CN111024879A - Rapid multi-information thin-layer identification method for fresh motherwort herb freeze-dried powder, particles and standard decoction dry powder - Google Patents
Rapid multi-information thin-layer identification method for fresh motherwort herb freeze-dried powder, particles and standard decoction dry powder Download PDFInfo
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Abstract
The invention relates to a rapid multi-information thin-layer identification method for fresh motherwort herb freeze-dried powder, granules and standard decoction dry powder. The method is characterized in that: a test sample and a fresh motherwort herb solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and after the development, 10 information spot components of the fresh motherwort herb freeze-dried powder, particles and standard decoction dry powder are inspected under 3 different inspection conditions, wherein the information spot components are fluorescent, non-fluorescent and colored when encountering an alkaloid reagent, are staggered but not mutually interfered, and the information spot complementation under different inspection conditions is exerted. Compared with the conventional alkaloid single information thin layer, the quality controllability of the identified object is greatly improved, the quality supervision is facilitated, and the method is simple, convenient and rapid, high in efficiency, low in cost and free of environmental pollution.
Description
Technical Field
The invention relates to a rapid multi-information thin-layer identification method for fresh motherwort herb freeze-dried powder, granules and standard decoction dry powder. Belongs to the field of thin-layer identification of traditional Chinese medicines. Namely, thin-layer chromatography is adopted for identification, and 10 information spots of the fresh motherwort herb freeze-dried powder, the particles and the standard decoction dry powder are detected on the same thin-layer plate under 3 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 are adopted, after ultrasonic extraction is carried out by using a proper solvent, filtration is not needed, the sample application, development and 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 effectiveness of the medication of people are ensured.
The Chinese medicinal motherwort is a variety collected in the calendar edition of Chinese pharmacopoeia and is processed according to the processing specifications of medicinal materials. Has the effects of promoting blood circulation, regulating menstruation, inducing diuresis, relieving swelling, and clearing away heat and toxic materials. Can be used for treating menoxenia, dysmenorrhea, amenorrhea, lochiorrhea, edema, oliguria, pyocutaneous disease, and toxic swelling. Is a commonly used gynecological traditional Chinese medicine in clinic. The main chemical components of the composition are alkaloids, flavonoids, diterpenes, phenylpropanol peptides, fatty acids, volatile oils, cyclic polypeptides and the like. The thin layer thereof identifies one part of the Chinese pharmacopoeia 2015 year edition【1】The stachydrine hydrochloride is adopted as a contrast, acetone-absolute ethyl alcohol-hydrochloric acid (10: 6: 1) is adopted as a developing agent, after development and air drying, the mixture is heated for 15 minutes at 105 ℃, cooled, sprayed with a mixed solution of diluted bismuth potassium iodide test solution-ferric trichloride test solution (10: 1) until the spots are clearly developed, and the color spots of the stachydrine hydrochloride are observed.
The literature reports that alkaloids are detected【2】However, 3g of motherwort herb control medicine is needed, 30ml of ethanol is firstly used for ultrasonic treatment for 30 minutes, the ethanol extract is concentrated to 5ml, the concentrated solution passes through a neutral alumina column and is then eluted by 30ml of ethanol, the eluent is evaporated to dryness, and 0.5ml of ethanol is added into the residue to be dissolved to be used as the control medicine solution.
At present, reports of other components detected by motherwort are not found, and reports of 10 information spots for simultaneously detecting other components, biological alkalinity and the like on the same thin-layer plate are not found.
The fresh motherwort freeze-dried powder is a processed product obtained by freeze drying fresh motherwort without sunshine or oven drying. Theoretically, the motherwort contains more effective components than the motherwort dried by the traditional processing procedure without any temperature; the particles and the standard powder are directly extracted from fresh motherwort, which is beneficial to the reservation of various effective components, and no report about the thin-layer identification of the fresh motherwort medicinal material is found at present, and no report about the rapid multi-information thin-layer identification method of the fresh motherwort medicinal material, the particles and the standard decoction dry powder is found.
