CN110824094A - Gradient full-information thin-layer identification method for eclipta medicinal materials - Google Patents
Gradient full-information thin-layer identification method for eclipta medicinal materials Download PDFInfo
- Publication number
- CN110824094A CN110824094A CN201911069548.0A CN201911069548A CN110824094A CN 110824094 A CN110824094 A CN 110824094A CN 201911069548 A CN201911069548 A CN 201911069548A CN 110824094 A CN110824094 A CN 110824094A
- Authority
- CN
- China
- Prior art keywords
- chromatogram
- medicinal material
- solution
- thin
- spots
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000463 material Substances 0.000 title claims abstract description 67
- 238000000034 method Methods 0.000 title claims abstract description 28
- 241001421953 Eclipta <beetle> Species 0.000 title claims 7
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 29
- 239000002904 solvent Substances 0.000 claims abstract description 8
- 238000000605 extraction Methods 0.000 claims abstract description 5
- 239000012085 test solution Substances 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 32
- 239000012088 reference solution Substances 0.000 claims description 20
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 13
- 238000007605 air drying Methods 0.000 claims description 13
- 239000000741 silica gel Substances 0.000 claims description 13
- 229910002027 silica gel Inorganic materials 0.000 claims description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 238000005507 spraying Methods 0.000 claims description 9
- PJWKAIDLIGVSCP-UHFFFAOYSA-N OC=O.CCOC(C)=O.C1CCCCC1 Chemical compound OC=O.CCOC(C)=O.C1CCCCC1 PJWKAIDLIGVSCP-UHFFFAOYSA-N 0.000 claims description 8
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 6
- NOTFZGFABLVTIG-UHFFFAOYSA-N Cyclohexylethyl acetate Chemical compound CC(=O)OCCC1CCCCC1 NOTFZGFABLVTIG-UHFFFAOYSA-N 0.000 claims description 4
- WIYUJTDPESFNJV-UHFFFAOYSA-N chloroform formic acid methanol hydrate Chemical compound O.OC.OC=O.ClC(Cl)Cl WIYUJTDPESFNJV-UHFFFAOYSA-N 0.000 claims description 4
- 238000009210 therapy by ultrasound Methods 0.000 claims description 4
- 238000000227 grinding Methods 0.000 claims description 3
- 244000286838 Eclipta prostrata Species 0.000 abstract description 33
- 238000007689 inspection Methods 0.000 abstract description 15
- 150000001875 compounds Chemical class 0.000 abstract description 8
- 238000002360 preparation method Methods 0.000 abstract description 6
- 239000012488 sample solution Substances 0.000 abstract description 6
- 238000012360 testing method Methods 0.000 abstract description 5
- 238000002203 pretreatment Methods 0.000 abstract description 2
- 238000011161 development Methods 0.000 description 9
- 239000000523 sample Substances 0.000 description 8
- 238000004809 thin layer chromatography Methods 0.000 description 8
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- KIWBPDUYBMNFTB-UHFFFAOYSA-N Ethyl hydrogen sulfate Chemical compound CCOS(O)(=O)=O KIWBPDUYBMNFTB-UHFFFAOYSA-N 0.000 description 5
- 241000446313 Lamella Species 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 239000003086 colorant Substances 0.000 description 4
- 238000003892 spreading Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 208000032843 Hemorrhage Diseases 0.000 description 2
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 description 2
- YTPLMLYBLZKORZ-UHFFFAOYSA-N Thiophene Chemical compound C=1C=CSC=1 YTPLMLYBLZKORZ-UHFFFAOYSA-N 0.000 description 2
- 208000031971 Yin Deficiency Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000003795 desorption Methods 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000013441 quality evaluation Methods 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 229930192474 thiophene Natural products 0.000 description 2
- OHZAHWOAMVVGEL-UHFFFAOYSA-N 2,2'-bithiophene Chemical compound C1=CSC(C=2SC=CC=2)=C1 OHZAHWOAMVVGEL-UHFFFAOYSA-N 0.000 description 1
- KXSFECAJUBPPFE-UHFFFAOYSA-N 2,2':5',2''-terthiophene Chemical compound C1=CSC(C=2SC(=CC=2)C=2SC=CC=2)=C1 KXSFECAJUBPPFE-UHFFFAOYSA-N 0.000 description 1
- SMRPGWBDLOQHOS-UHFFFAOYSA-N 5-[4,5-dihydroxy-6-(hydroxymethyl)-3-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxy-3,4-dihydroxy-6-[[9-hydroxy-4-(hydroxymethyl)-4,6a,6b,8a,11,11,14b-heptamethyl-14-oxo-2,3,4a,5,6,7,8,9,10,12,12a,14a-dodecahydro-1H-picen-3-yl]oxy]oxane-2-carboxylic acid Chemical compound OC1C(O)C(O)C(C)OC1OC1C(OC2C(OC(C(O)C2O)C(O)=O)OC2C(C3C(C4C(C5(CCC6(C)C(O)CC(C)(C)CC6C5=CC4=O)C)(C)CC3)(C)CC2)(C)CO)OC(CO)C(O)C1O SMRPGWBDLOQHOS-UHFFFAOYSA-N 0.000 description 1
- WYDPEADEZMZKNM-ZBKPBKBGSA-N Echinocystic acid 3-glucoside Chemical compound O([C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)C[C@@H](O)[C@]1(CCC(C[C@H]14)(C)C)C(O)=O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O WYDPEADEZMZKNM-ZBKPBKBGSA-N 0.000 description 1
- WYDPEADEZMZKNM-UHFFFAOYSA-N Ecliptasaponin D Natural products C12CC(C)(C)CCC2(C(O)=O)C(O)CC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OC(CO)C(O)C(O)C1O WYDPEADEZMZKNM-UHFFFAOYSA-N 0.000 description 1
