AU2020100587A4 - Method For Extracting Oligomeric Polyphenol Compounds From Plant Seeds - Google Patents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
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- A—HUMAN NECESSITIES
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- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
- A61K31/343—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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Abstract
Abstract The invention discloses a method for extracting oligomeric polyphenolic compounds from plant seeds, the method comprising the steps of: crushing the plant seeds, soaking the plant seeds with ethanol to obtain a leachate, distilling under reduced pressure to obtain an ethanol extractum, dissolving the ethanol extractum, decoloring, eluting to obtain an eluant, distilling the eluant under reduced pressure to obtain an extractum, and separating by high performance liquid chromatography to obtain the oligomeric polyphenolic compound. According to the method, while the polyphenol function is kept, the extraction rate is increased, and the oligomeric polyphenol compounds can be used for preparing gram-positive drug-resistance bacterium drugs, additives, health products, food and other products.
Description
Technical Field
The invention relates to the technical field of extraction of plant active components, in particular to a method for extracting oligomeric polyphenolic compounds from plant seeds.
Background Art
Polyphenol is a plant polyphenol active substance, which is insoluble in water and soluble in organic solvents such as methanol, ethanol and acetone. It is widely found in plant seeds, seed coats and fruit juice. Such polyphenol consists of phenolic acids such as epicatechin, flavanols, anthocyanins, flavonols, condensed tannins, resveratrol oligomers and the like, wherein the 15 content of pro anthocyanidins is the most abundant and can reach 80-85%, followed by other components such as catechin and epicatechin reaching about 5%. Various components in the polyphenol have different contents, such that their colors vary from deep rose to light brownish red. At present, most domestic and foreign polyphenol extraction methods for plant seeds are decoction methods with the characteristics of simplicity in operation, low cost and the like, but 20 this method easily destroy polyphenols and other antioxidant components in the plant seeds, and an extracting solution has a high content of impurities, which is to the disadvantage of separation and purification of polyphenols. Therefore, it is necessary to find a method for preventing automatic oxidation of polyphenols in plant seeds in an extraction process so as to ensure the stability of the activity of polyphenols.
Summary of the Invention
The invention aims to provide a method for extracting oligomeric polyphenolic compounds from plant seeds so as to solve the problems of the prior art.
In order to achieve the above purpose, the invention provides the following scheme.
The invention provides a method for extracting oligomeric polyphenolic compounds from plant seeds, comprising the following steps of:
(1) crushing the plant seeds, soaking the plant seeds in ethanol with the volume concentration of
70% at normal temperature for 3 days, and repeating twice to obtain a leachate;
(2) distilling the obtained lixivium under reduced pressure to obtain an ethanol extractum;
(3) after dissolving the ethanol extractum with a solvent, taking and eluting macroporous resin by a solvent method for decolorization; after concentrating a collected mixed eluant, mixing with silicon (100-200 meshes) for samples, packing, eluting with different eluents respectively, and distilling the eluents under reduced pressure respectively to obtain ethyl acetate extractum and acetone extractum respectively; and (4) taking an acetone extract, separating by high performance liquid chromatography to obtain the oligomeric polyphenol compound, and further to obtain dimeric and trimeric compounds of oligomeric polyphenols when adopting gradient elution with methanol and water according to ratios.
Further, the feed-liquid weight ratio (W/W) of the plant seeds to ethanol in the step (1) is 1:3-4.
Further, the solvent in the step (3) is water, and the eluant are gradient eluants of water and ethanol according to ratios (9:1-7:3-5:4-1:1-3:7-1:9-0:10), and the mixed eluant is collected from fractions of the gradient elution with water and ethanol according to ratios (3:7-1:9-0:10).
Further, the eluting process in the step (3) is performed by using 5-8 times of column volume of ethyl acetate, and then performed by using 5-8 times of column volume of acetone, to obtain ethyl acetate extractum and acetone extractum, respectively.
Further, high performance liquid chromatography in the step (4) is performed, under the conditions of a ratio of methanol to water being 55:45, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm, to obtain components Fl, F2, F3 and F4, the component Fl is subjected to the high performance liquid chromatography, under the separation conditions for the component Fl of a ratio of methanol to water being 50:50, a flow rate of 8 mL/min, YMC ODS C18, 250*20 mm, to obtain a compound gentian H; and the component F3 is subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 45:55, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain a compound trans-D-viniferin.
