CN106928287A - A kind of method that phenolic glycoside compound is extracted from lepidii,semen and its application - Google Patents
A kind of method that phenolic glycoside compound is extracted from lepidii,semen and its application Download PDFInfo
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- 210000000582 semen Anatomy 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 22
- 229930182487 phenolic glycoside Natural products 0.000 title claims abstract description 20
- -1 phenolic glycoside compound Chemical class 0.000 title claims abstract description 20
- 229930182470 glycoside Natural products 0.000 claims abstract description 68
- 150000002338 glycosides Chemical class 0.000 claims abstract description 68
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 65
- 239000000284 extract Substances 0.000 claims abstract description 40
- 230000002107 myocardial effect Effects 0.000 claims abstract description 17
- 239000003814 drug Substances 0.000 claims abstract description 7
- 238000004321 preservation Methods 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 282
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 197
- 238000010828 elution Methods 0.000 claims description 46
- 238000004440 column chromatography Methods 0.000 claims description 34
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 34
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 24
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 claims description 17
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 16
- 238000007689 inspection Methods 0.000 claims description 16
- ZRSNZINYAWTAHE-UHFFFAOYSA-N p-methoxybenzaldehyde Chemical compound COC1=CC=C(C=O)C=C1 ZRSNZINYAWTAHE-UHFFFAOYSA-N 0.000 claims description 14
- 238000005507 spraying Methods 0.000 claims description 13
- 239000006228 supernatant Substances 0.000 claims description 10
- AZHSSKPUVBVXLK-UHFFFAOYSA-N ethane-1,1-diol Chemical compound CC(O)O AZHSSKPUVBVXLK-UHFFFAOYSA-N 0.000 claims description 8
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 8
- 238000001556 precipitation Methods 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 6
- 235000004204 Foeniculum vulgare Nutrition 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- QBUKAFSEUHGMMX-MTJSOVHGSA-N (5z)-5-[[3-(1-hydroxyethyl)thiophen-2-yl]methylidene]-10-methoxy-2,2,4-trimethyl-1h-chromeno[3,4-f]quinolin-9-ol Chemical compound C1=CC=2NC(C)(C)C=C(C)C=2C2=C1C=1C(OC)=C(O)C=CC=1O\C2=C/C=1SC=CC=1C(C)O QBUKAFSEUHGMMX-MTJSOVHGSA-N 0.000 claims 2
- 240000006927 Foeniculum vulgare Species 0.000 claims 1
- 230000004083 survival effect Effects 0.000 abstract description 10
- 230000006698 induction Effects 0.000 abstract description 9
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 4
- 238000013459 approach Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 23
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 14
- 239000012071 phase Substances 0.000 description 11
- 241000700159 Rattus Species 0.000 description 9
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 8
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 7
- 238000001228 spectrum Methods 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 5
- 239000008103 glucose Substances 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 241000801118 Lepidium Species 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000003795 desorption Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 239000011347 resin Substances 0.000 description 4
- 229920005989 resin Polymers 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- 241000212314 Foeniculum Species 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 3
- 238000002329 infrared spectrum Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 238000001026 1H--1H correlation spectroscopy Methods 0.000 description 2
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241001657511 Lepidium apetalum Species 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 210000004413 cardiac myocyte Anatomy 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 2
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000001543 one-way ANOVA Methods 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 description 1
- 244000264242 Descurainia sophia Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000010165 autogamy Effects 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000003177 cardiotonic effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000008065 myocardial cell damage Effects 0.000 description 1
- 235000010603 pastilles Nutrition 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/203—Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to from lepidii,semen extract phenolic glycoside compound method and its application, can effectively excavate the new role of lepidii,semen, its solve technical scheme be, the northern new glycosides A of Roripa, molecular formula is C18H24O12[M+Na]+M/z 455.1196, the northern new glycosides B of Roripa, molecular formula is C18H24O12[M+Na]+M/z 455.1202, the present invention is therefrom separated from the water extract of lepidii,semen and identifies two new phenolic glycoside compounds, i.e.,：The northern new glycosides A of the Roripa and new glycosides B of northern Roripa, and two new phenolic glycoside compounds can significantly improve the cell survival rate that the rat myocardial cell H9c2 of hydrogen peroxide induction is damaged, and with myocardial cell protection effect, new approach be provided to prepare myocardial preservation class medicine.
Description
Technical field
The present invention relates to field of medicaments, it is particularly a kind of from lepidii,semen extract phenolic glycoside compound method and its should
With.
Background technology
Lepidium seed is parts of generic medicinal plants, first recorded in《Sheng Nong's herbal classic》, careless subordinate's product are classified as, the flavour of a drug are pungent, hardship, and property is big
It is cold, return lung and bladder warp, have functions that to lead off relieving asthma, line water detumescence.2015 editions《Chinese Pharmacopoeia》" lepidium seed " included has
North and south point, the dry mature seed of wherein crucifer Lepidium apetalum Lepidium apetalum Willd. is practised and claims " north
Lepidium seed ", and the dry mature seed of Descurainia sophia Descurainia sophia (L.) Webbex Prantl. is practised and claims " southern Roripa montana
Son ".Modern pharmacology shows that lepidii,semen mainly has cardiotonic, and its chemical composition is mainly flavonoids.And from Roripa montana
New compound is separated in the water extract of son and the relevant report that its application yet there are no this respect so far is studied.
