A thin layer identification method of plum blossom formula granules

Abstract

The present application provides a kind of plum blossom formula granule thin layer identification method, comprising the following steps: taking plum blossom formula granule, ultrasonic treatment with methanol, filtering, filter liquor is evaporated, residue is dissolved with methanol, to obtain plum blossom formula granule solution;Plum blossom formula granule solution is drawn and is spotted on silica gel G thin layer plate prepared with sodium acetate solution, using developing agent development, take out, air dry, spray color reagent heating, under UV light inspection.The method of the present application can quickly and effectively identify plum blossom, the method has high precision and sensitivity, good stability, can accurately identify plum blossom formula granule, and has good application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine identification, in particular to a thin-layer identification method for plum blossom formula granules. Background Art

[0002] Plum Blossom Formula Granules are processed from the dried flower buds of Prunus mume (Sieb) Sieb. et Zucc., a plant in the Rosaceae family. Medicinal plum blossoms are harvested in early spring before they bloom and promptly dried at low temperatures. They soothe the liver and stomach, resolve phlegm, detoxify, and disperse stagnation. They are primarily used to treat liver and stomach pain, restlessness, depression, and scrofula. Plum blossoms are primarily produced in Jiangsu, Zhejiang, and Anhui provinces. The main chemical components of plum blossoms include flavonoids, phenylpropanoids, and organic acids.

[0003] The principle of thin-layer chromatography is to utilize the differences in the affinity of different substances for adsorbents and developing agents, resulting in the separation of substances and achieving separation. Currently, Meihua uses polyamide film for thin-layer chromatography development in the thin-layer identification items in the pharmacopoeia and the formula granule standards of multiple provinces. The principle of polyamide film is the formation of hydrogen bonds between polyamide and the separated substances. The strength of the hydrogen bonds formed by different substances leads to the separation of substances and achieves separation. The principle of silica gel G thin-layer plate is that the separated substances have different adsorption effects between silica gel and developing agents, resulting in the separation of substances and achieving separation. Polyamide film is relatively expensive, soft and easy to bend, which makes it slightly inconvenient to develop, inspect and take pictures.

[0004] Currently, there is no published method for using silica gel G thin layer plates to perform thin layer identification testing of plum blossoms or plum blossom formula particles. Summary of the Invention

[0005] The present invention provides a thin-layer identification method for plum blossom formula granules. The method can quickly and effectively identify plum blossoms, save costs, and is more convenient for unfolding, inspecting and photographing. The method has high precision and sensitivity, good stability, and can accurately identify plum blossom formula granules.

[0006] The present invention provides a thin layer identification method for plum blossom formula granules, which is characterized by comprising the following steps:

[0007] Take the plum blossom formula granules, add methanol for ultrasonic treatment, filter, evaporate the filtrate to dryness, and dissolve the residue in methanol to obtain a plum blossom formula granule solution;

[0008] Pipette the plum blossom formula granule solution onto a silica gel G thin layer plate prepared with sodium acetate solution, develop it with a developing agent, take it out, dry it, spray it with a color developer, heat it, and inspect it under ultraviolet light.

[0009] Preferably, the weight-to-volume ratio g / ml of the plum blossom formula particles and methanol is 1:250.

[0010] Preferably, the ultrasonic treatment power is 250W, the frequency is 53kHZ, and the time is 30 minutes.

[0011] Preferably, the plum blossom formula granule solution is a solution containing 0.05g / ml of granules.

[0012] Preferably, the weight / volume percentage of the sodium acetate solution is 4%.

[0013] Preferably, the developing agent is composed of ethyl acetate, butanone, formic acid and water.

[0014] Preferably, the volume ratio of the ethyl acetate, butanone, formic acid and water is 5:3:1:1.

[0015] Preferably, the color developing agent is a 3% weight / volume percentage aluminum chloride ethanol solution, the heating is heating at 40℃ for 3 minutes, and the wavelength of the ultraviolet light lamp is 365nm.

[0016] The present application has the following beneficial effects:

[0017] The thin layer identification method of the plum blossom formula granule adopted by the present application can save costs compared to the traditional polyamide film, and is more convenient for development and viewing and photographing by using a 4% sodium acetate silica gel G thin layer plate; in addition, the thin layer chromatography identification method can better separate the effective components by using the unique developing agent components and component formula.