The method provides scientific basis for the innovative preparation process of the fresh motherwort herb freeze-dried powder, the fresh motherwort herb granules and the fresh motherwort herb decoction dry powder for quick multi-information control, and carries out special thin-layer identification research on the fresh motherwort herb freeze-dried powder, the fresh motherwort herb granules and the fresh motherwort herb decoction dry powder with the purposes of simplicity, convenience, rapidness, low cost, high efficiency and multi-information. A set of simple, convenient and quick multi-information thin-layer identification method for fresh motherwort herb freeze-dried powder, granules and standard decoction dry powder without environmental pollution is obtained. The following processes for preparing lyophilized powder of fresh herba Leonuri, fresh herba Leonuri granule, and fresh herba Leonuri decoction are described as follows:
the fresh motherwort herb freeze-dried powder is prepared by removing impurities from a newly collected fresh motherwort herb, quickly cleaning, cutting, freeze-drying and crushing the fresh motherwort herb to obtain the fresh motherwort herb freeze-dried powder.
The fresh motherwort granule is prepared by removing impurities from fresh motherwort, quickly cleaning, directly decocting with water for 2 times, 1-2 hours each time, filtering, mixing filtrates, concentrating under reduced pressure to appropriate relative density, spray drying, adding appropriate amount of dextrin into dried powder, mixing well, and granulating.
The dry powder of the fresh motherwort target decoction is prepared by removing impurities from a newly collected fresh motherwort medicinal material, quickly cleaning, directly decocting with water for 2 times, 1-2 hours each time, filtering, combining filtrates, concentrating to a proper relative density at normal temperature, placing in a 60 ℃ oven, drying, and pulverizing.
Disclosure of Invention
Firstly creates a rapid multi-information thin-layer identification method for fresh motherwort herb freeze-dried powder, granules and standard decoction dry powder. Namely, a test sample and a fresh motherwort herb solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and after the development, 10 information spots of the fresh motherwort herb freeze-dried powder, the particles and the standard decoction dry powder are inspected under 3 different inspection conditions, wherein the information spots are fluorescent, non-fluorescent and colored when encountering an alkaloid reagent, are staggered but not mutually interfered, and the information spot complementation under different inspection conditions is exerted. Compared with the conventional alkaloid single information thin layer, the quality controllability of the identified object is greatly improved, the quality supervision is facilitated, and the method is simple, convenient and rapid, high in efficiency, low in cost and free of environmental pollution.
The technical scheme adopted by the invention for solving the technical problems is as follows:
taking 0.5g of each of fresh motherwort particles and fresh motherwort target decoction dry powder, adding 3ml of methanol respectively, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.3g of fresh motherwort herb freeze-dried powder, adding 2ml of methanol and 2 drops of dilute hydrochloric acid, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernatant as a medicinal solution; sucking 4 μ l of the test solution, 4 μ l of the dry powder of the standard decoction, and 10 μ l of the medicinal solution, and respectively dropping on the same silica gel GF254Spreading ethyl acetate-butanone-formic acid-water as a developing agent at a volume ratio of 4.5: 3: 1 on the thin layer plate, taking out, drying with hot air, inspecting under 365nm ultraviolet lamp, and displaying the same bright blue fluorescent spots in the sample chromatogram at the positions corresponding to the fresh herba Leonuri medicinal material lyophilized powder chromatogram; inspecting under 254nm ultraviolet lamp to obtain the same brown main spot in the sample chromatogram at the position corresponding to the chromatogram of fresh herba Leonuri lyophilized powder chromatogram; spraying a mixed solution of bismuth potassium iodide test solution and 10% sulphuric acid ethanol solution at a volume ratio of 10: 1, and observing in sunlight to obtain orange red main spots in the chromatogram of the test solution at the positions corresponding to the chromatogram of the lyophilized powder of fresh herba Leonuri.