The content of the invention
For above-mentioned situation, to solve the defect of prior art, the purpose of the present invention is just to provide one kind from lepidii,semen
The middle method for extracting phenolic glycoside compound and its application, can effectively excavate the new role of lepidii,semen.
The present invention solve technical scheme be, from lepidii,semen extract phenolic glycoside compound method：Take the north after processing
Lepidium seed, plus the water of 10 times of weight of lepidii,semen is extracted three times, and each 1.5h, extract solution is concentrated under reduced pressure into medicinal extract shape, concentration
The volumetric concentration of 2-3 times of weight of medicinal extract lepidii,semen is 80% ethanol alcohol precipitation, and supernatant centrifugal filtration is concentrated into without alcohol
Taste, upper DianionHP-20 posts (absorption of highly porous styrene/desorption resin), successively with water, 20% ethanol (v/v),
40% ethanol (v/v), 60% ethanol (v/v), 95% ethanol (v/v) wash-out, by the drying concentrated under reduced pressure of each position, wherein 20% second
Alcohol elution fraction passes through Toyopearl HW-40 column chromatographys, successively with water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 100%
Methanol elution gradient, obtains component A1~A5 and (corresponds to water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 100% methyl alcohol wash-out respectively
It is position, as follows), component A1 through ODS-18 reversed-phase column chromatographies, successively with water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol,
40% methyl alcohol, 100% methanol elution gradient, obtain B1~B6 components, B1 through Sephadex LH-20 column chromatographys, successively with water,
70% methanol elution gradient, (its proportioning is anisaldehyde to the anisaldehyde-concentrated sulfuric acid：The concentrated sulfuric acid：95% ethanol=1:2:100) spray thin
Layer inspection is known, and Sephadex LH-20 posts are gone up in the part for merging anisaldehyde-concentrated sulfuric acid colour developing again, and 100% methyl alcohol is eluted repeatedly, fennel
Fragrant aldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and merges coloured moiety, and component B1 method described above is repeated three times, obtains component C1,
Component C1 again through Toyopearl HW-40 column chromatographys, successively with water, 20% ethanol, 40% ethanol, 70% ethanol gradient elution,
Component D1-D4 is obtained, wherein component D4 prepares HPLC and separates with half, and chromatographic column is YMC-Pack ODS-A chromatographic column (specifications and models：
250 × 10mm, 5 μm of particle diameter, aperture 12nm), mobile phase is acetonitrile:0.05% trifluoroacetic acid (v/v)=10:90, obtain northern Roripa
The new glycosides A and new glycosides B of northern Roripa；
Wherein, the new glycosides A of northern Roripa, molecular formula is C18H24O12[M+Na]+M/z 455.1196 (calculated value 455.1165).Its
Structural formula is as follows：
The northern new glycosides B of Roripa, molecular formula is C18H24O12[M+Na]+M/z 455.1202 (calculated value 455.1165), its structural formula
It is as follows：
The present invention is therefrom separated from the water extract of lepidii,semen and identifies two new phenolic glycoside compounds, i.e.,：Northern Roripa
The new glycosides A and new glycosides B of northern Roripa, and two new phenolic glycoside compounds can significantly improve hydrogen peroxide (H2O2) induction rat myocardial cell
The cell survival rate that H9c2 is damaged, with myocardial cell protection effect, new approach is provided to prepare myocardial preservation class medicine.
Brief description of the drawings
Fig. 1 is the new glycosides A of north Roripa of the invention1H-NMR composes (CD3OD)。
Fig. 2 is the new glycosides A of north Roripa of the invention13C-NMR composes (CD3OD)。
Fig. 3 is that the DEPT 135 of the new glycosides A of north Roripa of the invention composes (CD3OD)。
Fig. 4 is the new glycosides A of north Roripa of the invention1H-1H COSY are composed.
Fig. 5 is the hsqc spectrum of the new glycosides A of north Roripa of the invention.
Fig. 6 is the HMBC spectrums of the new glycosides A of north Roripa of the invention.
Fig. 7 is the NOESY spectrums of the new glycosides A of north Roripa of the invention.
Fig. 8 is the MS spectrums of the new glycosides A of north Roripa of the invention.
Fig. 9 is the IR spectrums of the new glycosides A of north Roripa of the invention.
Figure 10 is the MS spectrums of the new glycosides A of north Roripa of the invention.
Figure 11 is the new glycosides B of north Roripa of the invention1H-NMR composes (CD3OD)。
Figure 12 is the new glycosides B of north Roripa of the invention13C-NMR composes (CD3OD)。
Figure 13 is that the DEPT 135 of the new glycosides B of north Roripa of the invention composes (CD3OD)。
Figure 14 is the new glycosides B of north Roripa of the invention1H-1H COSY are composed.