[0018] In summary, the method of the present application can quickly and effectively identify plum blossom by using a 4% sodium acetate silica gel G thin layer plate + developing agent combination, and has high precision and sensitivity, good stability, and can accurately identify plum blossom formula granules, and has good application prospects. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 is a thin layer plate after taking a sample, which is developed in a temperature environment of low temperature 8.5℃. 1-4 are formula granules, 5 is a control medicinal material, and 6 is a control product.

[0020] Figure 2 is a thin layer plate after taking a sample, which is developed in a temperature environment of normal temperature 26℃. 1-4 are formula granules, 5 is a control medicinal material, and 6 is a control product.

[0021] Figure 3 is a thin layer plate after taking a sample, which is developed in a humidity environment of 40%. 1-4 are formula granules, 5 is a control medicinal material, and 6 is a control product.

[0022] Figure 4The thin layer identification method of the plum blossom formula granules of the present invention is adopted, and the thin layer plate after the sample is taken is developed in an environment of 85% humidity. Among them, 1 to 4 are formula granules, 5 is a control medicinal material, and 6 is a reference substance.

[0023] Figure 5 The thin layer identification method of the plum blossom formula granules of the present invention is adopted, which uses the German MercK ordinary silica gel G plate. Among them, 1 to 4 are formula granules, 5 is a control medicinal material, and 6 is a reference substance.

[0024] Figure 6 The thin layer identification method of the plum blossom formula granules of the present invention is adopted, which uses the German MercK silica gel high-efficiency G plate. Among them, 1 to 4 are formula granules, 5 is a control medicinal material, and 6 is a reference substance.

[0025] Figure 7 The thin layer identification method of the plum blossom formula granules of the present invention is adopted, and the silica gel G plate is used. 1 to 4 are formula granules, 5 is a control medicinal material, and 6 is a reference substance. DETAILED DESCRIPTION

[0026] In order to better illustrate the problem to be solved, the technical solution adopted and the beneficial effects achieved by the technical solution of the present invention, it is now further described in conjunction with specific implementation methods. It is worth noting that the technical solution of the present invention includes but is not limited to the following implementation methods.

[0027] If no specific techniques or conditions are specified in the examples of the present invention, the techniques or conditions described in the literature in the field or the product instructions shall be followed. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0028] Apparatus and Materials:

[0029] ATS 4 TLC automatic sample analyzer (CAMAG, Switzerland), TLC double-slot developing cylinder, TLCvisualizer (CAMAG, Switzerland), balance (Sartorius, Germany), and thin layer silica gel precast plate (Merck).

[0030] Methanol, ethyl acetate, butanone, formic acid, ethanol, and aluminum chloride were all analytical grade (Guangzhou Chemical Reagent Factory).

[0031] The ratio of the developing solvent to the mobile phase in the present invention is a volume ratio.

[0032] The plum blossom reference medicinal material of the present invention: China Food and Drug Inspection Institute, batch number 121656-201802.

[0033] The plum blossom formula granules of the present invention: Guangdong Yifang Pharmaceutical Co., Ltd., batch numbers CG2011006, CG2011007, and CG2011008.

[0034] The rutin reference substance of the present invention is: China Food and Drug Inspection Institute, batch number 100080-202012.

[0035] The silica gel G thin layer plate prepared with 4% sodium acetate solution of the present invention is as follows: 4g sodium acetate is dissolved in 100ml water to obtain 4% sodium acetate solution, a silica gel G thin layer plate is placed in the 4% sodium acetate solution to soak, taken out, and dried to obtain a silica gel G thin layer plate prepared with 4% sodium acetate solution.

[0036] The preparation method of the 3% aluminum chloride ethanol solution of the present invention is as follows: 3g AlCl3 is dissolved in 97g ethanol.

[0037] Example 1

[0038] This embodiment provides a thin layer identification method for plum blossom formula granules, comprising the following steps:

[0039] 1. Sample Preparation

[0040] Preparation of control medicinal material solution: Take 0.25g of plum blossom control medicinal material, add 60ml of water, boil for 30 minutes, filter through medium-speed double-circle qualitative filter paper (pore size 30-50 microns), evaporate the filtrate to dryness, add 25ml of methanol to the residue, and ultrasonically treat for 30 minutes with a power of 250W and a frequency of 53kHz, filter through medium-speed double-circle qualitative filter paper (pore size 30-50 microns), evaporate the filtrate to dryness, and add 2ml of methanol to dissolve the residue.