The invention also provides oligomeric polyphenolic compounds prepared according to the preparation method, wherein the compounds are trans-D-viniferin and trans-gentian H. The compounds can be used for preparing gram-positive drug-resistance bacterium products.
The invention has the following technical effects.
By adopting an effective part extraction and separation method, oligomeric polyphenol can be directly produced from seeds of various plants, the extraction rate of polyphenol is increased while the polyphenol function is kept, and the extraction rate of the oligomeric polyphenol compounds extracted by the method is high, reaching 1.5-1.8%.
According to the method defined by the invention, a large amount of impurities such as wax fat, starch and protein are effectively removed and separated in the production process, so that the purity of the polyphenol compound produced by extraction is high.
The drug-resistant bacterium spectrums of the two compounds, namely trans-D-viniferin and trans-gentian H, prepared by the method are gram-positive drug-resistant bacteria, and the extracted oligomeric dimeric and trimeric phenolic compounds will be widely applied to preparation of gram-positive drug-resistance bacterium drugs, additives, health products, food and other products.
Detailed Description of the Invention
Various exemplary embodiments of the present invention will now be described in detail, which should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features and embodiments of the present invention.
It is understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. Further, for the numerical range in the present invention, it is understood that the present invention also specifically discloses every value between the upper and lower limits of the range. Each smaller range between any stated value or stated range and any other stated value or stated range is also included in the present invention. The upper and lower limits of these smaller ranges may independently be included or excluded from the range.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present invention without departing from the scope or spirit of the present invention. Other embodiments derived from the description of the present invention will be apparent to those skilled in the art. The present description and embodiments are exemplary only.
Embodiment 1
Take and soak 7 kg grape seeds in ethanol with the volume concentration of 70% at normal temperature for 3 days, repeat twice to obtain a leachate, and distill the obtained leachate under reduced pressure to obtain 400 g ethanol extractum. Take 300 g ethanol extractum, dissolve the ethanol extractum with a solvent, take macroporous resin for decolorization, then mix with 800 g silica gel (100-200 meshes) for samples and packing. Elute by using ethyl acetate with the volume of 5 times of column volume, then elute by using acetone with the volume of 5 times of column volume, and distill eluants under reduced pressure, respectively, to obtain 80 g ethyl acetate extractum and 87 g acetone extractum.
Take an acetone extract, and perform high performance liquid chromatography of Waters (a ratio of methanol to water being 55: 45, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain components Fl, F2, F3 and F4. The component Fl was subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 50: 50, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain a compound trans-gentian H. The extraction rate of trans-gentian H in this embodiment was 1.80%. Component F3 was subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 45: 55, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain a compound trans-D-viniferin. The extraction rate of trans-D-viniferin in this embodiment was 1.70%.
Embodiment 2
Take and dry 7 kg paeonia suffruticosa seeds and remove husks. Soak with 70% ethanol at room temperature for 3 days, repeat twice, and distill an obtained leachate under reduced pressure to obtain 400 g ethanol extractum. Take 300 g ethanol extractum, dissolve the ethanol extractum with a solvent, take macroporous resin for decolorization, then mix with 800 g silica gel (100-200 meshes) for samples and packing. Elute first with 8 times of column volume of ethyl acetate and then with 8 times of column volume of acetone. Distill eluants under reduced pressure, respectively, to obtain 110 g ethyl acetate extractum and 150 g acetone extractum.
Take an acetone extract, perform high performance liquid chromatography of Waters (a ratio of methanol to water being 55: 45, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain components Fl, F2, F3 and F4. The component Fl was subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 50: 50, a flow rate of 8 mL/min, YMC ODS C18, 250*20 mm) to obtain trans-gentian H; the component F3 was subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 45: 55, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain trans-D-viniferin.
Embodiment 3
Take and dry 7 kg paeonia delavayi seeds and remove husks. Soak with 70% ethanol at room temperature for 3 days, repeat twice, and distill an obtained leachate under reduced pressure to obtain 400 g ethanol extractum. Take 300 g ethanol extractum, dissolve the ethanol extractum with a solvent, take macroporous resin for decolorization, then mix with 800 g silica gel (100-200 meshes) for samples and packing. Elute first with 7 times of column volume of ethyl acetate and then with 7 times of column volume of acetone. Distill eluants under reduced pressure, respectively, to obtain 110 g ethyl acetate extractum and 150 g acetone extractum.