Figure 15 is the hsqc spectrum of the new glycosides B of north Roripa of the invention.
Figure 16 is the HMBC spectrums of the new glycosides B of north Roripa of the invention.
Figure 17 is the NOESY spectrums of the new glycosides B of north Roripa of the invention.
Figure 18 is the MS spectrums of the new glycosides B of north Roripa of the invention.
Figure 19 is the IR spectrums of the new glycosides B of north Roripa of the invention.
Figure 20 is the MS spectrums of the new glycosides B of north Roripa of the invention.
Figure 21 is the present invention north new glycosides A of Roripa to H2O2The H9c2 rat myocardial cell survival rates of induction influence (N=
6)。
Figure 22 is the present invention north new glycosides B of Roripa to H2O2The H9c2 rat myocardial cell survival rates of induction influence (N=
6)。
Specific embodiment
Specific embodiment of the invention is described in further detail with reference to embodiments.
Embodiment 1
Lepidii,semen 8kg is taken, in 240 DEG C, 5.5min is fried, plus the water of 10 times of weight of lepidii,semen extracts three times, every time
1.5h, extract solution is concentrated under reduced pressure into medicinal extract, the medicinal extract ethanol alcohol precipitation that 20L volumetric concentrations are 80% of concentration, by supernatant from
The heart is filtered, and is concentrated into without alcohol taste, and upper Dianion HP-20 posts (absorption of highly porous styrene/desorption resin) are used successively
Water, 20% ethanol (v/v), 40% ethanol (v/v), 60% ethanol (v/v), 95% ethanol (v/v) wash-out, depressurize dense by each position
Contracting drying, respectively obtain water, 20% ethanol, 40% ethanol, 60% ethanol, 95% ethanol elution (respectively 361.0g, 71.2g,
89.6g, 67.2g, 28.7g) component, wherein 20% ethanol elution component by Toyopearl HW-40 column chromatographys, is used successively
Water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 100% methanol elution gradient, obtain component A1~A5, component A1 (25.3g) warps
ODS-18 reversed-phase column chromatographies, are washed with water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 40% methyl alcohol, 100% methanol gradient successively
It is de-, obtain B1~B6, component B1 (7.8g) through Sephadex LH-20 column chromatographys, successively with water, 70% methanol elution gradient, fennel
Fragrant aldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and anisaldehyde-concentrated sulfuric acid presses anisaldehyde：The concentrated sulfuric acid：95% ethanol=1:2:100 proportionings, close
And Sephadex LH-20 posts are gone up in the part of anisaldehyde-concentrated sulfuric acid colour developing again, 100% methyl alcohol is eluted repeatedly, anisaldehyde-dense sulphur
Acid spraying thin layer inspection is known, and merges coloured moiety, and component B1 method described above is repeated three times, obtains component C1 (1.2g), group
Divide C1 again through Toyopearl HW-40 column chromatographys, successively with water, 20% ethanol, 40% ethanol, 70% ethanol gradient elution, obtain
Component D1-D4, wherein component D4 (50.3mg) prepare HPLC and separate with half, and chromatographic column is YMC-Pack ODS-A chromatographic columns (rule
Lattice number：250 × 10mm, 5 μm of particle diameter, aperture 12nm), mobile phase is the trifluoroacetic acid of acetonitrile -0.05% (volume ratio 10:90),
Obtain new glycosides A (14.0mg, the t of northern RoripaR=80.3min) and new glycosides B (2.9mg, the t of northern RoripaR=62.5min).
Embodiment 2
Lepidii,semen 2kg is taken, in 240 DEG C, 5.5min is fried, plus the water of 10 times of weight of lepidii,semen extracts three times, every time
1.5h, extract solution is concentrated under reduced pressure into medicinal extract, and supernatant is centrifuged the medicinal extract ethanol alcohol precipitation that 5L volumetric concentrations are 80% of concentration
Filtering, be concentrated into without alcohol taste, upper Dianion HP-20 posts (absorption of highly porous styrene/desorption resin), successively with water,
20% ethanol (v/v), 40% ethanol (v/v), 60% ethanol (v/v), 95% ethanol (v/v) wash-out, each position is concentrated under reduced pressure
Dry, respectively obtain water, 20% ethanol, 40% ethanol, 60% ethanol, 95% ethanol elution (respectively 90.1g, 18.0g,
22.5g, 17.0g, 7.3g) component, wherein 20% ethanol elution component by Toyopearl HW-40 column chromatographys, is used successively
Water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 100% methanol elution gradient, obtain component A1~A5, component A1 (6.2g) warps
ODS-18 reversed-phase column chromatographies, are washed with water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 40% methyl alcohol, 100% methanol gradient successively
It is de-, obtain B1~B6, component B1 (1.9g) through Sephadex LH-20 column chromatographys, successively with water, 70% methanol elution gradient, fennel
Fragrant aldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and anisaldehyde-concentrated sulfuric acid presses anisaldehyde：The concentrated sulfuric acid：95% ethanol=1:2:100 proportionings, close
And Sephadex LH-20 posts are gone up in the part of anisaldehyde-concentrated sulfuric acid colour developing again, 100% methyl alcohol is eluted repeatedly, anisaldehyde-dense sulphur
Acid spraying thin layer inspection is known, and merges coloured moiety, and component B1 method described above is repeated three times, obtains component C1 (300mg), group
Divide C1 again through Toyopearl HW-40 column chromatographys, successively with water, 20% ethanol, 40% ethanol, 70% ethanol gradient elution, obtain
Component D1-D4, wherein component D4 (13.0mg) prepare HPLC and separate with half, and chromatographic column is YMC-Pack ODS-A chromatographic columns (rule
Lattice number：250 × 10mm, 5 μm of particle diameter, aperture 12nm), mobile phase is the trifluoroacetic acid of acetonitrile -0.05% (10:90) north, is obtained
New glycosides A (3.2mg, the t of RoripaR=80.3min) and new glycosides B (0.6mg, the t of northern RoripaR=62.5min).
