 The thin layer chromatograms of different batches of flaxseed medicinal materials and flaxseed control medicinal materials are shown in Figure 1. 1 is the thin layer chromatogram of flaxseed control medicinal materials, and 2 to 4 are the thin layer chromatograms of flaxseed medicinal materials of batches XXLS202309291, XXLS2023092921, and XXLS202309292, respectively. Figure 10 It can be seen that 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.

[0089] Example 3

[0090] Formula particle verification

[0091] 3.1 Preparation of test solution

[0092] Take 2g of flaxseed granules, grind them into powder, add 20ml of water and 2ml of hydrochloric acid, heat and reflux to extract for 2 hours, evaporate to dryness, dissolve the residue in 10ml of water, shake and extract with ethyl acetate 3 times, each time 10ml, combine the ethyl acetate solutions, evaporate to dryness, and dissolve the residue in 1ml of methanol to prepare the test solution;

[0093] 3.2 Preparation of control medicinal material solution

[0094] Take 3g of linseed control medicinal material and prepare control medicinal material solution according to the method of test solution;

[0095] 3.3 Assay

[0096] According to the thin layer chromatography method (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia), 10 μl of each of the above two solutions were taken and spotted on the same silica gel G thin layer plate. The plate was developed with petroleum ether (60-90°C)-ethyl acetate-glacial acetic acid (10:1:0.1) as the developing solvent. The plate was taken out and dried in the air. It was sprayed with 10% ethanolic sulfuric acid solution and heated at 105°C until the spots were clearly colored. The plate was then inspected under ultraviolet light (365 nm). If a fluorescent spot of the same color appeared in the chromatogram of the test sample at the corresponding position in the chromatogram of the control medicinal material, it was the flaxseed medicinal material.

[0097] The results of the thin layer chromatography identification and verification experiments on three batches of flaxseed formula particles are as follows: Figure 11 As shown, Figure 11 The figures are the thin layer chromatograms of different batches of flaxseed formula granules and flaxseed control medicinal materials. Among them, 1 is the thin layer chromatogram of the flaxseed control medicinal material, and 2 to 4 are the thin layer chromatograms of flaxseed formula granules from batches B2310028, B2310029, and B23109030, respectively. The results show that 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.

[0098] Comparative Example 1

[0099] Take 2g of linseed powder, add 25ml of ethyl acetate, ultrasonicate for 30min, filter, evaporate the filtrate to dryness, and dissolve the residue in 1ml of ethyl acetate to prepare the test solution; take 3g of linseed control medicinal material, add 100ml of water, decoct for 30min, filter, evaporate the filtrate to dryness, and prepare the control medicinal material solution by the same method starting from "add 25ml of ethyl acetate"; according to the thin layer chromatography method (General Rules 0502 of the 2020 edition of the Chinese Pharmacopoeia), take 15μl of each of the above two solutions and spot them on the same silica gel G thin layer plate, use toluene-ethyl acetate-formic acid (10:1:0.2) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and observe under ultraviolet light (365nm); the experimental results are as follows Figure 12 As shown, Figure 12 The thin layer chromatograms of linseed control medicinal materials and linseed medicinal materials are shown in Figures 1 to 3. Figures 4-6 The results of thin layer chromatography of flaxseed medicinal materials showed that using the above thin layer identification method, there were no obvious spots between the flaxseed control medicinal materials and the medicinal materials, and the two could not be identified.

[0100] Comparative Example 2

[0101] Take 2g of linseed medicinal material powder, add 20ml of water to dissolve, shake and extract with ethyl acetate twice, 20ml each time, combine the ethyl acetate solution, evaporate to dryness, add 1ml of methanol to dissolve the residue, and use it as the test solution; take 3g of linseed control medicinal material, add 100ml of water, decoct for 30min, filter, and concentrate the filtrate to about 20ml. 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), take 20μl of each of the above two solutions and spot them on the same silica gel G thin layer plate, use toluene-acetone-glacial acetic acid (8:3:0.3) as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and observe under ultraviolet light (365nm); the experimental results are as follows Figure 13 As shown, Figure 13 The thin layer chromatograms of linseed reference medicinal material and linseed medicinal material are shown in Figures 1 to 2. Figures 3-4 The results of thin layer chromatography of flaxseed medicinal materials showed that using the above thin layer identification method, there were no obvious spots between the flaxseed control medicinal materials and the medicinal materials, and the two could not be identified.