Take an acetone extract, perform high performance liquid chromatography of Waters (a ratio of methanol to water being 55: 45, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain components Fl, F2, F3 and F4. The component Fl was subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 50: 50, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain trans-gentian H; the component F3 was subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 45: 55, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain trans-D-viniferin.
Assay for drug-resistance bacteria for compound trans-gentian H and trans-D-viniferin:
1. Preparation of antibacterial drug stock solution
The positive control antibiotic vancomycin and all strains were provided by the Institute of Medical Biotechnology, Beijing Chinese Academy of Medical Sciences.
With reference to operation standards for antimicrobial susceptibility test from NCCLS, the preparation is performed specifically, and the concentrations of two oligomeric phenols, trans-D-viniferin and trans-gentian H, should not be less than 1000 Lig/mL. The amount of antibiotic powder was accurately weighed using an analytical balance to be 171.2 mg. The amount of diluent required was calculated according to the formula: (171.2mgx750pg/mL)/1200pg/mL=107.0mL, then 171.2 mg antibiotic powder was dissolved in 107.0 mL diluent. The prepared antibacterial drug stock solution should be stored under -60 °C environment, and the storage period does not exceed 4-6 months.
2. Preparation of culture medium for drug sensitivity test
The culture medium used Mueller-Hinton (MH) broth, pH 7.20-7.40, according to the operation standards for Antimicrobial Susceptibility Test from NCCLS. Additional 2% (W/V) sodium chloride was added to the broth. The required amount of MH broth was prepared as required by a manufacturer.
3. Preparation of inoculum by direct colony suspension formulation. Methicillin-resistant staphylococcus aureus were directly cultured for 18-24 h. Colonies of methicillin-resistant staphylococcus aureus were taken by 1-2 and placed in the broth culture solution with the pH value of 7.20-7.40 in the above Item 2, and prepared into a bacterial suspension of 0.5 McFarland standard.
The bacterial suspension was distilled with MH broth, according to a ratio of the bacterial suspension to the MH broth being 1:1, for use. It is noted that the prepared inoculum should be inoculated within 15 minutes and one portion of inoculum was subcultured on a non-selective agar plate to check the purity of the inoculum.
4. Preparation of diluted antibacterial drug and inoculation of bacterial solution sterile test tubes (13x100 mm) were arranged in a row; except that 1.60 mL MH broth was added to the first tube, 1.00 mL MH broth was added to each of the other tubes; 0.40 mL trans-d-viniferin antibacterial drug stock solution (such as 1.00 pg/mL) was added to the first tube and mixed uniformly; then 1.00 mL solution was sucked from the first tube to the second tube; after mixing uniformly, 1.00 mL solution was sucked from the second tube to the third tube; dilution was continuously performed in such a way till the 11th tube; and 1.00 mL solution was sucked from the 11th tube and discarded; and the 12th tube was a growth control without drug. At this point, the drug concentrations of the tubes were 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25
2020100587 17 Apr 2020 pg/mL, respectively. 1.00 mL prepared inoculum was then added to each tube to make a final concentration of bacterium solution about 5xlO5CFU/mL per tube. Tubes 1 through 11 had drug concentrations of 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125 pg/rnL, respectively. Similarly, similar operations were carried out on the trans-gentian H oligomeric phenolic compound and positive 5 control vancomycin as above. The inoculated dilution tubes were stoppered and incubated for 24 h in a 35 °C common air incubator.
5. Result judgment and explanation
Before the MIC of the tested strain was read and reported, whether the bacterial growth situation of the growth control tube was good should be checked, meanwhile, whether the subculture situation of the inoculum should also be checked to determine whether it was polluted, and whether the MIC value of the quality-control strain was in a quality-control range. The MIC of the tested bacteria is determined by naked eye observation when there is no bacterial growth in the tube with the lowest drug concentration. Compared with the positive growth control tube, the drug concentration of the growth tube which inhibited 80% of bacteria is the MIC value of the tested bacteria.
The MIC value test results of the two compounds obtained by the invention against drug-resistant bacteria are shown in table 1.