- 208000034507 Haematemesis Diseases 0.000 description 1
- 206010027514 Metrorrhagia Diseases 0.000 description 1
- QIFYGFMZFHRIBM-UHFFFAOYSA-N O.CO.C(C)(=O)OCC.ClCCl Chemical compound O.CO.C(C)(=O)OCC.ClCCl QIFYGFMZFHRIBM-UHFFFAOYSA-N 0.000 description 1
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 description 1
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 description 1
- BMWPBKOFJSHJAW-UHFFFAOYSA-N Saponin B Natural products CC1(C)CCC2(CCC3(C)C(=CCC4C5(C)CCC(OC6OC(CO)C(O)C(OC7OC(CO)C(O)C(O)C7O)C6=O)C(C)(C)C5CCC34C)C2C1)C(=O)O BMWPBKOFJSHJAW-UHFFFAOYSA-N 0.000 description 1
- 208000009205 Tinnitus Diseases 0.000 description 1
- 206010053476 Traumatic haemorrhage Diseases 0.000 description 1
- 235000005811 Viola adunca Nutrition 0.000 description 1
- 240000009038 Viola odorata Species 0.000 description 1
- 235000013487 Viola odorata Nutrition 0.000 description 1
- 235000002254 Viola papilionacea Nutrition 0.000 description 1
- XADJWCRESPGUTB-UHFFFAOYSA-N apigenin Natural products C1=CC(O)=CC=C1C1=CC(=O)C2=CC(O)=C(O)C=C2O1 XADJWCRESPGUTB-UHFFFAOYSA-N 0.000 description 1
- KZNIFHPLKGYRTM-UHFFFAOYSA-N apigenin Chemical compound C1=CC(O)=CC=C1C1=CC(=O)C2=C(O)C=C(O)C=C2O1 KZNIFHPLKGYRTM-UHFFFAOYSA-N 0.000 description 1
- 235000008714 apigenin Nutrition 0.000 description 1
- 229940117893 apigenin Drugs 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 208000001848 dysentery Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 208000001780 epistaxis Diseases 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 208000006750 hematuria Diseases 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- KDCIHNCMPUBDKT-UHFFFAOYSA-N hexane;propan-2-one Chemical compound CC(C)=O.CCCCCC KDCIHNCMPUBDKT-UHFFFAOYSA-N 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 210000003127 knee Anatomy 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- IQPNAANSBPBGFQ-UHFFFAOYSA-N luteolin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(O)C(O)=C1 IQPNAANSBPBGFQ-UHFFFAOYSA-N 0.000 description 1
- LRDGATPGVJTWLJ-UHFFFAOYSA-N luteolin Natural products OC1=CC(O)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 LRDGATPGVJTWLJ-UHFFFAOYSA-N 0.000 description 1
- 235000009498 luteolin Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- LVTJOONKWUXEFR-UEZXSUPNSA-N protodioscin Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@H](O)[C@@H](O)[C@H](C)O1)O)O[C@@H]1CC2=CC[C@H]3[C@@H]4C[C@@H]5O[C@]([C@H]([C@@H]5[C@@]4(C)CC[C@@H]3[C@@]2(C)CC1)C)(O)CC[C@@H](C)CO[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O LVTJOONKWUXEFR-UEZXSUPNSA-N 0.000 description 1
- 229960001285 quercetin Drugs 0.000 description 1
- 235000005875 quercetin Nutrition 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 230000002787 reinforcement Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- PPRSVUXPYPBULA-UHFFFAOYSA-N saponin A Natural products CC1(C)CCC2(CCC3(C)C(=CCC4C5(C)CCC(OC6OC(CO)C(O)C(O)C6=O)C(C)(C)C5CCC34C)C2C1)C(=O)O PPRSVUXPYPBULA-UHFFFAOYSA-N 0.000 description 1
- QDQWGYLCDZBAMD-UHFFFAOYSA-N saponin C Natural products CC1C2C3CCC4C5(C)CCC(O)C(C)(COC6OC(CO)C(O)C(O)C6O)C5CCC4(C)C3(C)CCC27C8OC8C1(C)OC7=O QDQWGYLCDZBAMD-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 150000003577 thiophenes Chemical class 0.000 description 1
- 231100000886 tinnitus Toxicity 0.000 description 1
- 150000008130 triterpenoid saponins Chemical class 0.000 description 1
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 description 1
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 description 1
- 235000012141 vanillin Nutrition 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/94—Development
Abstract
The invention relates to a gradient full-information thin-layer identification method for an eclipta medicinal material. The method is characterized in that: a test sample solution and a reference medicinal material solution are obtained by a simple and quick pretreatment method, and about 27 information spots of the eclipta medicinal material are inspected on 4 thin-layer plates by adopting the same test sample solution and 4 developing agents with different polarities under various inspection conditions. The components are crossed with each other, but do not interfere with each other under the respective inspection conditions, and clear fluorescent spots or color spots can be displayed. The thin-layer gradient identification is completed by 0.2g of each of the medicinal material and the reference medicinal material, 4ml of extraction solvent, 40ml of developing solvent and 1.5 hours. Simple, convenient, quick and efficient, and is not available in the reported methods at present. Provides a method and a chromatogram map for multi-information quick quality supervision of the yerbadetajo herb; provides identification references of components with different polarities for the compound preparation.
Description
Technical Field