Embodiment 3
Lepidii,semen 1kg is taken, in 240 DEG C, 5.5min is fried, plus the water of 10 times of weight of lepidii,semen extracts three times, every time
1.5h, extract solution is concentrated under reduced pressure into medicinal extract, and supernatant is centrifuged the medicinal extract ethanol alcohol precipitation that 3L volumetric concentrations are 80% of concentration
Filtering, be concentrated into without alcohol taste, upper Dianion HP-20 posts (absorption of highly porous styrene/desorption resin), successively with water,
20% ethanol (v/v), 40% ethanol (v/v), 60% ethanol (v/v), 95% ethanol (v/v) wash-out, each position is concentrated under reduced pressure
Dry, respectively obtain water, 20% ethanol, 40% ethanol, 60% ethanol, 95% ethanol elution (respectively 45.0g, 9.0g,
11.3g, 8.5g, 3.6g) component, wherein 20% ethanol elution component (9.0g) is by Toyopearl HW-40 column chromatographys, according to
Secondary use water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 100% methanol elution gradient, obtain component A1~A5, component A1 (3.2g)
Through ODS-18 reversed-phase column chromatographies, successively with water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 40% methyl alcohol, 100% methanol gradient
Wash-out, obtain B1~B6, component B1 (0.9g) through Sephadex LH-20 column chromatographys, successively with water, 70% methanol elution gradient,
Anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and anisaldehyde-concentrated sulfuric acid presses anisaldehyde：The concentrated sulfuric acid：95% ethanol=1:2:100 proportionings,
Sephadex LH-20 posts are gone up in the part for merging anisaldehyde-concentrated sulfuric acid colour developing again, and 100% methyl alcohol is eluted repeatedly, anisaldehyde-dense
The inspection of sulfate spray thin layer is known, and merges coloured moiety, and component B1 method described above is repeated three times, obtains component C1 (150mg)
Component, C1 again through Toyopearl HW-40 column chromatographys, successively with water, 20% ethanol, 40% ethanol, 70% ethanol gradient elution,
Component D1-D4 is obtained, wherein component D4 (6.3mg) prepares HPLC and separates with half, chromatographic column is YMC-Pack ODS-A chromatographic columns (rule
Lattice number：250 × 10mm, 5 μm of particle diameter, aperture 12nm), mobile phase is the trifluoroacetic acid of acetonitrile -0.05% (10:90) north, is obtained
New glycosides A (1.8mg, the t of RoripaR=80.0min) and new glycosides B (0.4mg, the t of northern RoripaR=62.0min).