[0102] The above embodiments are only intended to help understand the method and core concept of the present invention. It should be noted that, without departing from the principles of the present invention, a number of improvements and modifications may be made to the present invention by those skilled in the art, and such improvements and modifications also fall within the scope of protection of the claims of the present invention.

[0103] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A thin layer identification method for flax seeds, comprising the following steps: S1) extracting and dissolving flaxseed to be tested to obtain a test solution, wherein the flaxseed to be tested is flaxseed medicinal material or flaxseed formula granules; S2) extracting and dissolving the flaxseed control medicinal material to obtain a control medicinal material solution; S3) subjecting the test solution and the control medicinal material solution to thin layer chromatography; the thin layer plate is a silica gel G thin layer plate, and the developing solvent is petroleum ether-ethyl acetate-glacial acetic acid; the volume ratio of the petroleum ether, the ethyl acetate, and the glacial acetic acid is (9-11):1:(0.08-0.12); S4) Place the sample under ultraviolet light for inspection. If a fluorescent spot of the same color appears in the chromatogram of the test sample at the corresponding position of the control herbal medicine, it is the flaxseed herbal medicine or flaxseed formula granules. Step S1) is specifically as follows: When the flaxseed to be tested is linseed medicinal material, the linseed is decocted with water and then filtered, the filtrate is concentrated and then hydrochloric acid is added, and the mixture is heated under reflux for extraction, and the residue is evaporated to dryness and dissolved in water, and then extracted with ethyl acetate, and the residue is evaporated to dryness and dissolved in methanol to obtain a test solution; When the flaxseed to be tested is a flaxseed formula granule, the flaxseed formula granule is ground, and then water and hydrochloric acid are added, and reflux extraction is performed. After evaporation, water is added to the residue, and then extraction is performed with ethyl acetate. After evaporation again, methanol is added to dissolve the residue to obtain a test solution.

2. The thin layer identification method according to claim 1, characterized in that: The ratio of the linseed, the hydrochloric acid and the ethyl acetate is 1 g: (1-3) ml: (2-8) ml.

3. The thin layer identification method according to claim 1, characterized in that: Step S2) is specifically as follows: The linseed control medicinal material was decocted with water and filtered. The filtrate was concentrated and then added with hydrochloric acid. The mixture was heated under reflux for extraction. After evaporation, the residue was dissolved in water and extracted with ethyl acetate. The residue was evaporated and dissolved in methanol to obtain the test solution. The ratio of the linseed reference medicinal material, the hydrochloric acid, and the ethyl acetate is 1 g: (1-3) ml: (2-8) ml.

4. The thin layer identification method according to claim 1, characterized in that: In step S3), the sample volume for thin layer chromatography detection is 5-15 μL.

5. The thin layer identification method according to claim 1, characterized in that: The volume ratio of the petroleum ether, the ethyl acetate and the glacial acetic acid is 10:1:0.

1.

6. The thin layer identification method according to claim 1, characterized in that: The wavelength of the ultraviolet lamp is 365nm.

7. The thin layer identification method according to claim 1, characterized in that: The review also includes: The developed thin layer plate was sprayed with 10% sulfuric acid ethanol solution and then heated at 100-110°C.

8. The thin layer identification method according to claim 1, characterized in that: The inspection temperature is 4°C to 35°C.

9. The thin layer identification method according to claim 1, characterized in that: The humidity under inspection is 32% rh ~ 75% rh.

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