The invention relates to a gradient full-information thin-layer identification method for an eclipta medicinal material. Namely, all the information component spots from fat solubility to water solubility in the eclipta medicinal material are detected in a thin-layer identification mode according to the gradient sequence of the fat solubility, the medium polarity and the water solubility.
Background
The gradient complete information thin-layer identification of the traditional Chinese medicinal materials is characterized in that the content of high-performance liquid phase gradient elution is expanded to the field of thin-layer identification of the traditional Chinese medicinal materials, fat-soluble, medium-polar and water-soluble components in the traditional Chinese medicinal materials are repeatedly adsorbed and desorbed in developing solvents with different polarities according to similar compatible dissolving laws and in the sequence of non-polarity, medium-polarity and polarity, most of effective components with detection information in the medicinal materials are obtained on a few thin-layer plates according to the polarity size under different conditions to obtain a spot information map which is as complete as possible, so that a set of complete information thin-layer chromatogram and identification method from the fat-soluble to the water-soluble components is formed, and the eclipta medicinal materials from different sources are provided for integral quality supervision; and a compound preparation prepared by various processes as a quality evaluation reference map and a method.
The full information means that no information spots are present at the forefront of a lamella plate of a detection part of the fat-soluble component, no obvious spot residue is present at a base line of the lamella plate of the detection part of the water-soluble component, all information spots are included in a gradient lamella spreading range, and the information spots capable of being detected are all present under different spreading agents and detection conditions by matching with various detection conditions, so that the fat-soluble to water-soluble full information content is obtained.
The traditional Chinese medicine eclipta is a variety collected in the calendar edition of Chinese pharmacopoeia, has the effects of nourishing liver and kidney, cooling blood and stopping bleeding, and is used for liver and kidney yin deficiency, tooth loosening, early white beard and hair, dizziness and tinnitus, soreness and weakness of waist and knees, yin deficiency blood heat hematemesis, epistaxis, hematuria, bloody dysentery, metrorrhagia and metrostaxis and traumatic hemorrhage. Is a clinical tonicIt is combined with the commonly used herbs for stopping bleeding. Contains triterpenoid saponins, such as Ecliptae herba saponin A, Ecliptae herba saponin B, and Ecliptae herba saponin C; flavonoids such as apigenin, luteolin, quercetin and glycosides thereof; thiophenes, such as bithiophene, terthiophene, and the like. These components, from structural analysis, are both lipid-soluble, medium-polar and water-soluble. But the thin layer is used for identifying one part of the national pharmacopoeia 2015 edition, the adopted eclipta saponin A and a reference medicinal material are used as a reference, dichloromethane-ethyl acetate-methanol-water (30: 40: 15: 3) is used as a developing agent, and after the development, vanillin sulfuric acid test solution is sprayed for developing color, and the developed spots are detected. From the polarity analysis of the developer, the information spots examined were of a more water-soluble nature. There are reports in the literature【1】Taking 1g of Ecliptae herba powder, adding 20ml of ethanol, soaking for 2 hours, performing ultrasonic treatment for 30 minutes, filtering, volatilizing the filtrate, and adding 1ml of absolute ethanol to dissolve residues to obtain a test solution. Taking 1g of herba Ecliptae as control material, and making into control solution in the same way. Sucking the two solutions, respectively dropping the solutions in strips on high performance silica gel prefabricated thin layer plates (HPTLC-plate Nano-DURASIL-20, MN), taking n-hexane-acetone (9: 1) as developing agent, pre-balancing for 15 min, spreading upward at a spreading distance of 8cm, taking out, air drying, placing under an ultraviolet lamp (365nm) for inspection, and displaying the main fluorescence spots with the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the reference solution. From the chromatogram analysis of the lamella plate, the fat-soluble information component is observed. It has also been reported【2】Chromatograms of 2 batches of eclipta medicinal materials are shown, but no method and developing agent are reported, and the inspection condition is that 4 fluorescent spots are shown on the thin-layer plate under the ultraviolet lamp of 365 nm. From the spot analysis of the chromatogram, it belongs to fat-soluble components. The gradient full-information thin-layer identification method and the identification chromatogram of the yerbadetajo herb are not referred yet.