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 fresh motherwort herb freeze-dried powder, granules and standard decoction dry powder. Namely, a test sample and a fresh motherwort herb solution are obtained by a simple and quick pretreatment method, and are respectively spotted on the same thin-layer plate, after the development, 10 information spot components of the fresh motherwort herb freeze-dried powder, the particles and the standard decoction dry powder are inspected under 3 different inspection conditions, wherein the components are fluorescent, non-fluorescent and colored when encountering an alkaloid reagent, and are staggered but not interfered with each other, so that the information spot complementation under different inspection conditions is exerted. Compared with the conventional alkaloid single information thin layer, the quality controllability of the identified object is greatly improved, the quality supervision is facilitated, and the method is simple, convenient and rapid, high in efficiency, low in cost and free of environmental pollution.
2. Using ethyl acetate-butanone-formic acid-water with a volume ratio of 4.5: 3: 1 as a developing agent, developing freeze-dried powder, particles and standard decoction dry powder of fresh motherwort herb, placing under an ultraviolet lamp of 365nm, and detecting 3 bright blue fluorescent spots of the herb, wherein the fluorescent spots of the test sample are redundant of the rest of the herb, which shows that the components change in the decocting and concentrating processes (figure 1); under 254nm ultraviolet lamp, 5 brown spots were detected on the medicinal materials, and the spots of the sample were also surplus medicinal materials, further proving that the components changed during the decoction and concentration process (fig. 2); from the Rf value analysis of fig. 1 and 2, the fluorescent spot and the non-fluorescent spot, which are not in the same position, indicate that they are not the same compound. After the spray alkaloid reagent is developed, clear alkaloid spots are presented in the medicinal material and the test solution, although the Rf value is low, no other spot interference exists, and the clear alkaloid spots can be used as evaluation indexes (figure 3); thus, the same thin-layer plate can be inspected for 10 information spot components under different inspection conditions. Innovates the pattern that motherwort is used for detecting alkaloid at present.
3. According to the identification method, the sample and the medicinal material solution can be obtained only by 0.3g of fresh motherwort medicinal material freeze-dried powder, 0.5g of particles and standard decoction dry powder respectively and 8ml of methanol within 20 minutes, and the developing time is 30 minutes plus 12ml of developing agent, and the total time is 1 hour, so that the multi-index evaluation can be performed on the sample quality, and the method is simple, convenient and quick and has high practical value.
4. The novel method is characterized in that 2ml of methanol and 2 drops of dilute hydrochloric acid are added into the medicinal material freeze-dried powder, the alkaloid is salified, the solubility is increased, the extraction amount of the alkaloid is greatly increased, the environmental pollution caused by adopting a concentration procedure due to low content at present is avoided, the detection sensitivity is improved, the use amount of the identified medicinal material is only 1/3-1/20 reported in documents, and the spots of the alkaloid are obvious and easy to judge.
5. After the thin layer of motherwort is developed, spraying bismuth potassium iodide test solution, so that spots sometimes do not appear, so that more help-seeking posts are on the net【3】. The identification has no spot after spraying the bismuth potassium iodide test solution, but 1ml of 10 percent ethanol sulfate solution is added into 10ml of the bismuth potassium iodide test solution, so that the problem that orange red spots of the alkaloid cannot be shown is solved, and the sensitivity of alkaloid color development is effectively improved.
Drawings
FIG. 1 is a thin-layer TLC chart of fresh herba Leonuri medicinal material, granule and target decoction dry powder under UV lamp 365 nm.
FIG. 2 is a thin-layer TLC chart of fresh herba Leonuri medicinal material, granule and target decoction dry powder under UV lamp 254 nm.
FIG. 3 is a TLC image of a thin layer under sunlight after the color development of fresh herba Leonuri medicinal material, particles and target decoction dry powder with alkaloid color developing agent.