Table 1
| Drug number | Trans-D-viniferin | Trans-gentian H | Vancomycin | |||
| Strain name | Strain number | Drug resistance | Controls | |||
| Staphylococcus epidermidis | Staphylococcus epidermidis | ATCC 12228 | MSSE | 8 | 16 | 0.5 |
| Staphylococcus epidermidis | 43843 | MSSE | 8 | 16 | 1 | |
| Staphylococcus epidermidis | 43903 | MRSE | 32 | 16 | 0.5 | |
| Staphylococcus aureus | Staphylococcus aureus | ATCC 29213 | MSSA | 8 | 32 | 0.5 |
| Staphylococcus aureus | ATCC 33591 | MRSA | 8 | 32 | 0.5 | |
| Staphylococcus aureus | 15 | MSSA | 16 | 32 | 0.5 | |
| Staphylococcus aureus | 13 -17 | MSSA | 16 | 32 | 0.5 | |
| Staphylococcus aureus | 13 -18 | MRSA | 16 | 32 | 0.5 | |
| Enterococcus faecalis | Enterococcus faecalis | ATCC 29212 | VSE | 32 | 32 | 2 |
| Enterococcus faecalis | ATCC | VRE | 64 | 32 | 8 |
2020100587 17 Apr 2020
51299
ATCC
| Enterococcus faecalis Enterococcus faecalis | 51575 43934 | VRE VSE | 64 32 | 128 32 | 128 1 | |
| Enterococcus faecium | Enterococcus faecium | ATCC 700221 | VRE | 128 | 32 | >128 |
| Enterococcus faecium | 44025 | VSE | 128 | 32 | 0.5 | |
| Enterococcus faecium | 44166 | VRE | 32 | 16 | 32 | |
| Escherichia coli | Escherichia coli | ATCC 25922 | ESBLs(-) | >128 | >256 | >128 |
| Escherichia coli | 1515 | ESBLs(-) | >128 | >256 | >128 | |
| Escherichia coli | 44118 | ESBLs(-) | >128 | 256 | >128 | |
| Escherichia coli | 44149 | ESBLs(+) | >128 | >256 | >128 | |
| Klebsiella pneumoniae | Klebsiella pneumoniae | ATCC 700603 | ESBLs(+) | >128 | >256 | >128 |
| Klebsiella pneumoniae | 7 | ESBLs(-) | >128 | >256 | >128 | |
| Klebsiella pneumoniae | ATCC BAA-2146 | NDM-1(+) | >128 | 256 | >128 | |
| Klebsiella pneumoniae | 43935 | ESBLs(-) | >128 | >256 | >128 | |
| Klebsiella pneumoniae | 14 -15 | ESBLs(+) | >128 | >256 | >128 | |
| Pseudomonas aeruginosa | Pseudomonas aeruginosa | ATCC 27853 | >128 | >256 | >128 | |
| Pseudomonas aeruginosa | PAO1 | 128 | 256 | >128 | ||
| Pseudomonas aeruginosa | 13 -46 | >128 | >256 | >128 | ||
| Acinetobacter calcoacetious | Acinetobacter calcoacetious | ATCC 19606 | >128 | >256 | >128 | |
| Enterobacter cloacae | Enterobacter cloacae | ATCC 43560 | 128 | 256 | >128 | |
| Enterobacter aerogenes | Enterobacter aerogenes | ATCC 13048 | >128 | >256 | >128 | |
| Serratiamarcescens | Serratiamarcescens | ATCC 21074 | >128 | >256 | >128 | |
| Morganellamorganii | Morganellamorganii | ATCC 25830 | >128 | >256 | >128 | |
| Providentiarettgeri | Providentiarettgeri | ATCC 31052 | >128 | >256 | >128 | |
| Proteus vulgaris | Proteus vulgaris | ATCC 29905 | >128 | >256 | >128 | |
| Proteus mirabilis | Proteus mirabilis | 43843 | >128 | >256 | >128 | |
| Stenotrophomonasmaltophilia | Stenotrophomonasmaltophilia | ATCC 13636 | 128 | >256 | >128 | |
| Citrobacterfreundii | Citrobacterfreundii | ATCC 43864 | >128 | >256 | >128 |
2020100587 17 Apr 2020
As can be seen from Table 1, 1) the concentrations of the compound trans-D-viniferin methicillin-resistant staphylococcus aureus (MRSA) and methicillin-sensitive staphycoccus epidermidis (MSSE) were both 8 pg/mL; the concentration of the trans-gentian H was 16 pg/mL, and the concentration of the methicillin-resistant staphylococcus aureus (MRSA) was 32 pg/mL.