The method provides a simple, convenient, quick, low-cost and high-efficiency multi-information identification method for the eclipta medicinal material and various preparations prepared by different processes, and performs special multi-information gradient thin-layer identification research on the eclipta medicinal material. A set of simple, convenient and quick holographic eclipta thin-layer identification method and a spectrum without environmental pollution are obtained.
Disclosure of Invention
A test sample solution and a reference medicinal material solution are obtained by a simple and quick pretreatment method, and about twenty-seven information spots of the eclipta medicinal material are inspected on 4 thin-layer plates by adopting the same test sample solution and 4 developing agents with different polarities under various inspection conditions. The components are crossed with each other, but do not interfere with each other under the respective inspection conditions, and clear fluorescent spots or color spots can be displayed. The thin-layer gradient identification is completed by 0.2g of each of the medicinal material and the reference medicinal material, 4ml of extraction solvent, 40ml of developing solvent and 1.5 hours. Simple, convenient, quick and efficient, and is not available in the reported methods at present. Provides a method and a chromatogram map for multi-information quick quality supervision of the yerbadetajo herb medicine. Provides thin-layer identification reference information for the eclipta compound preparation prepared by different preparation processes.
The technical scheme adopted by the invention for solving the technical problems is as follows:
(1) carrying out thin-layer identification on fat-soluble components of the eclipta medicinal material, grinding 0.2g of the eclipta medicinal material, adding 2ml of methanol, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernate as a test solution; preparing 0.2g of herba Ecliptae control medicinal material, and preparing control medicinal solution by the same method; respectively dripping 4-5 μ l of each of the test solution and the reference solution on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate as developing agent at volume ratio of 19: 1, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution;
(2) thin-layer identification of fat-soluble components of herba Ecliptae medicinal material by absorbing 4-5 μ l of each of the sample solution and the reference medicinal material solution under item (1), and respectively dropping on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 12: 3: 0.5, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, inspecting under ultraviolet lamp 365nm to obtain the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the control solutionA primary spot of colored fluorescence; inspecting in sunlight, wherein spots of the same color appear in the chromatogram of the test solution at the positions corresponding to those in the chromatogram of the reference solution;
(3) thin-layer identification of polar components in eclipta medicinal material, absorbing 4-5 mul of each of the test solution and the reference medicinal material solution under the item (1), and respectively dropping the test solution and the reference medicinal material solution on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 6: 4: 0.5, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution;
(4) thin-layer identification of water-soluble components of eclipta medicinal materials, namely, respectively dropping 4-5 mul of test solution and 5 mul of reference medicinal material solution under the item (1) on the same silica gel GF254Developing on a thin layer plate with chloroform-methanol-formic acid-water as developing agent at volume ratio of 10: 3: 0.2: 0.3, taking out, air drying, and inspecting under 365nm ultraviolet lamp; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; 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 fluorescent spots with the same color in the sample chromatogram and the corresponding position of the control medicinal material chromatogram; then the sample is placed in a dark room and inspected by lamplight, and spots with the same color appear in the chromatogram of the sample at the positions corresponding to the chromatograms of the reference medicinal materials.
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, the developing agents with different polarities are adopted for development, and various chemical components can be well separated on respective thin-layer plates along with different developing agents according to different adsorption, desorption, re-adsorption and re-desorption capacities. 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 do not interfere with each other on different layers, so that different spot colors are presented, and a thin-layer chromatogram with multiple information is obtained. And according to the polarity of the developing agent, well-separated multi-information gradient thin-layer chromatograms on each thin-layer plate are connected from non-polarity to form a set of holographic thin-layer chromatograms.
