CN111458450B - Rapid multi-information thin-layer identification method for sticktight particles and decocting powder - Google Patents
Rapid multi-information thin-layer identification method for sticktight particles and decocting powder Download PDFInfo
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- CN111458450B CN111458450B CN202010279160.XA CN202010279160A CN111458450B CN 111458450 B CN111458450 B CN 111458450B CN 202010279160 A CN202010279160 A CN 202010279160A CN 111458450 B CN111458450 B CN 111458450B
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Abstract
The invention relates to a quick multi-information thin-layer identification method for sticktight particles and standard decoction powder. The method is characterized in that: a test sample and a medicinal material solution are obtained by a simple and quick pretreatment method, and are respectively spotted on the same thin-layer plate, and 6 information spots of the medicinal materials, the particles and the standard decoction powder of the bidens bipinnata are inspected under 4 different inspection conditions after the spreading. The fluorescence spot and the color spot are mutually staggered but not interfered when meeting 10 percent ethanol sulfate solution, and the information spot complementation under different inspection conditions is exerted. Compared with the conventional thin-layer identification, the quality controllability of the identified object is greatly improved, the quality supervision is facilitated, and the method is simple, convenient and quick, has high efficiency and low cost and does not pollute the environment.
Description
Technical Field
The invention relates to a quick multi-information thin-layer identification method for sticktight particles and standard decoction powder. Belongs to the field of thin-layer identification of traditional Chinese medicines. Namely, thin-layer chromatography is adopted for identification, and 6 information spots of the Bidens pilosa medicinal material, the grains and the standard decoction powder are detected on the same thin-layer plate under 4 different detection conditions.
Background
The so-called rapid multi-information thin-layer identification method is simple, convenient, rapid and efficient as the name implies, the sample solution and the reference medicinal material solution do not need repeated purification treatment, no operation procedures such as evaporation, concentration and the like polluting the environment, after ultrasonic treatment by using a proper solvent, filtration is not needed, the sample application, the development and the inspection of the supernatant are carried out, and the thin-layer result can be obtained quickly. The multi-information refers to a thin layer plate which is unfolded for 1 time, and various information spots under different inspection conditions can be obtained. Compared with the conventional thin-layer identification, the quality controllability of the detected object is greatly improved, the market quality supervision is facilitated, and the safety and the effectiveness of the drug use of people are ensured.
Bidens pilosa is the whole plant of Bidens pilosa L.of Compositae, is a common Chinese herbal medicine, has effects of clearing heat and detoxicating, removing blood stasis and promoting blood circulation, and is mainly used for treating upper respiratory infection, sore throat, acute appendicitis, acute icteric hepatitis, gastroenteritis, rheumatic arthralgia and malaria, and for treating sore and furuncle, venomous snake bite and traumatic injury. Mainly contains flavonoids such as hyperoside, ocannin, echinacoside, quercetin and glycosides formed by the quercetin and the saccharides, but the Chinese pharmacopoeia is not yet introduced, so the legal quality standard of the Chinese pharmacopoeia is not reported. Only consult one【1】A Master academic paper of Bidens pilosa medicinal material quality standard research in Shanxi province describes a thin-layer identification method of Bidens pilosa, and a preparation method of a test solution comprises the following steps: collecting herba Bidentis Bipinnatae powder 5.0g, adding methanol 50ml, heating under reflux for 2 hr, filtering, evaporating filtrate to dryness, dissolving residue with ethyl acetate 10ml, extracting with distilled water for 2 times, each time 10ml, mixing water layers, evaporating to dryness, and dissolving residue with methanol 2ml to obtain sample solution. Taking appropriate amount of rutin and hyperoside, adding methanol respectively to obtain solution containing rutin and hyperoside 0.2mg each per 1ml, and using as control solution. Sucking sample solution and control solution 5 μ l each, dropping on the same silica gel G thin layer plate, developing with ethyl acetate-butanone-formic acid-water (5: 3: 1) as developing agent, taking out, air drying, spraying with aluminum trichloride ethanol solution, drying, placing under ultraviolet lamp 365nm, and inspecting spot color. The method has large sampling amount of herba Bidentis Bipinnatae (5.0 g medicinal material); the preparation of the test solution takes a long time, at least 4 hours; the reference substance is used as a reference, the spots are single, the detection indexes are few, and the quality supervision of multiple indexes is not facilitated. The thin-layer identification of the prepared granules and the standard decoction powder and the rapid multi-information thin-layer identification method of the medicinal materials, the granules and the standard decoction powder of the bidens pilosa are not referred to in literature reports.