For the effective substance of clear and definite lepidii,semen, the present invention is extracted using decocting cooking method to it, and due to
It meets water stickness so that decocting liquid recovery rate is low not to be said, the composition for extracting mostly mucilaginous substance, therefore the present invention is first
First using method is fried, in 240 DEG C, frying 5.5min is simply processed to it, so that the composition in lepidii,semen is more easy to dissolution,
Two new phenolic glycoside compounds are therefrom separated and identified from the water extract of lepidii,semen, i.e.,：The northern new glycosides A of Roripa and the new glycosides of northern Roripa
B, experimental result shows that the two new phenolic glycoside compounds can significantly improve the rat myocardial cell of hydrogen peroxide (H2O2) induction
The cell survival rate that H9c2 is damaged, with myocardial cell protection effect.And through experiment fully prove, correlation test data
It is as follows：
1 instrument and reagent
Nuclear magnetic resonance is used with the NMRs of BrukerAVANCE III 500 (TMS internal standards) (Bruker), infrared spectrum
The Microscope Spectrometer of Nicolet is 10 (Thermo Scientific, USA), high resolution mass spectrum is used
Bruker maxis HD mass spectrometer, ultraviolet spectra Shimadzu UV-2401PC apparatus, efficiently
Phase chromatography-use Waters Alliance 2695 efficient liquid phase systems of series, are equipped with 2998 type PDADs,
Empower3 Data Processing in Chromatography Workstation, LC50 type high pressure preparative liquid chromatographs, UV200 types UV-detector [the sharp think of of match spectrum (north
Capital) Science and Technology Ltd.], YMC-Pack ODS-A chromatographic columns (250 × 10mm.D.S-5mm, 12mm) (YMC Co., Ltds), its
It is remaining to have N-1100 types Rotary Evaporators (Shanghai Ai Lang Instrument Ltd), A-1000S type current air exhauster (Shanghai Ai Lang instruments
Co., Ltd), N-1111 types freeze water circle device (Shanghai Ai Lang Instrument Ltd), FDU-2110 type freeze driers
(Shanghai Ai Lang Instrument Ltd), DFZ-60508 types vacuum drying chamber (Shanghai Yiheng Scientific Instruments Co., Ltd), AB204-
N a ten thousandths analytical precision balances (METTLER TOLEDO), iMARK types ELIASA (U.S. BIO-RAD), carbon dioxide culture
Case (Shanghai STIK), superclean bench (Su Jing groups), the super combined Superpure water machines (SARTORIUS) of Arium 611VF,
Put microscope (Nikon), PB-10 acidometers (German Sai Duolisi groups).
Column chromatography filler Diaion HP-20, MCI Gel CHP-20 (Mitsubishi chemical company), Toyopearl HW-
40 (Japanese TOSOH companies), Sephadex LH-20 (Parmacia Biotech companies), column chromatography used silica gel H (160-
200 mesh) it is Haiyang Chemical Plant, Qingdao's production, culture dish, 96 well culture plates, cell cryopreservation tube (Corning companies), chromatogram used
The production of pure reagent position α Cygni friend's fine chemicals Co., Ltd, AR used is that Beijing Chemical Plant and Tianjin the 3rd are changed
Learn chemical reagent work's production.
DMEM sugared nutrient solutions (Gibco) high, hyclone (Zhejiang Tian Hang bio tech ltd), trypsase
(Gibco), DMSO (Solarbio)；MTT (Biosharp), H2O2Solution (Tianjin Heng Xing chemical reagent Manufacturing Co., Ltd),
Water is ultra-pure water, and PBS etc. is autogamy.
Lepidii,semen picked up from Nanyang, henan in 2014, was identified as crucifer Lepidium apetalum Lepidium
The dry mature seed of apetalum Willd..
2 Structural Identifications
The northern new glycosides A of Roripa, light yellow crystalline powder (MeOH), molecular formula is C18H24O12[M+Na]+m/z 455.1196
(calculated value 455.1165), UV (MeOH) λmax:206.0,230.0,315.0nm；IR(KBr)νmaxcm-1：3364,2931,
1576,1489,1039.Its structural formula and nuclear magnetic data are as follows：
The northern new glycosides B of Roripa, light yellow crystalline powder (MeOH), molecular formula is C18H24O12[M+Na]+m/z 455.1202
(calculated value 455.1165), UV (MeOH) λmax:206.0,238.0,302.0nm；IR(KBr)νmaxcm-1：3413,2925,
1672,1466,1246,1046.Its structural formula and nuclear magnetic data are as follows：
The new glycosides A's and B of the northern Roripa of table 11H NMR (500MHz) and13C NMR (125MHz) data (in CD3OD)
The 3 north new glycosides A of Roripa and the north new glycosides B of Roripa are to hydrogen peroxide (H2O2) injury rats cardiac muscle cell H9c2 protective effect experiment
3.1 cells
H9c2 cell lines are purchased from Shenzhen Biowit Biotechnologies Co., Ltd..
3.2 cell culture
H9c2 cultures (are contained into penicillin and streptomysin in being the DMEM culture mediums of 10% hyclone containing volume fraction
100kU·L-1), it is placed in 37 DEG C, 5%CO2Cultivated in saturated humidity incubator, growth period cell of taking the logarithm is used to test.
3.3H2O2The foundation of the H9c2 rat myocardial cell damage models of induction
During Secondary Culture, by the cell of exponential phase with 5 × 104Density be inoculated in 96 orifice plates, treat that cell is complete
After adherent, the DMEM in high glucose nutrient solution of pastille is used instead.H2O2Give containing 200 μM of H2O2The nutrient solution culture of DMEM in high glucose containing serum
6h。
3.4 experiment packets
Every group sets 6 parallel holes, if normal group (NC), H2O2Each monomeric compound administration group of group (M), lepidii,semen.H2O2
Group：Give containing 200 μM of H2O2The nutrient solution culture of DMEM in high glucose containing serum 6h；The northern new glycosides A of Roripa and the new glycosides B administration groups of northern Roripa：Point
Do not give containing 200 μM of H2O2With the culture of DMEM in high glucose containing serum of 1,2,4,8,16,32,64 μM of north new glycosides A of Roripa and the new glycosides B of northern Roripa
Liquid culture 6h.
3.5MTT methods detect the north new glycosides A of Roripa and influences of the north new glycosides B of Roripa to H9c2 damaging cells survival rates
Cell is inoculated in 96 orifice plates, after adherent rear dosing intervention culture, absorbs supernatant, and each hole is separately added into containing 1%MTT
DMEM in high glucose nutrient solution 200 μ l, 37 DEG C of incubation 4h of (5g/L), absorb supernatant, add the DMSO of 150 μ L, concussion 10min to make
Crystallization fully dissolving.OD values at 490nm wavelength are detected with ELIASA, cell survival rate=(OD experimental groups/OD pairs are calculated with formula
According to group) × 100%.Experiment is repeated 3 times, and averages.
3.6 statistical methods
Experimental data mean ± standard deviationRepresent, with SPSS18.0 statistical softwares processing data, compare between each group
Using one-way analysis of variance (One-WayANOVA), P<0.05 indicates significant, P<0.01 indicates pole conspicuousness
Meaning.3.7 results
Result of study shows：Compared with blank group, model group cell viability significantly reduces (P<0.01)；With model group phase
When than, northern new glycosides A dosages of Roripa being 1 μM, can pole significantly improve cell viability (P<0.01), at 2,4,8,16 μM, energy
Enough significantly improve cell viability (P<0.05)；The northern new glycosides B dosages of Roripa can significantly improve cell at 4,8,16,32 μM
Vigor (P<0.05).Show that the lepidii,semen monomer north new glycosides A and B of Roripa has certain protective effect to cardiac muscle cell.
The north of the table 2 new glycosides A of Roripa is to H2O2The H9c2 rat myocardial cell survival rates of induction influence (N=6)
Note：Compare with normal group:*P<0.05；**P<0.01 and H2O2Group compares:#P<0.05；##P<0.01
The north of the table 3 new glycosides B of Roripa is to H2O2The H9c2 rat myocardial cell survival rates of induction influence (N=6)
Note：Compare with normal group:*P<0.05；**P<0.01 and H2O2Group compares:#P<0.05；##P<0.01
To sum up, separated from the water extract of lepidii,semen and two new phenolic glycoside compounds identifying, experimental result shows
The two new phenolic glycoside compounds can significantly improve hydrogen peroxide (H2O2) induction rat myocardial cell H9c2 damage cell survival
Rate, with myocardial cell protection effect, can be effectively used for preparing myocardial preservation class medicine, develop the new application of lepidii,semen,
With actual clinical meaning and good application and popularization value.
Claims (7)
1. it is a kind of from lepidii,semen extract phenolic glycoside compound, it is characterised in that the northern new glycosides A of Roripa, molecular formula is C18H24O12[M
+Na]+M/z 455.1196, its structural formula is as follows：
The northern new glycosides B of Roripa, molecular formula is C18H24O12[M+Na]+M/z 455.1202, its structural formula is as follows：
2. described in claim 1 from lepidii,semen extract phenolic glycoside compound extracting method, it is characterised in that take processing
Lepidii,semen afterwards, plus the water of 10 times of weight of lepidii,semen is extracted three times, and each 1.5h, extract solution is concentrated under reduced pressure into medicinal extract shape,
The volumetric concentration of 2-3 times of weight of medicinal extract lepidii,semen of concentration is 80% ethanol alcohol precipitation, by supernatant centrifugal filtration, concentration
Extremely without alcohol taste, upper Dianion HP-20 posts, successively with water, 20% ethanol, 40% ethanol, 60% ethanol, 95% ethanol elution,
Each position is concentrated under reduced pressure drying, wherein 20% ethanol elution component is by Toyopearl HW-40 column chromatographys, successively with water,
10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 100% methanol elution gradient, obtain component A1~A5, and component A1 is anti-phase through ODS-18
Column chromatography, successively with water, 10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 40% methyl alcohol, 100% methanol elution gradient, obtains B1
, through Sephadex LH-20 column chromatographys, successively with water, 70% methanol elution gradient, anisaldehyde-concentrated sulfuric acid is sprayed for~B6, component B1
Thin layer inspection is known, and Sephadex LH-20 posts are gone up in the part for merging anisaldehyde-concentrated sulfuric acid colour developing again, and 100% methyl alcohol is eluted repeatedly,
Anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and merges coloured moiety, and component B1 method described above is repeated three times, obtains component
C1, component C1 through Toyopearl HW-40 column chromatographys, are washed with water, 20% ethanol, 40% ethanol, 70% ethanol gradient successively again
It is de-, component D1-D4 is obtained, wherein component D4 prepares HPLC and separates with half, and chromatographic column is YMC-Pack ODS-A chromatographic columns, mobile phase
It is acetonitrile:0.05% trifluoroacetic acid=10:90, obtain the new glycosides A of the northern Roripa and new glycosides B of northern Roripa.