The invention has the following innovation points and beneficial effects:
1. developing liposoluble components with cyclohexane-ethyl acetate at volume ratio of 19: 1, and inspecting thin layer plate under ultraviolet lamp 365nm to obtain 6 clear fluorescent spots, 4 of which are bluish purple, 1 of which is bright yellow green, and 1 of which is light yellow green (FIG. 1); placing the film under an ultraviolet lamp at 254nm to obtain spot-free information; no clear spots were observed after development by spraying with 10% sulfuric acid. The developing agent can obtain more fat-soluble spots and better definition.
2. Taking cyclohexane-ethyl acetate-formic acid with a volume ratio of 12: 3: 0.5 as a developing agent, and inspecting 12 fluorescent spots with different colors by using developed fat-biased soluble components under an ultraviolet lamp at 365nm, wherein 6 fluorescent spots have the same color with the fat-soluble components, the Rf value is more than 0.5, and 2 red fluorescent spots, 1 blue-violet fluorescent spot and 3 pink fluorescent spots are presented below 0.5 (figure 2); spraying 10% ethanol sulfate solution for color development, and placing under 365nm ultraviolet lamp for inspection to show 6 clear fluorescent spots, 2 of which are light pink, 2 light green, 1 orange yellow and 1 orange red (figure 3); obviously, the amount and the color of the fluorescent spots are different from those of spots before color development, the fluorescent spots are non-fluorescent compounds, the fluorescent spots are excited after the fluorescent spots react with 10% sulfuric acid, and the fluorescent spots before color development and the fluorescent spots after color development are not the same compounds; the developed thin layer plates were further examined in the sunlight to show 5 dark and light brown spots (FIG. 4), wherein 4 spots were present, and the spots were different from the fluorescent spots in color in Rf value and spot size, and were not fluorescent before and after development. As calculated above, the same lamella plate detects about 21 different information spots under three viewing conditions, and the new detection spot is 15 by subtracting 6 spots repeated with fat solubility. Under different inspection conditions, different spots are inspected, although the spots are overlapped with each other, under the respective inspection conditions, the spots are not interfered with each other, and information complementation and reinforcement are achieved. In the thin layer identification of fat-soluble components, the information obtained by the identification is the largest, and the separation degree between spots is also better, which is reported for the first time.
3. Developing the medium-polarity components with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 6: 4: 0.5 to obtain 9 fluorescent spots of different colors detected under ultraviolet lamp 365nm, wherein 3 of the fluorescent spots are bluish purple, 2 bright blue, 1 bright yellow green, 2 red and 1 light pink (FIG. 5); compared with the thin-layer chromatogram of the fat-soluble component, 2 bright blue fluorescent spots are new identification characteristic spots.
4. Developing water soluble components with chloroform-methanol-formic acid-water as developing agent at volume ratio of 10: 3: 0.2: 0.3 to obtain 3 new bright blue fluorescent spots (figure 6) detected under 365nm ultraviolet lamp; comparing the thin layer chromatogram analysis of the polar components, wherein 2 fluorescence spots with larger Rf values are reported under item (3), and 1 fluorescence spot with small Rf value is a new spot of the water-soluble components; spraying 10% ethanol sulfate solution to develop color, placing under 365nm ultraviolet lamp to inspect, presenting 2 clear fluorescent spots, analyzing from Rf value, all different from directly inspected fluorescent spots, not the same compound (figure 7); the thin layer plate was placed in a dark room again and inspected by light to show 1 main tan spot which had a different Rf value from the fluorescent spots before and after development and was not the same compound. Thus, 6 water-soluble components were detected in total, and 4 of them were new component spots.
5. About twenty-seven information spots of the eclipta medicinal material are inspected on 4 thin-layer plates by adopting the same test solution and the reference medicinal material solution and 4 developing agents with different polarities under various inspection conditions. Some spots which are left on the upper surface can be clearly identified on each thin-layer plate, and the spots are mutually crossed but do not interfere with each other under respective inspection conditions, so that clear fluorescent spots or color spots can be presented. Provides a method and a map for the quality evaluation and quality comparison of the yerbadetajo herb medicines with different sources, different production places and different harvesting seasons; provides an optional multi-information chromatogram map for compound preparations with different processes and compositions when carrying out thin-layer identification on the yerbadetajo herb.
6. The present identification method is also different from the reported methods in that: the non-polar, medium-polar and polar components of the developing solvent are all acidic, and the reported methods are all neutral. The acidic developing agent enables some acidic substances such as flavonoid and thiophene components to exist in a molecular form, overcomes the trailing phenomenon of polyphenol hydroxyl compounds, enables all spots to shrink and concentrate, enables more than ten fluorescent spots with different colors to present respective clear color boundaries, and is beneficial to observation and judgment.