The method provides scientific basis for the preparation process of the bidens bipinnata particles and the standard decoction powder for quick multi-information control, and performs special thin-layer identification research on the bidens bipinnata particles and the standard decoction powder according to the purposes of simplicity, rapidness, low cost, high efficiency and multi-information. A set of simple, convenient and quick thin-layer identification method for sticktight particles and standard decoction powder without environmental pollution is obtained. The following process for preparing Bidens pilosa medicinal materials, granules and decoction powder is described as follows:
the herba Bidentis Bipinnatae medicinal material is prepared by collecting herba Bidentis Bipinnatae, removing impurities, rapidly cleaning, drying, and cutting.
The herba Bidentis Bipinnatae granule is prepared by decocting herba Bidentis Bipinnatae with water for 2 times, each time for 1-2 hr, filtering, mixing filtrates, concentrating under reduced pressure to appropriate relative density, spray drying, adding appropriate amount of dextrin into dried powder, mixing, and making into granule.
The marked herba Bidentis Bipinnatae decoction powder is prepared by decocting herba Bidentis Bipinnatae with water for 2 times, each for 1 hr, filtering, mixing filtrates, concentrating at room temperature to appropriate relative density, oven drying at 60 deg.C, and pulverizing.
Disclosure of Invention
Firstly creates a rapid multi-information thin-layer identification method for sticktight particles and standard decoction powder. Namely: a test sample and a medicinal material solution are obtained by a simple and quick pretreatment method, and are respectively spotted on the same thin-layer plate, and 6 information spots of the medicinal materials, the particles and the standard decoction powder of the bidens bipinnata are inspected under 4 different inspection conditions after the spreading. The fluorescence spot and the color spot are mutually staggered but not interfered when meeting 10 percent ethanol sulfate solution, and the information spot complementation under different inspection conditions is exerted. Compared with the conventional thin-layer identification, the quality controllability of the identified object is greatly improved, the quality supervision is facilitated, and the method is simple, convenient and quick, has high efficiency and low cost and does not pollute the environment.
The technical scheme adopted by the invention for solving the technical problems is as follows:
collecting herba Bidentis Bipinnatae granule and herba Bidentis Bipinnatae decoction powder 0.5g, adding methanol 3ml, respectively, ultrasonic treating for 10 min, centrifuging, and collecting supernatantA test solution; taking 0.2g of sticktight medicinal material powder, adding 2ml of methanol, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernate as a medicinal material solution; sucking 4 μ l of granule sample solution, 6 μ l of standard decoction powder sample solution, and 6 μ l of medicinal material solution, respectively dropping on the same silica gel GF254Spreading on a thin layer plate with ethyl acetate-methanol-formic acid-water as developing agent at volume ratio of 5: 2: 0.2, taking out, drying with hot air, inspecting under ultraviolet lamp 365nm, and displaying the same bright blue fluorescence main spot in the sample chromatogram at the position corresponding to the herba Bidentis Bipinnatae medicinal material chromatogram; inspecting under 254nm ultraviolet lamp to obtain main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of herba Bidentis Bipinnatae; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color clearly, and inspecting under ultraviolet lamp 365nm to obtain blue-green fluorescence main spot at the position corresponding to herba Bidentis Bipinnatae medicinal material chromatogram in the sample chromatogram; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the position corresponding to the color spectrum of herba Bidentis Bipinnatae in the color spectrum of the sample.