3. it is according to claim 2 from lepidii,semen extract phenolic glycoside compound extracting method, it is characterised in that take
Lepidii,semen 8kg, in 240 DEG C, fries 5.5min, plus the water of 10 times of weight of lepidii,semen is extracted three times, each 1.5h, extract solution
It is concentrated under reduced pressure into medicinal extract, the medicinal extract ethanol alcohol precipitation that 20L volumetric concentrations are 80% of concentration, by supernatant centrifugal filtration, concentration
Extremely without alcohol taste, upper Dianion HP-20 posts, successively with water, 20% ethanol, 40% ethanol, 60% ethanol, 95% ethanol elution,
By the drying concentrated under reduced pressure of each position, water, 20% ethanol, 40% ethanol, 60% ethanol, the group of 95% ethanol elution are respectively obtained
Point, wherein 20% ethanol elution component is by Toyopearl HW-40 column chromatographys, successively with water, 10% methyl alcohol, 20% methyl alcohol,
30% methyl alcohol, 100% methanol elution gradient, obtain component A1~A5, component A1 through ODS-18 reversed-phase column chromatographies, successively with water,
10% methyl alcohol, 20% methyl alcohol, 30% methyl alcohol, 40% methyl alcohol, 100% methanol elution gradient, obtain B1~B6, component B1 warps
Sephadex LH-20 column chromatographys, successively with water, 70% methanol elution gradient, anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, fennel
Aldehyde-concentrated sulfuric acid presses anisaldehyde：The concentrated sulfuric acid：95% ethanol=1:2:100 proportionings, merge the part of anisaldehyde-concentrated sulfuric acid colour developing again
Secondary upper Sephadex LH-20 posts, 100% methyl alcohol is eluted repeatedly, and anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and merges colour developing portion
Point, component B1 method described above is repeated three times, obtains component C1, component C1 again through Toyopearl HW-40 column chromatographys, according to
Secondary use water, 20% ethanol, 40% ethanol, 70% ethanol gradient elution, obtain component D1-D4, and wherein component D4 prepares HPLC with half
Separate, chromatographic column is YMC-Pack ODS-A chromatographic columns, mobile phase is trifluoroacetic acid=10 of acetonitrile -0.05%:90, obtain northern Roripa
The new glycosides A and new glycosides B of northern Roripa.
4. it is according to claim 2 from lepidii,semen extract phenolic glycoside compound extracting method, it is characterised in that take
Lepidii,semen 2kg, in 240 DEG C, fries 5.5min, plus the water of 10 times of weight of lepidii,semen is extracted three times, each 1.5h, extract solution
Medicinal extract is concentrated under reduced pressure into, the medicinal extract ethanol alcohol precipitation that 5L volumetric concentrations are 80% of concentration, by supernatant centrifugal filtration, is concentrated into
Without alcohol taste, upper Dianion HP-20 posts, successively with water, 20% ethanol, 40% ethanol, 60% ethanol, 95% ethanol elution, will
Each position is concentrated under reduced pressure drying, respectively obtains water, 20% ethanol, 40% ethanol, 60% ethanol, the component of 95% ethanol elution,
Wherein 20% ethanol elution component passes through Toyopearl HW-40 column chromatographys, successively with water, 10% methyl alcohol, 20% methyl alcohol, 30%
Methyl alcohol, 100% methanol elution gradient, obtain component A1~A5.Component A1 through ODS-18 reversed-phase column chromatographies, successively with water, 10% first
Alcohol, 20% methyl alcohol, 30% methyl alcohol, 40% methyl alcohol, 100% methanol elution gradient, obtain B1~B6, and component B1 is through Sephadex
LH-20 column chromatographys, successively with water, 70% methanol elution gradient, anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, the anisaldehyde-concentrated sulfuric acid
By anisaldehyde：The concentrated sulfuric acid：95% ethanol=1:2:Go up again 100 proportionings, the part for merging anisaldehyde-concentrated sulfuric acid colour developing
Sephadex LH-20 posts, 100% methyl alcohol is eluted repeatedly, and anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and merges coloured moiety, group
Divide B1 method described above to be repeated three times, obtain component C1, component C1 through Toyopearl HW-40 column chromatographys, is used successively again
Water, 20% ethanol, 40% ethanol, 70% ethanol gradient elution, obtain component D1-D4, and wherein component D4 prepares HPLC and separates with half,
Chromatographic column is YMC-Pack ODS-A chromatographic columns, and mobile phase is trifluoroacetic acid=10 of acetonitrile -0.05%:90, obtain the new glycosides A of northern Roripa
Glycosides B new with northern Roripa.