Drawings
FIG. 1 is a thin-layer TLC (thin-layer chromatography) chart of fat-soluble ingredients of herba Ecliptae medicinal materials under an ultraviolet lamp of 365 nm.
FIG. 2 is a thin-layer TLC (thin-layer chromatography) chart of fat-soluble components of herba Ecliptae medicinal material under 365nm ultraviolet lamp.
FIG. 3 is a TLC image of the fat-soluble components of Ecliptae herba under 365nm ultraviolet light after developing with 10% ethanol sulfate solution.
FIG. 4 is a TLC image of the thin layer observed in the sun after the relatively fat-soluble components of the yerbadetajo herb are developed by 10% sulfuric acid ethanol solution.
FIG. 5 is a thin-layer TLC chart of the polar components in the eclipta medicinal material under the ultraviolet lamp 365 nm.
FIG. 6 is a thin-layer TLC chart of water-soluble components of Ecliptae herba under 365nm ultraviolet lamp.
FIG. 7 is a TLC image of the water-soluble components of Ecliptae herba under 365nm ultraviolet light after being developed with 10% ethanol sulfate solution.
FIG. 8 is a TLC chart of the thin layer light inspection of water-soluble components of eclipta medicinal materials after being developed with 10% sulfuric acid ethanol solution.
In fig. 1, 1 eclipta control drug; 2.3.4.5 is herba Ecliptae.
FIGS. 2, 3 and 4 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1 is herba Ecliptae reference material; 2.3.4.5 is herba Ecliptae.
In fig. 5, 1 eclipta control drug; 2.3.4.5 is herba Ecliptae.
FIGS. 6, 7 and 8 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1 is herba Ecliptae reference medicinal material; 2.3.4.5 is herba Ecliptae.
The specific implementation mode of the invention is as follows:
(1) carrying out thin-layer identification on fat-soluble components of the eclipta medicinal material, grinding 0.2g of the eclipta medicinal material, adding 2ml of methanol, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernate as a test solution; preparing 0.2g of herba Ecliptae control medicinal material, and preparing control medicinal solution by the same method; respectively dripping 4-5 μ l of each of the test solution and the reference solution on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate as developing agent at volume ratio of 19: 1, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution;
(2) thin-layer identification of fat-soluble components of herba Ecliptae medicinal material by absorbing 4-5 μ l of each of the sample solution and the reference medicinal material solution under item (1), and respectively dropping on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 12: 3: 0.5, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clear, and inspecting under ultraviolet lamp 365nm to obtain main fluorescent spots with the same color in the sample chromatogram and the corresponding position of the control chromatogram; inspecting in sunlight, wherein spots of the same color appear in the chromatogram of the test solution at the positions corresponding to those in the chromatogram of the reference solution;
(3) thin-layer identification of polar components in eclipta medicinal material, absorbing 4-5 mul of each of the test solution and the reference medicinal material solution under the item (1), and respectively dropping the test solution and the reference medicinal material solution on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 6: 4: 0.5, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; for supplying toIn the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution;
(4) thin-layer identification of polar components in eclipta medicinal material, absorbing 4-5 mul of each of the test solution and the reference medicinal material solution under the item (1), and respectively dropping on the same silica gel GF254Developing on a thin layer plate with chloroform-methanol-formic acid-water as developing agent at volume ratio of 10: 3: 0.2: 0.3, taking out, air drying, and inspecting under 365nm ultraviolet lamp; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; 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 fluorescent spots with the same color in the sample chromatogram and the corresponding position of the control medicinal material chromatogram; then the sample is placed in a dark room and inspected by lamplight, and spots with the same color appear in the chromatogram of the sample at the positions corresponding to the chromatograms of the reference medicinal materials.
Reference to the literature
【1】 Chromatogram gallery-Chinese medicinal material-thin layer chromatography-eclipta-different thin layer plate thin layer chromatogram comparison (1) - (6)
【2】 The document "2 batches of eclipta thin layer chromatograms" uploaded to hundred degree libraries by grandchild flower 2016.1.14.