They all showed strong activity against gram-positive drug-resistance bacteria, and the concentrations were greater than 128 pg/mL for all gram-negative drug-resistance bacteria, so that they have no activity against the gram-negative drug-resistance bacteria. 2) From trans-D-viniferin to trans-gentian H, it is actually the change from a dimer to a trimer with the water solubility decreased and the activity of drug-resistance bacteria weakened. Therefore, the trans-D-viniferin 10 dimer has stronger resistance to drug-resistance bacteria. 3) Through comparative research on the structure-activity relationship of the two compounds showed that both trans-D-viniferin and trans-gentian H were trans-configurations; the opposite cis-configurations, whether the trans-D-viniferin or the trans-gentian H, showed no activity against drug-resistance bacteria.
The foregoing embodiments are only intended to illustrate a preferred mode of the invention but not to limit the scope of the invention. Various variations and modifications made to the technical scheme of the invention by those skilled in the art shall fall within the scope of protection determined by the claims of the invention, without departing from the design spirit of the invention.
Claims (6)
- (1) crushing the plant seeds, soaking the plant seeds in ethanol with the volume concentration of 70% at normal temperature for 3 days, and repeating twice to obtain a leachate;1. A method for extracting oligomeric polyphenolic compounds from plant seeds, characterized by comprising the following steps of:
- 2. The method for extracting the oligomeric polyphenolic compounds from the plant seeds of claim 1, characterized in that the feed-liquid weight ratio of the plant seeds to ethanol in the step (1) is 1: 3-4.(2) distilling the obtained lixivium under reduced pressure to obtain an ethanol extractum;
- 3. The method for extracting the oligomeric polyphenolic compounds from the plant seeds of claim 1, characterized in that the solvent in the step (3) is water, and eluants are gradient eluants of water and ethanol according to ratios, and the mixed eluant is collected from fractions of the gradient elution with water and ethanol according to the ratios (3:7-1:9-0:10).(3) after dissolving the ethanol extractum with a solvent, taking and eluting macroporous resin by a solvent method for decolorization; after concentrating a collected mixed eluant, mixing with silicon for samples, packing, eluting with different eluents respectively, and distilling the eluents under reduced pressure respectively to obtain ethyl acetate extractum and acetone extractum respectively; and (4) taking an acetone extract, separating by high performance liquid chromatography to obtain the oligomeric polyphenol compound, and further to obtain dimeric and trimeric compounds of oligomeric polyphenols when adopting gradient elution with methanol and water according to ratios.
- 4. The method for extracting the oligomeric polyphenolic compounds from the plant seeds of claim 1, characterized in that the eluting process in the step (3) is performed by using 5-8 times of column volume of ethyl acetate, and then performed by using 5-8 times of column volume of acetone, to obtain ethyl acetate extractum and acetone extractum, respectively.2020100587 17 Apr 2020
- 5 the high performance liquid chromatography, under the separation conditions for the componentFl of a ratio of methanol to water being 50:50, a flow rate of 8 mL/min, YMC ODS C18, 250*20 mm, to obtain a compound gentian H; and the component F3 is subjected to high performance liquid chromatography of Waters (a ratio of methanol to water being 45:55, a flow rate of 8 mL/min, YMC ODS Cl8, 250*20 mm) to obtain a compound trans-D-viniferin.5. The method for extracting the oligomeric polyphenolic compounds from the plant seeds of claim 1, characterized in that high performance liquid chromatography in the step (4) is performed, under the conditions of a ratio of methanol to water being 55:45, a flow rate of 8 mL/min, YMC ODS C18, 250*20 mm, to obtain components Fl, F2, F3 and F4, the component Fl is subjected to
- 6. Oligomeric polyphenol compounds prepared by the preparation method of any one of claims 1 to 5, characterized in that the compounds are trans-D-viniferin and trans-gentian H.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114591381A (en) * | 2022-03-23 | 2022-06-07 | 集美大学 | Method for extracting corilagin from fructus Phyllanthi and application thereof |
| CN115010581A (en) * | 2022-06-07 | 2022-09-06 | 新疆维吾尔自治区中药民族药研究所 | Abietane diterpene in hyssop officinalis and preparation method and application thereof |
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2020
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114591381A (en) * | 2022-03-23 | 2022-06-07 | 集美大学 | Method for extracting corilagin from fructus Phyllanthi and application thereof |
| CN115010581A (en) * | 2022-06-07 | 2022-09-06 | 新疆维吾尔自治区中药民族药研究所 | Abietane diterpene in hyssop officinalis and preparation method and application thereof |
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