CN113791166A - One-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder thereof - Google Patents
One-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder thereof Download PDFInfo
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- CN113791166A CN113791166A CN202111114093.7A CN202111114093A CN113791166A CN 113791166 A CN113791166 A CN 113791166A CN 202111114093 A CN202111114093 A CN 202111114093A CN 113791166 A CN113791166 A CN 113791166A
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Abstract
The invention relates to a one-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder. The method is characterized in that: a test sample and a five-qi yang-rising herbal medicine solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and after being unfolded, 17 information spots of five-qi yang-rising herbal medicine particles and standard decoction dry powder are detected under 5 detection conditions and belong to about 13 different chemical components. The spots before color development, the fluorescent spots, the non-fluorescent spots, the fluorescent spots after color development of sulfuric acid ethanol solution and the colored spots, different spots represent different chemical components, not only the information spot complementation is exerted, but also the method is simple and convenient, the cost is low, the number of thin layer plates is small, the inspection condition is high, the detection information amount is large, the index can be calculated, and the novelty, the creativity and the practicability are obvious. At present, no report on a one-board multi-information thin-layer identification method of five-qi Korean-yang grass particles and standard decoction dry powder is found.
Description
Technical Field
The invention relates to a one-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder thereof. Belongs to the field of thin-layer identification of traditional Chinese medicines. Namely, by adopting thin-layer chromatography for identification, 17 information spots of the five-qi chaoyang grass granules and the standard decoction dry powder thereof are inspected on the same thin-layer plate under 5 different inspection conditions, and the information spots belong to about 13 different chemical components.
Background
The identification method is characterized in that more than ten kinds of information spots are detected on a thin-layer plate under 5 different detection conditions, the method is simple, convenient, rapid and efficient, the sample solution and the contrast medicinal solution do not need to be purified repeatedly, and environmental pollution operation procedures such as evaporation and concentration of toxic solvents are avoided.
The five-qi chaoyang grass is also called as lubranzheng, collected in the first part of the 2020 th edition of Chinese pharmacopoeia and is dry whole grass of Geum aleppicum Jacq. or Geum japonicum Thunb. var. chinensis Bolle, a Rosaceae plant. Has the functions of replenishing qi to invigorate the spleen, enriching the blood and nourishing yin, moistening the lung and reducing phlegm. Can be used for treating deficiency of both qi and blood, consumptive disease, cough, spleen deficiency, and leukorrhagia. Is a common ethnic Chinese medicinal material. Mainly contains chemical components such as triterpenes, flavonoids, tannins, lignins, volatile oil and the like. In addition, there are some fatty acids, phenols, saccharides, and the like. Although the chemical components contained in the method are various, the thin-layer identification research reports are few, the thin-layer development is carried out by adopting a method of Chinese pharmacopoeia and using gallic acid as a detection index, and the detailed identification method is as follows:
collecting powder of herba Gei Piloselloidis (herba Gei Piloselloidis) 1g, adding diluted ethanol 20ml, ultrasonic extracting for 30 min, filtering, evaporating filtrate to dryness, dissolving residue with water 10ml, washing with chloroform for 2 times (10 ml each time), discarding chloroform solution, adding ethyl acetate into water solution, shaking and extracting for 2 times (10 ml each time), mixingThe ethyl acetate solution was evaporated to dryness, and 2ml of acetone was added to the residue to dissolve it, thereby obtaining a sample solution. Taking 1g of herba Bluellae positive control medicinal material, and making into control medicinal solution by the same method. Adding acetone into gallic acid control to obtain solution containing 1mg of gallic acid per 1ml, and making into control solution. Sucking the three solutions 2-3 mul each, and respectively dropping on the same silica gel GF254Spreading chloroform-ethyl formate-formic acid (5: 1) as developing agent on the thin layer plate, taking out, air drying, and inspecting under ultraviolet lamp (254 nm). Spots of the same color appear on the chromatogram of the test solution at the positions corresponding to those on the chromatograms of the reference medicinal material and the reference solution. The method is complicated and time-consuming, the operation needs 2.5 hours, the extraction and development of the solvent is 130ml, most importantly, only one component of the gallic acid is detected, and the gallic acid is a common component in various plants and has poor specificity.
Aiming at the current situation that the research of the national traditional Chinese medicine formula particle quality detection method is relatively lagged, the detection index is low, and the quality supervision is difficult to effectively carry out, and in order to be suitable for the convenience, safety and effectiveness of clinical application of patients, the preparation process of five-qi Korean monochoria herb particles and standard decoction dry powder is firstly researched; then, the special thin-layer identification research is carried out on the particles and the standard decoction dry powder. The method for identifying the multiple information thin layers on one plate is simple, convenient and quick, and has no environmental pollution. The preparation process of the five-qi Korean-yang herb medicinal material, the granules and the standard decoction dry powder is described as follows:
the five-qi yang-rising herbal medicine material is prepared by collecting five-qi yang-rising grass according to the processing specification of traditional Chinese medicines, removing impurities, cleaning, cutting into sections, and drying in the sun or at a proper temperature to obtain five-qi yang-rising grass decoction pieces (also called medicinal materials).
The five-qi Korean-herb granule is prepared by decocting five-qi Korean-herb decoction pieces with 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 five-qi Korean-herb-labeled decoction dry powder is prepared by decocting five-qi Korean-herb decoction pieces with water for 2 times, 1-2 hours each time, filtering, combining filtrates, concentrating at normal temperature to appropriate relative density, adding an appropriate amount of dextrin, stirring uniformly, placing in a 60 ℃ oven, drying, and pulverizing.
Disclosure of Invention
Firstly creates a one-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder. Namely, a test sample and a five-qi yang-rising herbal medicine solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and after being unfolded, 17 information spots of five-qi yang-rising herbal granules and standard decoction dry powder are detected under 5 detection conditions and belong to about 13 different chemical components. The spots before color development, the fluorescent spots, the non-fluorescent spots, the fluorescent spots after color development of sulfuric acid ethanol solution and the colored spots, different spots represent different chemical components, not only the information spot complementation is exerted, but also the method is simple and convenient, the cost is low, the number of thin layer plates is small, the inspection condition is large, the detection information amount is large, the index is countable, and the novelty, the creativity and the practicability are self evident. At present, no report about a one-board multi-information thin-layer identification method of five-qi Korean-yang grass particles and standard decoction dry powder is found.
The technical scheme adopted by the invention for solving the technical problems is as follows:
grinding appropriate amount of five-qi Korean-yang grass granule and its standard decoction dry powder, respectively weighing 0.3g, adding 2ml methanol, ultrasonic treating for 10 min, centrifuging, and collecting supernatant as each sample solution; taking 1g of a five-gas Korean-herb control medicinal material, adding 50ml of 40% ethanol, performing reflux extraction for 30 minutes, filtering, concentrating the filtrate to dryness, adding 3ml of petroleum ether into residues, rotating a container to degrease, removing the petroleum ether, volatilizing until no ether smell exists, adding 1ml of 95% methanol into the residues, and dissolving to obtain a control medicinal material solution; sucking test solution 7 μ l and control solution 8-9 μ l, and respectively dropping on the same silica gel GF254Spreading toluene-ethyl acetate-formic acid-glacial acetic acid as a spreading agent in a volume ratio of 7: 6.3: 1.4: 0.2 on a thin layer plate, taking out, drying by hot air, placing in a dark room, inspecting by lamplight, and displaying main spots with the same color in the chromatogram of the test solution at the positions corresponding to the chromatogram of the reference solution; inspecting under 365nm ultraviolet lamp to obtain the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the reference materialA bright blue fluorescent main spot; inspecting under 254nm ultraviolet lamp to obtain the same brown main spot 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 to develop color, and inspecting under ultraviolet light 365nm to obtain main fluorescent spot with the same brightness, blue and green color in the sample chromatogram and the position corresponding to the control medicinal material chromatogram; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the position corresponding to the reference medicinal material chromatogram in the sample chromatogram.
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 one-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder. Namely, a test sample and a rubia yunnanensis medicinal material solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and after being unfolded, 17 information spots of five-qi cynomorium songaricum particles and standard decoction dry powder are detected under 5 detection conditions and belong to about 13 different chemical components. The spots before color development, the fluorescent spots, the non-fluorescent spots, the fluorescent spots after color development of sulfuric acid ethanol solution and the colored spots, different spots represent different chemical components, not only the information spot complementation is exerted, but also the method is simple and convenient, the cost is low, the number of thin layer plates is small, the inspection condition is large, the detection information amount is large, the index is countable, and the novelty, the creativity and the practicability are self evident. At present, no report on a one-board multi-information thin-layer identification method of five-qi Korean-yang grass particles and standard decoction dry powder is found.
2. The toluene-ethyl acetate-formic acid-glacial acetic acid with the volume ratio of 7: 6.3: 1.4: 0.2 is used as a developing agent, the five-qi chaoyang grass particles and the standard decoction dry powder are developed, after the thin-layer plate is dried by hot air, the thin-layer plate is inspected under sunlight, the medicinal materials, the particles and the standard decoction dry powder all present 1 main light grey spots, and the spots are inspected in a darkroom through lamplight, so that the definition of the spots is improved (figure 1); the medicinal materials and the samples are inspected under 365nm ultraviolet lamps, 5 fluorescent spots are inspected on the medicinal materials and the samples, 1 fluorescent spot on the medicinal materials is pink due to interference of chlorophyll, and the samples are all bright blue, so that the regulation is as follows: the same bright blue fluorescence main spot appears at the position corresponding to the color spectrum of the reference medicinal material, the color of the spot is limited, and the interference of the pink chlorophyll fluorescence spot is eliminated (figure 2); inspecting under 254nm ultraviolet lamp to obtain 4 brown spots with 1 high content (FIG. 3); spraying 10% sulfuric acid ethanol solution for color development, and detecting 3 fluorescent spots, wherein 2 fluorescent spots are newly added, one is bright blue, and the other is bright green (FIG. 4); the sample was placed in a dark room and the crude drug and sample were inspected for 4 tan spots by light (FIG. 5). Thus, on the same thin-layer plate, under five different inspection conditions, 17 information spots are detected in total. Comparing the spot size, shape, Rf values for the 5 inspection conditions, the spots were assigned to about 13 different chemical compositions.
3. The innovation point of the invention is that the components, polarity and separating capacity of the developing agent all achieve the expected target, so that the chemical components of the five-qi yang-ward herb are uniformly distributed in the whole area of the thin-layer plate, and the Rf values of the five-qi yang-ward herb all fall within the range of 0.2-0.8. Basically, a plurality of component spots are overlapped and crossed, but are complemented and interfered under respective inspection conditions, clear spots can be presented, information spot complementation is achieved, the purposes of minimum thin-layer plates, maximum inspection conditions and maximum detected information amount are achieved, and the technical difficulty is incomparable with that of conventional thin-layer identification. On the premise of not increasing any workload and detection cost, the detection index and supervision are greatly improved. 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 identification of the polar components in the various spot systems is detected.
5. Due to the particularity of chemical components contained in the five-qi yang-ward herb, in order to eliminate the interference of chlorophyll components, the water decoction and the decoction liquid which are the same as the granules are adopted to be concentrated to be dry, and a methanol constant volume mode is adopted, so that even if the decoction is carried out for 40 minutes and the concentration is carried out for 30 minutes, the information spots which are the same as the granules cannot be obtained, and the consideration is probably caused by the particularity of the contained chemical components. The requirements for thin layer identification are: the spots of the reference medicinal materials are almost the same as those of the samples, and the treatment method is as simple and rapid as possible, so that the method is a production process which can not be carried out for hours. Therefore, through the research and study of various treatment methods, 40% ethanol is used for reflux extraction, the extracting solution is volatilized, petroleum ether is degreased, 95% methanol is used for constant volume, only about 1 hour is needed, and the thin-layer result with basically consistent information spots of the reference medicinal material and the sample is obtained.
6. Compared with the method loaded by Chinese pharmacopoeia, the method has the advantages that the preparation time of the test solution is short, the solvent dosage is less, and no toxic solvent is concentrated and evaporated to dryness, and the most important is that: the gallic acid with poor specificity is not used as an index, but a plurality of chemical components are used as indexes, so that the detection information amount is greatly improved, and the method is simpler, quicker and more environment-friendly than the method of pharmacopoeia.
Drawings
FIG. 1 is a thin-layer TLC chart of five-qi Korean weed granules and standard decoction powder directly in a dark room after being developed and inspected by lamp light.
FIG. 2 is a TLC image of five-qi Korean weed granules and standard decoction powder, which are inspected under an ultraviolet lamp 365 nm.
FIG. 3 is a TLC image of five-qi Korean weed granules and standard decoction powder, which are inspected under an ultraviolet lamp 254 nm.
FIG. 4 is a TLC image of a thin layer of five-qi Korean weed granules and standard decoction dry powder, which are developed by spraying sulfuric acid ethanol solution and inspected under an ultraviolet lamp at 365 nm.
FIG. 5 is a TLC image of a thin layer of light inspection in a dark room after developing the five-qi Korean weed granules and the target decoction dry powder, spraying sulfuric acid ethanol solution for color development.
FIGS. 1, 2, 3, 4 and 5 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1.2 is Korean weed granules; 3. 4 is the five-qi Korean-yang grass decoction dry powder; 5.6. is prepared from radix seu herba Gei aleppici;
detailed description of the invention
Grinding appropriate amount of five-qi Korean-yang grass granule and its standard decoction dry powder, respectively weighing 0.3g, adding 2ml methanol, ultrasonic treating for 10 min, centrifuging, and collecting supernatant as each sample solution; taking 1g of a five-gas Korean-herb control medicinal material, adding 50ml of 40% ethanol, performing reflux extraction for 30 minutes, filtering, concentrating the filtrate to dryness, adding 3ml of petroleum ether into residues, rotating a container to degrease, removing the petroleum ether, volatilizing until no ether smell exists, adding 1ml of 95% methanol into the residues, and dissolving to obtain a control medicinal material solution; sucking test solution 7 μ l and control solution 8-9 μ l, and respectively dropping on the same silica gel GF254Spreading toluene-ethyl acetate-formic acid-glacial acetic acid as a spreading agent in a volume ratio of 7: 6.3: 1.4: 0.2 on a thin layer plate, taking out, drying by hot air, placing in a dark room, inspecting by lamplight, and displaying main spots with the same color in the chromatogram of the test solution at the positions corresponding to the chromatogram of the reference solution; inspecting under 365nm ultraviolet lamp to obtain bright blue fluorescent main spot with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the reference medicinal material; inspecting under 254nm ultraviolet lamp to obtain the same brown main spot 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 to develop color, and inspecting under ultraviolet light 365nm to obtain main fluorescent spot with the same brightness, blue and green color in the sample chromatogram and the position corresponding to the control medicinal material chromatogram; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the position corresponding to the reference medicinal material chromatogram in the sample chromatogram.
Claims (3)
1. A one-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder thereof is characterized in that:
grinding appropriate amount of five-qi Korean-yang grass granule and its standard decoction dry powder, respectively weighing 0.3g, adding 2ml methanol, ultrasonic treating for 10 min, centrifuging, and collecting supernatant as each sample solution; taking 1g of a five-gas Korean-herb control medicinal material, adding 50ml of 40% ethanol, performing reflux extraction for 30 minutes, filtering, concentrating the filtrate to dryness, adding 3ml of petroleum ether into residues, rotating a container to degrease, removing the petroleum ether, volatilizing until no ether smell exists, adding 1ml of 95% methanol into the residues, and dissolving to obtain a control medicinal material solution; sucking test solution 7 μ l and control solution 8-9 μ l, and respectively dropping on the same silica gel GF254Spreading toluene-ethyl acetate-formic acid-glacial acetic acid as a spreading agent in a volume ratio of 7: 6.3: 1.4: 0.2 on a thin layer plate, taking out, drying by hot air, placing in a dark room, inspecting by lamplight, and displaying main spots with the same color in the chromatogram of the test solution at the positions corresponding to the chromatogram of the reference solution; inspecting under 365nm ultraviolet lamp to obtain bright blue fluorescent main spot with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the reference medicinal material; inspecting under 254nm ultraviolet lamp to obtain the same brown main spot 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 to develop color, and inspecting under ultraviolet light 365nm to obtain main fluorescent spot with the same brightness, blue and green color in the sample chromatogram and the position corresponding to the control medicinal material chromatogram; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the position corresponding to the reference medicinal material chromatogram in the sample chromatogram.
2. The one-plate multi-information thin-layer identification method for the wuqi chaoyang grass granules and the standard decoction dry powder thereof according to claim 1, characterized in that each 1g of the granules is equivalent to 5g of crude drug; each 1g of standard decoction is equivalent to 5g of crude drug.
3. The method for identifying the one-plate multi-information thin layer of the five-qi korean sundew herb granules and the standard decoction dry powder thereof as claimed in claim 1, wherein the same developing agent is adopted, and 17 information spots of the five-qi korean sundew herb granules and the standard decoction dry powder thereof are detected on the same thin layer plate under 5 different detection conditions, and the 17 information spots belong to 13 different chemical components.
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| CN202111114093.7A CN113791166A (en) | 2021-09-23 | 2021-09-23 | One-plate multi-information thin-layer identification method for five-qi Korean-yang grass particles and standard decoction dry powder thereof |
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