Claims (2)
1. A gradient full-information thin-layer identification method for an eclipta medicinal material is characterized by comprising the following steps of:
(1) carrying out thin-layer identification on fat-soluble components of the eclipta medicinal material, grinding 0.2g of the eclipta medicinal material, adding 2ml of methanol, carrying out ultrasonic treatment for 15 minutes, centrifuging, and taking supernate as a test solution; preparing 0.2g of herba Ecliptae control medicinal material, and preparing control medicinal solution by the same method; respectively dripping 4-5 μ l of each of the test solution and the reference solution on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate as developing agent at volume ratio of 19: 1, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution;
(2) thin-layer identification of fat-soluble components of herba Ecliptae medicinal material under item (1)4-5 mul of each of the test solution and the reference solution are respectively spotted on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 12: 3: 0.5, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots are clear, and inspecting under ultraviolet lamp 365nm to obtain main fluorescent spots with the same color in the sample chromatogram and the corresponding position of the control chromatogram; inspecting in sunlight, wherein spots of the same color appear in the chromatogram of the test solution at the positions corresponding to those in the chromatogram of the reference solution;
(3) thin-layer identification of polar components in eclipta medicinal material, absorbing 4-5 mul of each of the test solution and the reference medicinal material solution under the item (1), and respectively dropping the test solution and the reference medicinal material solution on the same silica gel GF254Developing on the thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 6: 4: 0.5, taking out, air drying, and inspecting under ultraviolet lamp 365 nm; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution;
(4) thin-layer identification of polar components in eclipta medicinal material, absorbing 4-5 mul of each of the test solution and the reference medicinal material solution under the item (1), and respectively dropping on the same silica gel GF254Developing on a thin layer plate with chloroform-methanol-formic acid-water as developing agent at volume ratio of 10: 3: 0.2: 0.3, taking out, air drying, and inspecting under 365nm ultraviolet lamp; in the chromatogram of the test solution, the main fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution; 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 fluorescent spots with the same color in the sample chromatogram and the corresponding position of the control medicinal material chromatogram; then the sample is placed in a dark room and inspected by lamplight, and spots with the same color appear in the chromatogram of the sample at the positions corresponding to the chromatograms of the reference medicinal materials.
2. The gradient complete information thin-layer identification method of the eclipta medicinal material as claimed in claim 1, characterized in that about 27 characteristic spots are detected on 4 thin-layer plates with 4 different developing agents, the time is about 1.5 hours, the extraction solvent is 4ml, and the developing agent is 40 ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911069548.0A CN110824094B (en) | 2019-10-30 | 2019-10-30 | Gradient full-information thin-layer identification method for eclipta medicinal materials |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911069548.0A CN110824094B (en) | 2019-10-30 | 2019-10-30 | Gradient full-information thin-layer identification method for eclipta medicinal materials |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110824094A true CN110824094A (en) | 2020-02-21 |
| CN110824094B CN110824094B (en) | 2021-06-01 |
Family
ID=69552355
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911069548.0A Active CN110824094B (en) | 2019-10-30 | 2019-10-30 | Gradient full-information thin-layer identification method for eclipta medicinal materials |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110824094B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112051353A (en) * | 2020-09-23 | 2020-12-08 | 浙江金大康动物保健品有限公司 | Gradient full-information thin-layer identification method for radix peucedani medicinal material |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060110479A1 (en) * | 2004-11-24 | 2006-05-25 | Mitra Shankar K | Natural composition for curing hepatitis-B, methods for making the same and pharmaceutical formulations thereof |
| CN101028487A (en) * | 2007-04-04 | 2007-09-05 | 西安碑林药业股份有限公司 | Chinese-medicinal preparation for treating eyeground bleeding and method for inspecting vision-improving prescription quality |
| CN101363819A (en) * | 2008-01-08 | 2009-02-11 | 广西博科药业有限公司 | Quality control method for yuye qinghuo capsule |
| CN102210781A (en) * | 2011-06-01 | 2011-10-12 | 中山大学附属第一医院 | Preparation method and quality control method of traditional Chinese medicine Danmo capsule |
-
2019
- 2019-10-30 CN CN201911069548.0A patent/CN110824094B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060110479A1 (en) * | 2004-11-24 | 2006-05-25 | Mitra Shankar K | Natural composition for curing hepatitis-B, methods for making the same and pharmaceutical formulations thereof |
| CN101028487A (en) * | 2007-04-04 | 2007-09-05 | 西安碑林药业股份有限公司 | Chinese-medicinal preparation for treating eyeground bleeding and method for inspecting vision-improving prescription quality |