FIGS. 1, 2 and 3 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1.2.3 is Xianyi
Freeze-dried powder of a mother herbal medicine; 4 is dry powder of decoction of fresh motherwort herb; 5.6 is fresh motherwort grains;
the specific implementation mode of the invention is as follows:
taking 0.5g of each of fresh motherwort particles and fresh motherwort target decoction dry powder, adding 3ml of methanol respectively, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.3g of fresh motherwort herb freeze-dried powder, adding 2ml of methanol and 1 drop of dilute hydrochloric acid, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernatant as a medicinal solution; sucking 4 μ l of the test solution, 4 μ l of the dry powder of the standard decoction, and 10 μ l of the medicinal solution, and respectively dropping on the same silica gel GF254Spreading ethyl acetate-butanone-formic acid-water as a developing agent at a volume ratio of 4.5: 3: 1 on the thin layer plate, taking out, drying with hot air, inspecting under 365nm ultraviolet lamp, and displaying the same bright blue fluorescent spots in the sample chromatogram at the positions corresponding to the fresh herba Leonuri medicinal material lyophilized powder chromatogram; inspecting under 254nm ultraviolet lamp to obtain the same brown main spot in the sample chromatogram at the position corresponding to the chromatogram of fresh herba Leonuri lyophilized powder chromatogram; spraying a mixed solution of a bismuth potassium iodide test solution and a 10% sulfuric acid ethanol solution in a volume ratio of 10: 1, wherein the same orange red main spots appear in the chromatogram of the test solution at the positions corresponding to the chromatogram of the freeze-dried powder of the fresh motherwort herb.
Reference to the literature
【1】 Pharmacopoeia committee of the people' S republic of china pharmacopoeia 2015 year edition (part) [ S ]. beijing: chinese medical science and technology press, month 2015.6: 290 page 291
【2】 Liao Wei, Wang Xiaoying, thin layer identification of Compound motherwort granule, today's pharmacy, 2009,19(2):44-45, page 35
【3】 Dandelion forum-pharmaceutical quality-quality control-help: the thin layer of motherwort was identified as showing no corresponding spots and was published in 2016-6-1410: 40: 34.
Claims (3)
1. A rapid multi-information thin-layer identification method for fresh motherwort medicinal materials, particles and standard decoction dry powder is characterized in that:
taking 0.5g of each of fresh motherwort particles and fresh motherwort target decoction dry powder, adding 3ml of methanol respectively, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.3g of fresh motherwort herb freeze-dried powder, adding 2ml of methanol and 2 drops of dilute hydrochloric acid, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernatant as a medicinal solution; sucking 4 μ l of the test solution, 4 μ l of the dry powder of the standard decoction, and 10 μ l of the medicinal solution, and respectively dropping on the same silica gel GF254Spreading ethyl acetate-butanone-formic acid-water as a developing agent at a volume ratio of 4.5: 3: 1 on the thin layer plate, taking out, drying with hot air, inspecting under 365nm ultraviolet lamp, and displaying the same bright blue fluorescent spots in the sample chromatogram at the positions corresponding to the fresh herba Leonuri medicinal material lyophilized powder chromatogram; inspecting under 254nm ultraviolet lamp to obtain the same brown main spot in the sample chromatogram at the position corresponding to the chromatogram of fresh herba Leonuri lyophilized powder chromatogram; spraying a mixed solution of bismuth potassium iodide test solution and 10% sulphuric acid ethanol solution at a volume ratio of 10: 1, and observing in sunlight to obtain orange red main spots in the chromatogram of the test solution at the positions corresponding to the chromatogram of the lyophilized powder of fresh herba Leonuri.
2. The method of claim 1, wherein the lyophilized powder of herba Leonuri is equivalent to fresh herba Leonuri 15g per 1 g; each 1g of the granules is 15g of fresh medicinal materials; each 1g of standard decoction is 15g of fresh medicinal materials.
3. The method of claim 1, wherein 10 information spots of the fresh motherwort herb, the particles and the standard decoction dry powder are inspected on the same developing agent, the same thin layer plate and 3 different inspection conditions.
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