5. it is according to claim 2 from lepidii,semen extract phenolic glycoside compound extracting method, it is characterised in that take
Lepidii,semen 1kg, in 240 DEG C, fries 5.5min, plus the water of 10 times of weight of lepidii,semen is extracted three times, each 1.5h, extract solution
Medicinal extract is concentrated under reduced pressure into, the medicinal extract ethanol alcohol precipitation that 3L volumetric concentrations are 80% of concentration, by supernatant centrifugal filtration, is concentrated into
Without alcohol taste, upper Dianion HP-20 posts, successively with water, 20% ethanol, 40% ethanol, 60% ethanol, 95% ethanol elution, will
Each position is concentrated under reduced pressure drying, respectively obtains water, 20% ethanol, 40% ethanol, 60% ethanol, the component of 95% ethanol elution,
Wherein 20% ethanol elution component passes through Toyopearl HW-40 column chromatographys, successively with water, 10% methyl alcohol, 20% methyl alcohol, 30%
Methyl alcohol, 100% methanol elution gradient, obtain component A1~A5, component A1 through ODS-18 reversed-phase column chromatographies, successively with water, 10% first
Alcohol, 20% methyl alcohol, 30% methyl alcohol, 40% methyl alcohol, 100% methanol elution gradient, obtain B1~B6, and component B1 is through Sephadex
LH-20 column chromatographys, successively with water, 70% methanol elution gradient, anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, the anisaldehyde-concentrated sulfuric acid
By anisaldehyde：The concentrated sulfuric acid：95% ethanol=1:2:Go up again 100 proportionings, the part for merging anisaldehyde-concentrated sulfuric acid colour developing
Sephadex LH-20 posts, 100% methyl alcohol is eluted repeatedly, and anisaldehyde-concentrated sulfuric acid spraying thin layer inspection is known, and merges coloured moiety, group
Divide B1 method described above to be repeated three times, obtain component C1 components C1 again through Toyopearl HW-40 column chromatographys, use successively
Water, 20% ethanol, 40% ethanol, 70% ethanol gradient elution, obtain component D1-D4, and wherein component D4 prepares HPLC and separates with half,
Chromatographic column is YMC-Pack ODS-A chromatographic columns, and mobile phase is trifluoroacetic acid=10 of acetonitrile -0.05%:90, obtain the new glycosides A of northern Roripa
Glycosides B new with northern Roripa.
6. application of the phenolic glycoside compound described in claim 1 in myocardial preservation medicine is prepared.
7. the new glycosides A of northern Roripa, the north new glycosides B of Roripa that extracting method described in any one of claim 2-5 is extracted are preparing myocardial preservation medicine
Application in thing.
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| CN106866753A (en) * | 2017-03-02 | 2017-06-20 | 河南中医药大学 | A kind of method that sulphur glycoside compound is extracted from lepidii,semen and its application |
| CN107353315A (en) * | 2017-07-29 | 2017-11-17 | 河南中医药大学 | A kind of method and its application that the new alkali A compounds of northern Roripa are extracted from lepidii,semen |
| CN107446008A (en) * | 2017-09-06 | 2017-12-08 | 河南中医药大学 | A kind of new glycosides E of northern Roripa and preparation method and application |
| CN107602640A (en) * | 2017-10-18 | 2018-01-19 | 河南中医药大学 | A kind of method and its application that northern Roripa neoflavone C is extracted from lepidii,semen |
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| CN108341847A (en) * | 2017-11-29 | 2018-07-31 | 河南中医药大学 | The extraction north new alkali G of Roripa and the north new alkali H of Roripa and the preparation method and application thereof in a kind of lepidii,semen |
| CN115671154A (en) * | 2022-11-29 | 2023-02-03 | 浙江大学 | Semen Lepidii flavone component, its extraction method and application in preparing medicine for treating or preventing heart failure |
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| CN106866753A (en) * | 2017-03-02 | 2017-06-20 | 河南中医药大学 | A kind of method that sulphur glycoside compound is extracted from lepidii,semen and its application |
| CN107353315A (en) * | 2017-07-29 | 2017-11-17 | 河南中医药大学 | A kind of method and its application that the new alkali A compounds of northern Roripa are extracted from lepidii,semen |
| CN107446008A (en) * | 2017-09-06 | 2017-12-08 | 河南中医药大学 | A kind of new glycosides E of northern Roripa and preparation method and application |
| CN107446008B (en) * | 2017-09-06 | 2020-10-02 | 河南中医药大学 | Neoside E, and preparation method and application thereof |
| CN107602640A (en) * | 2017-10-18 | 2018-01-19 | 河南中医药大学 | A kind of method and its application that northern Roripa neoflavone C is extracted from lepidii,semen |
| CN107602639A (en) * | 2017-10-18 | 2018-01-19 | 河南中医药大学 | A kind of method and its application that sesquiterpenoids is extracted from lepidii,semen |
| CN108341847A (en) * | 2017-11-29 | 2018-07-31 | 河南中医药大学 | The extraction north new alkali G of Roripa and the north new alkali H of Roripa and the preparation method and application thereof in a kind of lepidii,semen |
| CN108191936A (en) * | 2018-01-12 | 2018-06-22 | 河南中医药大学 | A kind of new alkali J of northern Roripa extracted from lepidii,semen and the new alkali K of northern Roripa and preparation method and application |
| CN115671154A (en) * | 2022-11-29 | 2023-02-03 | 浙江大学 | Semen Lepidii flavone component, its extraction method and application in preparing medicine for treating or preventing heart failure |
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