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. And then, by means of effective components with similar polarities, the effective components are overlapped on the same thin-layer plate under different inspection conditions, but are not interfered with each other on different layers, so that different color spots are displayed, and the 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 sticktight particles and standard decoction powder. Namely: a test sample and a medicinal material solution are obtained by a simple and quick pretreatment method, and are respectively spotted on the same thin-layer plate, and 6 information spots of the medicinal materials, the particles and the standard decoction powder of the bidens bipinnata are inspected under 4 different inspection conditions after the spreading. The fluorescence spot and the color spot are mutually staggered but not interfered when meeting 10 percent ethanol sulfate solution, and the information spot complementation under different inspection conditions is exerted. Compared with the conventional thin-layer identification, the quality controllability of the identified object is greatly improved, the quality supervision is facilitated, and the method is simple, convenient and quick, has high efficiency and low cost and does not pollute the environment.
2. Spreading herba Bidentis Bipinnatae medicinal materials, granules and decoction powder with ethyl acetate-methanol-formic acid-water at volume ratio of 5: 2: 0.2 as developing agent, and inspecting under ultraviolet lamp 365nm to obtain 1 bright blue fluorescent spot (figure 1); inspecting under 254nm ultraviolet lamp to obtain 2 brown spots, 3-4 brown spots and 5-6 brown spots on herba Bidentis Bipinnatae decoction powder, so as to reveal that the chemical components of spray-dried granules are more than those of normal-temperature concentrated standard decoction powder, and the heated and extracted granules and standard decoction powder are more than those of unextracted medicinal materials; comparing the shape and size of the spots in FIGS. 1 and 2, the fluorescent spot and the tan spot are not the same compound. Spraying 10% sulphuric acid ethanol solution for color development, and inspecting under 365nm ultraviolet lamp to obtain 1 clear blue-green fluorescence main spot (figure 3) on medicinal material and sample, wherein the spot intensity and size of all the detected substances are the same, and the spot has different Rf value from the corresponding spot in figures 1 and 2, is not the same compound, and is a new component spot excited after spraying sulphuric acid ethanol solution for color development; viewing in dark room, viewing 2 red brown spots on the medicinal materials and the sample, comparing with the spots in figures 1, 2, and 3, and showing new components after coloring with sulfuric acid, wherein Rf value, shape, dosage, and other spots are different. Thus, under 4 different inspection conditions, a total of 6 information spots, i.e., 3 spots before development and 3 spots after development, were detected. They are mutually staggered, but do not interfere with each other under respective inspection conditions, and play a role in spot complementation under different inspection conditions.
3. Although the identified Bidens pilosa medicinal material belongs to the whole grass class, contains a large amount of chlorophyll, and is red fluorescent spot under the ultraviolet lamp of 365nm, which interferes description. However, the adopted developing agent pushes the chlorophyll red spots to the forefront of the thin-layer plate (figure 1) to exceed the specified Rf value range of 0.2-0.8, and the defined bright blue fluorescence main spots are added, so that the red chlorophyll fluorescence spots do not interfere with the result judgment any more. The purification treatment for removing chlorophyll is omitted, and the pretreatment time of the reference medicinal materials is saved.
4. Under 4 inspection conditions, the spots of fat-soluble components are shown in fig. 1 and 2, the spots of fat-soluble components are shown in fig. 3 and 4, the components with higher water solubility are shown, the content of the components is higher than that of the fat-soluble components, the treatment effect of the traditional water decoction is reflected, and the effective components are mostly water-soluble.
5. The identification method can obtain a sample and a medicinal material solution only by 0.2g of the sticktight medicinal material, 0.5g of the particles and the standard decoction powder respectively and 8ml of methanol within 20 minutes, and can evaluate the quality of the sticktight particles and the standard decoction powder by adding 12ml of the developing agent, wherein the developing time is 30 minutes and takes 1 hour totally, thus being simple, convenient and quick and having higher practical value.
Drawings
FIG. 1 is a TLC image of herba Bidentis Bipinnatae medicinal materials, granules and decoction powder under 365nm UV light.
FIG. 2 is a TLC image of Bidens pilosa herbs, granules and decocted powder under 254nm UV light.
FIG. 3 is a TLC chart of the thin layer of the Bidens pilosa herb, granule and decoction powder after being developed with 10% ethanol sulfate solution and being inspected under the ultraviolet lamp 365 nm.
