阅读说明：本技术 一种紫地榆颗粒及其标煎液干粉的快速多信息薄层鉴别方法 (Rapid multi-information thin-layer identification method for Geranium strictipes granules and target decoction dry powder thereof ) 是由 牛丽颖 田宇柔 吴玲芳 麻景梅 王鑫国 于 2021-09-23 设计创作，主要内容包括：本发明涉及一种紫地榆颗粒及标煎液干粉的快速多信息薄层鉴别方法。其特征在于：用简便、快捷的前处理方法得到供试品与紫地榆药材溶液,分别点于同一块薄层板上,展开后,在3种不同的检视条件下,检视到紫地榆颗粒及标煎液干粉的8种信息斑点,归属于约7种不同的化学成分。荧光斑点、无荧光斑点、喷雾硫酸乙醇溶液后,呈色的颜色斑点,不同的斑点代表着不同的化学成分,斑点清晰,易于质量评价,并做到了不同检视条件下的信息斑点互补,提升了质量检测指标与水准。目前还未查阅到有关紫地榆颗粒及标煎液干粉快速多信息薄层鉴别方法的报道。具新颖性、创新性和实用性。且方法简便、快捷,效率高、成本低、绿色环保。(The invention relates to a rapid multi-information thin-layer identification method for Geranium strictipes granules and standard decoction dry powder. The method is characterized in that: a test sample and a Geranium strictipes medicinal material solution are obtained by a simple and rapid pretreatment method, are respectively spotted on the same thin-layer plate, and 8 information spots of Geranium strictipes particles and standard decoction dry powder are inspected under 3 different inspection conditions after the materials are unfolded, wherein the information spots belong to about 7 different chemical components. Fluorescent spots, no fluorescent spots, colored spots after spraying sulfuric acid ethanol solution, different spots representing different chemical components, clear spots, easy quality evaluation, information spot complementation under different inspection conditions, and improved quality detection index and standard. At present, reports about a rapid multi-information thin-layer identification method for Geranium strictipes granules and standard decoction dry powder are not found. Has novelty, innovation and practicability. And the method is simple, convenient and quick, has high efficiency and low cost, and is green and environment-friendly.)

1. A rapid multi-information thin-layer identification method for Geranium strictipes granules and target decoction dry powder thereof is characterized in that:

weighing Geranium strictipes granules and standard decoction dry powder thereof 0.2g respectively, grinding, adding 70% methanol 2mL respectively, performing ultrasonic treatment for 10 min, centrifuging, and collecting supernatant as respective test solution; taking 0.5g of Geranium strictipes reference medicinal material, adding 3mL of 70% methanol, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a reference medicinal material solution; respectively sucking the three solutions by 6-7 μ l, and respectively dropping on the same silica gel GF₂₅₄Developing with chloroform-ethyl acetate-methanol-concentrated ammonia solution as developer in the volume ratio of 3 to 2 to 3 to 0.5, taking out, air drying,inspecting under 365nm ultraviolet lamp to show main fluorescent spots of the same color in the chromatogram of the test solution at the positions corresponding to those in the chromatogram of the reference solution; inspecting under 254nm ultraviolet lamp to show main spots of the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the reference medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, and inspecting in dark room through lamplight to show main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the control medicinal material.

2. The method for rapidly identifying the geranium strictipes granules and the standard decoction dry powder by the multi-information thin layer as claimed in claim 1, wherein each 1g of granules is equal to 6.1g of crude drugs; each 1g of standard decoction is equivalent to 6.1g of the original medicinal material.

3. The method of claim 1, wherein 8 kinds of information spots belonging to 7 kinds of chemical components are detected on the Geranium strictipes granules and the standard decoction dry powder under 3 different detection conditions using the same developing agent on the same thin layer plate.

Technical Field

The invention relates to a rapid multi-information thin-layer identification method for Geranium strictipes granules and target decoction dry powder thereof. Belongs to the field of thin-layer identification of traditional Chinese medicines. Namely, thin-layer chromatography is adopted for identification, and 8 information spots of the Geranium strictipes granules and the standard decoction dry powder thereof are inspected on the same thin-layer plate under 3 different inspection conditions and belong to about 7 different chemical components.

Background

The so-called fast thin layer identification method is simple, fast and efficient as the name implies, the sample solution and the reference medicinal material solution do not need repeated purification treatment, and no environmental pollution operation procedures such as evaporation, concentration and the like of toxic solvents exist, and after ultrasonic extraction by using a proper solvent, centrifugation, sample application of supernatant, expansion and inspection are mostly carried out, so that various chemical component information spots under various inspection conditions can be obtained.

Geranium strictipes R.Knuth (G.striatum aureus. non Burm. f.) belonging to Geranium of Geraniaceae is used as root for administration, and is named as Dingshan Xiao. Has the effects of clearing heat, promoting diuresis, promoting blood circulation and stopping bleeding. Can be used for treating enteritis, dysentery, dyspepsia, chronic gastritis, menoxenia, and epistaxis; it is used externally to treat traumatic injury. The Ulmus davidiana contains flavonoid glycoside, quercetin and kaempferol, and also contains betulinic acid and gallic acid. The substances rich in the ulmus pumila have the effects of cooling blood, stopping bleeding, detoxifying and healing sores, have good treatment effects on treating hematochezia, scalds and sore toxins, and are good medicines for people with scalds and sore toxins. The gallic acid has antibacterial, antiviral, and antitumor effects. Therefore, the Chinese medicinal composition is widely used in the white nationality, the Yi nationality and other minority regions in Yunnan, and is a commonly used national Chinese medicinal material. At present, the variety of Geranium strictipes is not collected in Chinese pharmacopoeia, and the quality standard collected in local standards is not consulted. The related research on the Geranium strictipes is less, and only the research on the extraction of gallic acid is consulted; gallic acid is taken as an index by some people, and HPLC quantitative determination is carried out; the thin-layer identification is carried out on flavonoid components by people, and the method comprises the following steps: taking 1g of dry Geranium strictipes medicinal material, adding 10ml of methanol, cold soaking overnight, filtering, evaporating filtrate to dryness, and adding 1ml of methanol for dissolving to obtain a sample solution. Preparing another 1g of Geranium strictipes reference medicinal material, and preparing a reference medicinal material solution by the same method. Sucking 1 μ l of each of the above two solutions, respectively dropping on polyamide film, developing with ethanol-water-formic acid (3: 2: 0.1) as developing agent, taking out, air drying, spraying 1% aluminum trichloride ethanol solution for developing color, observing under 365nm ultraviolet lamp, and displaying fluorescent spot with same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the control solution. The identification is carried out by taking a polyamide film as a carrier, and due to the fact that the polyamide film is soft in texture, small in sample loading amount, small in detection information amount, poor in spot definition and inconvenient to operate, the quality evaluation of the medicinal materials and the preparations of the medicinal materials by multiple indexes is difficult to achieve.

Therefore, on the basis of the background technology, aiming at the current situations that the research on the quality detection method of the national traditional Chinese medicine formula particles is relatively lagged, the detection index is low, and the quality supervision is difficult to effectively carry out, in order to improve the convenience, safety and effectiveness of clinical application of patients, the preparation process of the Geranium strictipes particles and the standard decoction dry powder is firstly researched; then, thin-layer identification research is carried out on the particles and the standard decoction dry powder thereof. The method for quickly identifying the multiple information thin layers is simple, convenient and quick, and has no environmental pollution. The preparation process of the garden burnet medicinal material, the granule and the standard decoction dry powder is described as follows:

geranium strictipes medicinal material is prepared by collecting Geranium strictipes according to the processing specification of traditional Chinese medicines, removing impurities, cleaning, slicing, and drying in the sun or at a proper temperature to obtain Geranium strictipes decoction pieces (also called medicinal materials).

Geranium strictipes granules are prepared by decocting pieces of Geranium strictipes in water for 2 times, each time for 1-2 hours, filtering, combining filtrates, concentrating under reduced pressure to an appropriate relative density, spray drying, adding an appropriate amount of dextrin into dried powder, mixing uniformly, and granulating.

The Geranium strictipes standard decoction dry powder is prepared by decocting pieces of Geranium strictipes in water for 2 times, each time for 1-2 hours, filtering, combining filtrates, concentrating at normal temperature to appropriate relative density, adding appropriate amount of dextrin, stirring well, placing in a 60 ℃ oven, oven drying, and pulverizing.

Disclosure of Invention

Firstly creates a rapid multi-information thin-layer identification method for Geranium strictipes granules and standard decoction dry powder. The method is characterized in that: a test sample and a Geranium strictipes medicinal material solution are obtained by a simple and rapid pretreatment method, are respectively spotted on the same thin-layer plate, and 8 information spots of Geranium strictipes particles and standard decoction dry powder are inspected under 3 different inspection conditions after the materials are unfolded, wherein the information spots belong to about 7 different chemical components. Fluorescent spots, no fluorescent spots, colored spots after spraying sulfuric acid ethanol solution, different spots representing different chemical components, clear spots, easy quality evaluation, information spot complementation under different inspection conditions, and improved quality detection index and standard. At present, a rapid multi-information thin-layer identification report of Geranium strictipes granules and standard decoction dry powder is not found. Has novelty, innovation and practicability. And the method is simple, convenient and quick, has high efficiency and low cost, and is green, environment-friendly and pollution-free.

The technical scheme adopted by the invention for solving the technical problems is as follows:

weighing Geranium strictipes granules and dry powder 0.2g respectively, grinding, respectively adding 70% methanol 2mL, performing ultrasonic treatment for 10 min, centrifuging, and collecting supernatant as test solution; taking 0.5g of Geranium strictipes reference medicinal material, adding 3mL of 70% methanol, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a reference medicinal material solution; respectively sucking the three solutions by 6-7 μ l, and respectively dropping on the same silica gel GF₂₅₄Developing on a thin layer plate with chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent at volume ratio of 3: 2: 3: 0.5, taking out, air drying, inspecting under ultraviolet lamp 365nm to obtain main fluorescent spots with the same color in the chromatogram of the sample and the corresponding positions of the chromatogram of the reference medicinal material; inspecting under 254nm ultraviolet lamp to show main spots of the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the reference medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, and inspecting in dark room through lamplight to show main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the control medicinal material.

The principle of the invention is as follows:

according to the chemical structure and properties of each effective component of the traditional Chinese medicine, a test sample and a reference medicinal solution are simply, conveniently and quickly prepared by adopting a proper extraction solvent according to a similar compatible extraction principle. And then proper developing agent is adopted for developing, and various chemical components can be well separated on the thin-layer plate along with the selected developing agent according to different adsorption, desorption, re-adsorption and re-desorption capabilities. By means of effective components with similar polarities, different color spots are displayed on the same thin-layer plate under different inspection conditions, and a thin-layer chromatogram with multiple information is obtained.

The invention has the following innovation points and beneficial effects:

1. firstly creates a rapid multi-information thin-layer identification method for Geranium strictipes granules and standard decoction dry powder. Namely, a test sample and a Geranium strictipes medicinal material solution are obtained by a simple and rapid pretreatment method, and are respectively spotted on the same thin-layer plate, and after the thin-layer plate is unfolded, 8 information spots of Geranium strictipes particles and standard decoction dry powder are inspected under 3 different inspection conditions, wherein the information spots belong to about 7 different chemical components. Fluorescent spots, no fluorescent spots, colored spots after spraying sulfuric acid ethanol solution, different spots representing different chemical components, clear spots, easy quality evaluation, information spot complementation under different inspection conditions, and improved quality detection index and standard. At present, a rapid multi-information thin-layer identification report of Geranium strictipes granules and standard decoction dry powder is not found. Has novelty, innovation and practicability. And the method is simple, convenient and quick, has high efficiency and low cost, is green and environment-friendly, and has no environmental pollution.

2. Developing the Geranium strictipes granules and the standard decoction dry powder by using chloroform-ethyl acetate-methanol-concentrated ammonia test solution with the volume ratio of 3: 2: 3: 0.5 as a developing agent, and inspecting under an ultraviolet lamp of 365nm, wherein 1 bluish violet fluorescent spot is detected in the medicinal materials, the granules and the standard decoction dry powder (figure 1); inspecting under 254nm ultraviolet lamp to obtain medicinal materials and standard decoction powder with 3 dry powders and 4 brown spots, considering that the powder may be caused by instantaneous spray drying and little loss of effective components (FIG. 2); spraying 10% sulphuric acid ethanol solution for color development, inspecting spots under sunlight for poor definition, but inspecting in dark room through lamplight, wherein dry powder of medicinal materials, granules and standard decoction all present 4 brownish red spots, and one Rf value is about 0.2 with high content (FIG. 3); the same thin layer plate can detect 8 information spots under three inspection conditions, and the spot size, shape and Rf value under different inspection conditions are compared to belong to 7 different chemical components.

3. The innovation of the invention is that the selected alkaline developing agent overcomes the defect of tailing of macromolecular tannic acid component under the acidic developing agent. The polarity and the separating capacity reach the expected target, the chemical components of the Geranium strictipes are uniformly distributed in the whole area of the thin layer plate, and the Rf value falls within the range of 0.2-0.7. A total of 8 information spots were observed, representing 7 different chemical components, indicating that the component spots observed were substantially non-overlapping. Such separation is achieved due to the exact pH, composition and ratio of the developing solvent chosen. The method and the device realize the improvement of detection indexes and supervision on the premise of not increasing any workload and detection cost. At present, the same reports are not found, and the method has the practicability of novelty, creativity, resource saving, efficiency improvement and particle quality guarantee.

4. From the analysis of the composition and proportion of the developing agent, the multi-information spots detected by the identification are all water-soluble components.

5. The identification method can obtain the sample and the medicinal material solution only by 0.5g of Geranium strictipes medicinal material powder, 0.2g of particles and standard decoction dry powder respectively and 7ml of methanol in 20 minutes, and can perform multi-index evaluation on the sample quality by adding 12ml of developing agent for 30 minutes and taking 1 hour totally, thereby being simple, convenient and quick and having higher practical value.

Drawings

FIG. 1 is a TLC image of Geranium strictipes granules and their standard decoction powder under 365nm UV light.

FIG. 2 is a TLC image of Geranium strictipes granules and their standard decoction powder under 254nm UV light.

FIG. 3 is a TLC image of the particle of Geranium strictipes and its target decoction powder after being developed with 10% ethanol sulfate solution and inspected by light in a dark room.

FIGS. 1, 2 and 3 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1.2 is Geranium strictipes granules; 3. 4 is the dried powder of the standard decoction of the Geranium strictipes; 5.6 is Geranium strictipes medicinal material;

detailed description of the invention

Weighing Geranium strictipes granules and dry powder 0.2g respectively, grinding, respectively adding 70% methanol 2mL, performing ultrasonic treatment for 10 min, centrifuging, and collecting supernatant as test solution; taking 0.5g of Geranium strictipes reference medicinal material, adding 3mL of 70% methanol, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a reference medicinal material solution; respectively sucking the three solutions by 6-7 μ l, and respectively dropping on the same silica gel GF₂₅₄Developing on a thin layer plate with chloroform-ethyl acetate-methanol-concentrated ammonia solution as developing agent at volume ratio of 3: 2: 3: 0.5, taking out, air drying, inspecting under ultraviolet lamp 365nm to obtain main fluorescent spots with the same color in the chromatogram of the sample and the corresponding positions of the chromatogram of the reference medicinal material; inspecting under 254nm ultraviolet lamp to show main spots of the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the reference medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed, and inspecting in dark room through lamplight to show main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the control medicinal material.