[0041] Preparation of formula granule solution: Take an appropriate amount of plum blossom formula granules, grind them into powder, take 0.1g, add 25ml of methanol, ultrasonically treat for 30 minutes, with a power of 250W and a frequency of 53kHz, filter through medium-speed double-circle qualitative filter paper (pore size 30-50 microns), evaporate the filtrate to dryness, and add 2ml of methanol to dissolve the residue.

[0042] Preparation of reference solution: Take an appropriate amount of rutin reference substance, add methanol, and dissolve it by ultrasonication for 30 minutes at a power of 250W and a frequency of 53kHz to prepare a solution containing 1mg per ml.

[0043] 2. Chromatographic conditions

[0044] Spotting: 1 μl each of the formula granule solution, control medicinal material solution and reference substance solution were spotted on the same silica gel G thin layer plate prepared with 4% sodium acetate solution.

[0045] Developing solvent: ethyl acetate-butanone-formic acid-water (5:3:1:1)

[0046] Color Development and Inspection: After development, remove the sample, evaporate the solvent, spray with 3% aluminum chloride in ethanol, heat on a 40°C hot plate for 3 minutes, and observe the chromatogram under a UV lamp (365 nm). Photograph and record the TLC image. Fluorescent spots of the same color will appear in the chromatogram of the formula granules at positions corresponding to those in the chromatograms of the control medicinal material and the reference substance.

[0047] Example 2

[0048] Comparison of different temperatures: Take the thin layer plate after spotting in Example 1 and develop it at a low temperature of 8.5℃ and a room temperature of 26℃. This shows that this method has good adaptability to different temperatures. Figure 1-2 .

[0049] Example 3

[0050] Comparison of different humidity: Take the thin layer plate after spotting in Example 1 and develop it in 40% and 85% humidity environment respectively. Figure 3-4 , indicating that this method has good adaptability to different humidity.

[0051] Example 4

[0052] Comparison of silica gel G plates from different manufacturers: German MercK thin layer chromatography plates (silica gel G plates and silica gel high-efficiency G plates) and Qingdao Ocean Chemical Co., Ltd. (silica gel G plates) were selected and tested according to the test method in Example 1. The results are shown in Figures 5 to 7 The results show that different plates can achieve good separation effects.

Claims

1. A thin layer identification method for plum blossom formula granules, characterized in that: The steps include: Take the plum blossom formula granules, add methanol for ultrasonic treatment, filter, evaporate the filtrate to dryness, and dissolve the residue in methanol to obtain a plum blossom formula granule solution; Preparation of reference solution: Take an appropriate amount of rutin reference substance and add methanol to obtain a reference solution; Pipette the Meihua formula granule solution and the reference substance solution onto a silica gel G thin layer plate prepared with sodium acetate solution, develop it with a developing agent, take it out, dry it, spray it with a color developer, heat it, and inspect it under ultraviolet light; The developing agent consists of ethyl acetate, butanone, formic acid and water; The volume ratio of ethyl acetate, butanone, formic acid and water is 5:3:1:1; The color developer is a 3% by weight volume aluminum chloride ethanol solution.

2. The thin layer identification method of plum blossom formula granules according to claim 1, characterized in that: The weight-to-volume ratio of the plum blossom formula particles to methanol is 1:250 g / ml.

3. The thin layer identification method of plum blossom formula granules according to claim 1, characterized in that: The ultrasonic treatment was performed with a power of 250 W, a frequency of 53 kHz, and a duration of 30 minutes.

4. The thin layer identification method of plum blossom formula granules according to claim 1, characterized in that: The plum blossom formula granule solution is a solution containing 0.05 g / ml of granules.

5. The thin layer identification method of plum blossom formula granules according to claim 1, characterized in that: The weight volume percentage of the sodium acetate solution is 4%.

6. The thin layer identification method of plum blossom formula granules according to claim 1, characterized in that: The heating is performed at 40° C. for 3 minutes, and the wavelength of the ultraviolet lamp is 365 nm.

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