FIG. 4 is a thin-layer TLC chart of the color development of Bidens pilosa herb, granule and decoction powder in 10% ethanol sulfate solution and the inspection by lamp light in a dark room.
FIGS. 1, 2, 3 and 4 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1.2.3 is Bidens pilosa medicinal material; 4. is herba Bidentis Bipinnatae decoction powder; 5.6. is herba Bidentis Bipinnatae granule.
The specific implementation mode of the invention is as follows:
collecting ramulus Et folium Bidentis BipinnataeRespectively adding 0.5g of grass particles and 0.5g of sticktight decoction powder into 3ml of methanol, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.2g of sticktight medicinal material powder, adding 2ml of methanol, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernate as a medicinal material solution; sucking 4 μ l of granule sample solution, 6 μ l of standard decoction powder sample solution, and 6 μ l of medicinal material solution, respectively dropping on the same silica gel GF254Spreading on a thin layer plate with ethyl acetate-methanol-formic acid-water as developing agent at volume ratio of 5: 2: 0.2, taking out, drying with hot air, inspecting under ultraviolet lamp 365nm, and displaying the same bright blue fluorescence main spot in the sample chromatogram at the position corresponding to the herba Bidentis Bipinnatae medicinal material chromatogram; inspecting under 254nm ultraviolet lamp to obtain main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of herba Bidentis Bipinnatae; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color clearly, and inspecting under ultraviolet lamp 365nm to obtain blue-green fluorescence main spot at the position corresponding to herba Bidentis Bipinnatae medicinal material chromatogram in the sample chromatogram; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the position corresponding to the color spectrum of herba Bidentis Bipinnatae in the color spectrum of the sample.
Reference to the literature
【1】 The research on the quality standard of Bidens pilosa in Shanxi province, Master's academic thesis of Shanxi medical university, Buziyun, 2015.05.20.
Claims (3)
1. A quick multi-information thin-layer identification method for sticktight particles and standard decoction powder is characterized in that:
taking 0.5g of herba Bidentis Bipinnatae granule and herba Bidentis Bipinnatae decoction powder respectively, adding 3ml of methanol, performing ultrasonic treatment for 10 min, centrifuging, and collecting supernatant as sample solution; taking 0.2g of sticktight medicinal material powder, adding 2ml of methanol, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernate as a medicinal material solution; sucking 4 μ l of granule sample solution, 6 μ l of standard decoction powder sample solution, and 6 μ l of medicinal material solution, respectively dropping on the same silica gel GF254Spreading with ethyl acetate-methanol-formic acid-water at volume ratio of 5: 2: 0.2 as developing agent, taking out, drying with hot air, inspecting under ultraviolet lamp 365nm to obtain test sampleIn the chromatogram, the same bright blue fluorescence main spot appears at the position corresponding to the medicinal material chromatogram of the bidens bipinnata; inspecting under 254nm ultraviolet lamp to obtain main spots with the same color in the chromatogram of the sample at the position corresponding to the chromatogram of herba Bidentis Bipinnatae; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color clearly, and inspecting under ultraviolet lamp 365nm to obtain blue-green fluorescence main spot at the position corresponding to herba Bidentis Bipinnatae medicinal material chromatogram in the sample chromatogram; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the position corresponding to the color spectrum of herba Bidentis Bipinnatae in the color spectrum of the sample.
2. The method for rapid multi-information thin-layer identification of sticktight particles and decoction powders according to claim 1, wherein each 1g of particles is equivalent to 6g of crude drug; each 1g of decocted powder is equivalent to 6g of crude drug.
3. The method of claim 1, wherein the same developer is used to inspect 6 information spots of Bidens bipinnata herbs, Bidens bipinnata particles and Bidens bipinnata decoction powder under 4 different inspection conditions on the same thin layer plate.
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| CN101890066A (en) * | 2010-07-08 | 2010-11-24 | 深圳市药品检验所 | Method for constructing Bidens parviflor willd medicinal material capillary electrophoresis fingerprint and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101890066A (en) * | 2010-07-08 | 2010-11-24 | 深圳市药品检验所 | Method for constructing Bidens parviflor willd medicinal material capillary electrophoresis fingerprint and application thereof |
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