| CN101363819A (en) * | 2008-01-08 | 2009-02-11 | 广西博科药业有限公司 | Quality control method for yuye qinghuo capsule |
| CN102210781A (en) * | 2011-06-01 | 2011-10-12 | 中山大学附属第一医院 | Preparation method and quality control method of traditional Chinese medicine Danmo capsule |
Non-Patent Citations (5)
| Title |
|---|
| MITESH D. PHALE ET AL: "Precise and sensitive HTPLC method for quantitative estimation of wedelolactone in Eclipta Alba hassk", 《PHARMACOPHORE》 * |
| SOMESHI THAPLIYAL ET AL: "Analysis of wedelolactone in Eclipta Alba and its fomulation by HPTLC", 《JOURNAL OF GLOBAL TRENDS IN PHARMACEUTICAL SCIENCES》 * |
| 张朝凤 等: "《2010全国知名中医院院长暨道家文化与中医药养生论坛论文集》", 31 December 2010 * |
| 林启焰 等: "滋肾养阴颗粒的质量标准研究", 《中草药》 * |
| 梁丽娟 等: "中药墨旱莲的质量标准研究", 《国际中医中药杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112051353A (en) * | 2020-09-23 | 2020-12-08 | 浙江金大康动物保健品有限公司 | Gradient full-information thin-layer identification method for radix peucedani medicinal material |
| CN112051353B (en) * | 2020-09-23 | 2022-10-11 | 浙江金大康动物保健品有限公司 | Gradient full-information thin-layer identification method for radix peucedani medicinal material |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110824094B (en) | 2021-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111487361B (en) | Gradient full-information thin-layer identification method for rhizoma atractylodis medicinal material | |
| CN111024834B (en) | Gradient full-information thin-layer identification method for notopterygium root medicinal material | |
| Pascual et al. | Simplified screening by TLC of plant drugs | |
| CN108956845B (en) | One-plate four-medicine multi-information thin-layer identification method for three-ingredient decoction freeze-dried powder | |
| CN110579560B (en) | Gradient multi-information thin-layer identification method for poplar flower medicinal materials | |
| CN110221015B (en) | Gradient full-information thin-layer identification method for pomegranate bark medicinal material | |
| CN110376326B (en) | Multi-region thin layer identification and distinguishing method for pericarpium citri reticulatae and immature bitter orange medicinal materials | |
| CN110824094B (en) | Gradient full-information thin-layer identification method for eclipta medicinal materials | |
| CN112051353B (en) | Gradient full-information thin-layer identification method for radix peucedani medicinal material | |
| CN111060647B (en) | Gradient full-information thin-layer identification method for medicated leaven medicinal materials | |
| CN107727787B (en) | A kind of TLC Identification identifying hymsleya amabilis kind | |
| CN110426487A (en) | A kind of quick thin-layer identification method of multi information of thick wood-fern rhizome medicinal material | |
| CN108956843B (en) | Rapid multi-information thin-layer identification method for pinellia ternate, bighead atractylodes rhizome and gastrodia elata decoction freeze-dried powder | |
| CN111024876B (en) | Gradient full-information thin-layer identification method for glossy privet fruit medicinal materials | |
| CN101829176A (en) | Method for identifying rhizoma atractylodis macrocephalae by adopting thin layer chromatography | |
| CN105277648A (en) | Multi-information gradient thin layer discriminating method of Radix Puerariae medicinal material and water extract product thereof | |
| CN102608250B (en) | Rapid detection method for Jinfangganmao granules | |
| CN106370766A (en) | Method for identifying atractylodes macrocephala koidz in donkey-hide glue blood-enriching preparation | |
| CN112305142A (en) | Gradient multi-information thin-layer identification method for bighead atractylodes rhizome medicinal material | |
| CN104792920B (en) | The inverse thin layer chromatography discrimination method of bile acids composition in a kind of poultry class gallbladder powder | |
| CN114324725B (en) | Gradient full-information thin-layer identification method for Zhaoshan white extract tablet | |
| CN108663472B (en) | Identification method of rhododendron anthopogonoides essential oil | |
| CN112268965B (en) | Multi-region thin layer identification and distinguishing method for frankincense and myrrh medicinal materials | |
| CN104991032A (en) | Thin layer chromatography for identifying doped kadsura longepedunculata in Chinese magnoliavine | |
| CN108693295B (en) | Identification method of artemisia pigweed essential oil |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A gradient full-information TLC identification method for Eclipta chinensis Effective date of registration: 20230223 Granted publication date: 20210601 Pledgee: Bank of Jinhua Limited by Share Ltd. science and Technology Branch Pledgor: ZHEJIANG DKCOM ANIMAL HEALTH PRODUCTS Co.,Ltd. Registration number: Y2023980033287 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20231108 Granted publication date: 20210601 Pledgee: Bank of Jinhua Limited by Share Ltd. science and Technology Branch Pledgor: ZHEJIANG DKCOM ANIMAL HEALTH PRODUCTS Co.,Ltd. Registration number: Y2023980033287 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A gradient full information thin-layer identification method for the medicinal herb of Echinochloa grandiflorus Effective date of registration: 20231114 Granted publication date: 20210601 Pledgee: Bank of Jinhua Limited by Share Ltd. science and Technology Branch Pledgor: ZHEJIANG DKCOM ANIMAL HEALTH PRODUCTS Co.,Ltd. Registration number: Y2023980065314 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |