CN105820147B - The preparation method and its usage of alpine yallow herb active component - Google Patents

The preparation method and its usage of alpine yallow herb active component Download PDF

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CN105820147B
CN105820147B CN201610281770.7A CN201610281770A CN105820147B CN 105820147 B CN105820147 B CN 105820147B CN 201610281770 A CN201610281770 A CN 201610281770A CN 105820147 B CN105820147 B CN 105820147B
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alpine yallow
yallow herb
active component
water
extract
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CN105820147A (en
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顾政
顾政一
贺金华
戎晓娟
刘君琳
茹仙古丽·依明
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INST OF PHARMACOLOGY XINJIANG UYGUR AUTONOMOUS REGIONS
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    • C07—ORGANIC CHEMISTRY
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    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
    • C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
    • C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
    • C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
    • 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
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    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
    • C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
    • C07D311/40—Separation, e.g. from natural material; Purification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/42—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms in positions 2 and 4
    • C07D311/44—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms in positions 2 and 4 with one hydrogen atom in position 3
    • C07D311/54—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms in positions 2 and 4 with one hydrogen atom in position 3 substituted in the carbocyclic ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea

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Abstract

The present invention relates to a kind of preparation method and its usage of alpine yallow herb active component.This method is by dry alpine yallow herb pulverizing medicinal materials, is extracted through ethanol water, then through polyamide column separating purification, elution position is extracted with organic solvent, the sample that collection chloroform layer obtains carries out secondary separation with silicagel column and purified, and recrystallizes, that is, obtains 3 monomeric compounds, yield and content are respectively casticin 0.30mg/g, 96% 98%, the mg/g of 6 de-methoxy, 4 ' O methyl capillarisin 0.35,97% 99%, the mg/g of artemetine 0.06,95% 97%.Experiment through external resisiting influenza virus shows：There is the alpine yallow herb active component obtained by the method for the invention certain anti-A/ Chinese to prevent/359/95 (H3N2) influenza activity, be potential new resisiting influenza virus lead compound.The method of the invention obtains casticin, the O methyl capillarisin of 6 de-methoxy 4 ', artemetine from alpine yallow herb simultaneously, its low cost, easy, efficiently, stably.

Description

The preparation method and its usage of alpine yallow herb active component
Technical field
The present invention relates to the preparation of alpine yallow herb anti-influenza virus activity composition and purposes.
Background technology：
Artemisia rupestris (Artemisia rupestris L.) is composite family (Compositae) artemisia (ArtemisiaL.) Plant, is Xinjiang native drug, Uygur medicine name-diligent partner Ai Mini.Being distributed mainly on the ground such as Xinjiang, the Central Asia, Europe also has point Cloth.Xinjiang is mainly distributed the Subalpine region grass for along the ground such as the Mountain Ranges of Tian Shan Mountains and Montes Altai, being typically grown in 1600~3000m of height above sea level Former, coniferous forest open ground.Artemisia rupestris is cold in nature, and acrid flavour is bitter, has the functions such as heat clearing away, anti-inflammatory analgetic, removing pattogenic heat from the blood and toxic material from the body.Newly Boundary alpine yallow herb has higher medical value, among the people to use all herbal medicine more, has antiallergy, anti-oxidant, antiviral, liver protection drop Enzyme, cancer cell specific induction of apoptosis, many effects of immunological regulation.In document report alpine yallow herb herb containing flavonoids, Mek-Tol Unit, The chemical compositions such as sesquiterpenoids, amino acids, glycoside, polysaccharide, volatile oil, polypeptide and alkaloid.Early-stage Study finds one Branch parthenium extract Mek-Tol Unit and flavonoids have stronger anti-influenza virus activity, and we compare 26 Artemisia rupestris early stage Different extract anti-influenza A virus effects, as a result find the inhibiting rate of the extract infected by influenza of 4 Artemisia rupestris Reach more than 50%；Find Herba Achilleae extract Mek-Tol Unit and flavonoids to Respiratory Syncytial Virus(RSV) (RSV) and sidestream simultaneously The interaction in vitro of Influenza Virus (PIV) is clearly.Main chemical compositions have in prompting Artemisia rupestris antivirus effective position Potential anti-influenza virus activity.
Artemisia rupestris pharmacological action is extensive, and clinical value is just increasingly paid attention to.It is susceptible for alpine yallow herb anti-current The research of toxic action is more deep, but because monomer component is complicated, synthesis difficulty is big, and access approaches are few, therefore to its pharmacology The research of activity focuses mostly on active component, and the research to the pharmacological activity and its mechanism of action of monomer composition is not still deep Enter, currently there is no the research that pharmacological activity is carried out to the other monomers composition in alpine yallow herb in addition to rupestonic acid.
The content of the invention：
It is an object of the present invention to provide the preparation method and its usage of alpine yallow herb active component.This method is by drying Alpine yallow herb pulverizing medicinal materials, extracted through ethanol water, then through polyamide column separating purification, will elution position with organic molten Agent extracts, and the sample that collection chloroform layer obtains carries out secondary separation with silicagel column and purified, and recrystallizes, that is, 3 lists obtained Body compound, yield are respectively casticin 0.30mg/g, 6- de-methoxy -4 '-O- methyl capillarisin 0.35mg/g, Artemetine 0.06mg/g, content are respectively casticin 96%-98%, 6- de-methoxy -4 '-O- methyl capillarisins 97%-99%, artemetine 95%-97%.Experiment through external resisiting influenza virus shows：Obtained by the method for the invention Alpine yallow herb active component has anti-influenza virus activity, and the infected by influenza A/ Chinese prevents that/359/95 (H3N2) has certain activity, It is potential new resisiting influenza virus lead compound.The method of the invention is inexpensive, easy, efficient, stabilization from one Casticin, 6- de-methoxy -4 '-O- methyl capillarisin, artemetine are extracted in wormwood artemisia.
A kind of preparation method of alpine yallow herb active component of the present invention, follows these steps to carry out：
A. dry alpine yallow herb medicinal material is taken, through crushing, 10 mesh sieves is crossed, is placed in round-bottomed flask, 8-15 times of weight of addition Volumetric concentration is that 30-75% ethanol waters are extracted, and 50-80 DEG C of water bath heating temperature, refluxing extraction 2-5 times, the time is 1-3 hours/time, filtrate after extraction is 1.0-1.5 through being concentrated under reduced pressure into relative density, and it is standby to obtain medicinal extract；
B. the medicinal extract that will be obtained in step a, vacuum dried case are dried, vacuum -0.08MPa, 45-70 DEG C of temperature, obtained It is standby to Herba Achilleae extract；
C. the extract and polyamide step b obtained is nylon-6 or nylon -66 or nylon-610 resin by weight Measure ratio 1:6-1:10 are adsorbed, and resin column blade diameter length ratio is 1:3-1:7, sample solution extract concentrations are 4%-10%, loading stream Speed is 0.5-3BV/h, is first eluted with water polyamide 5-10BV, elution flow rate 0.5-3BV/h after absorption, then by water elution After discarding, the ethanol water for being 10-95% with volume by volume concentration elutes 5-10BV, flow velocity 0.5-3BV/h, collects concentration and is The ethanol eluate of 30-60% gradients, rotated evaporimeter concentration, then vacuum dried case dry, pulverize, and it is pure to produce alpine yallow herb Compound；
D. the alpine yallow herb purified that step c is obtained is suspended with water, petroleum ether layer is discarded with petroleum ether extraction, water layer is with three Chloromethanes equal-volume extraction, chloroform extraction liquid is collected, it is 1.0-1.2 that rotated evaporimeter, which is concentrated into relative density, is obtained Medicinal extract is standby；
E. alpine yallow herb purified medicinal extract step d obtained is with silica gel with weight than 1:1-1:3 are adsorbed and are volatilized, silicon Glue post blade diameter length ratio is 1:5-1:10, wet method is splined on silicagel column, successively with volume ratio 10:0-0:10 petroleum ether:Ethyl acetate Gradient elution, elution flow rate 0.5-3BV/h, thin-layer chromatography monitoring collection liquid, allied substances are merged, respectively rotated evaporimeter It is 1.0-1.5 to be concentrated into relative density, and pressure is -0.08MPa, and temperature is 40 DEG C -60 DEG C, and solvent normal temperature volatilizes, and adds methanol Dissolving, normal temperature evaporate into precipitation crystal, filter, obtain monomeric compound, be respectively that casticin content exists through Structural Identification Between 96%-98%, between 97%-99%, artemetine content exists 6- de-methoxy -4 '-O- methyl capillarisin contents Between 95%-97%.
The alpine yallow herb active component that described method obtains is preparing the prevention and treatment resisiting influenza virus A/ Chinese anti-/ 359/95 (H3N2) the purposes in medicine.
The alpine yallow herb active component that described method obtains prevents/359/95 (H3N2) guarantor preparing the resisiting influenza virus A/ Chinese Purposes in strong product.
Casticin, 6- de-methoxies -4 '-O- methyl capillarisin and the artemetine that the method for the invention obtains It is the lead compound of potential antiviral drugs with antiviral activity.
The preparation method and its usage of alpine yallow herb active component of the present invention, the alpine yallow herb obtained by this method are lived Property composition, it is respectively through Structural Identification：
Casticin, structural formula are：
6- de-methoxy -4 '-O- methyl capillarisins, structural formula are：
Artemetine, structure be for：
The preparation method and its usage of alpine yallow herb active component of the present invention, this method is stable, reliable, technique letter It is single, multiple monomer components can be obtained simultaneously, and purity is high, can prepare in batches.The compound of acquisition has anti-influenza virus activity, can It is that one kind can apply to prevent, treat flu, abscess of throat as the new resisiting influenza virus lead compound of high-efficiency low-toxicity Etc. the raw materials for production in the medicine, health products or food of disease.
The present invention does not only disclose the material composition of active component, it is often more important that actual solving is complicated in alpine yallow herb The problem of compound is difficult to obtain, and procurement cost is high, the monomer component obtained by the method for the invention is also to be further whole The resisiting influenza virus Effect study of body animal provides material guarantee, is the new resisiting influenza virus guide chemical combination of a kind of high-efficiency low-toxicity Thing.
Embodiment
The present invention is further described in conjunction with the embodiments
Embodiment 1
A. dry alpine yallow herb medicinal material 1kg is taken, is crushed, 10 mesh sieves is crossed, puts in round-bottomed flask, adds the volume of 8 times of weight Concentration is that 30% ethanol water is extracted, and 50 DEG C of water bath heating temperature, refluxing extraction 5 times, the time is 1 hour/time, extraction Filtrate afterwards is 1.2 through being concentrated under reduced pressure into relative density, and it is standby to obtain medicinal extract；
B. the medicinal extract that will be obtained in step a, vacuum dried case are dried, and vacuum -0.08MPa, temperature 45 C, obtain one Branch parthenium extract, it is standby；
C. extract step b obtained is nylon-6 resin by weight 1 with polyamide:6 are adsorbed, resin Post blade diameter length ratio is 1:3, sample solution extract concentrations are 4%, and loading flow velocity is 0.5BV/h, and polyamide is first eluted with water after absorption Resin 10BV, elution flow rate 0.5BV/h, then after water elution is discarded, the ethanol water ladder for being 10-95% with volumetric concentration Degree elution 5BV, flow velocity 0.5BV/h, the ethanol eluate that concentration is 30% gradient, rotated evaporimeter concentration are collected, then passed through Vacuum drying chamber dry, pulverize, and produce alpine yallow herb purified；
D. the alpine yallow herb purified that step c is obtained is suspended with water, successively with isometric petroleum ether:Chloroform extracts Take, collect chloroform extraction liquid, it is 1.0 that rotated evaporimeter, which is concentrated into relative density, and it is standby to obtain medicinal extract；
E. alpine yallow herb purified medicinal extract step d obtained is with silica gel with weight than 1:1 is adsorbed and is volatilized, silicagel column Blade diameter length ratio is 1:5, wet method is splined on silicagel column, successively with volume ratio 10:0-0:10 petroleum ether:Ethyl acetate gradient, Elution flow rate 0.5-3BV/h, thin-layer chromatography monitoring collection liquid, allied substances is merged, rotated evaporimeter is concentrated into phase respectively It is 1.5 to density, its pressure is -0.08MPa, and temperature is 40 DEG C, and solvent normal temperature volatilizes, and adding methanol makes dissolving, normal temperature volatilization To crystal is separated out, filtration, 3 monomeric compounds, respectively -4 '-O- of casticin 321mg, 6- de-methoxies methyl mattresses are obtained Old chromone 357mg and artemetine 63mg, content are respectively casticin 96.2%, 6- de-methoxy -4 '-O- methyl oriental wormwoods Chromone 97.5%, artemetine 95.4%.
Embodiment 2
A. dry alpine yallow herb medicinal material 1kg is taken, is crushed, 10 mesh sieves is crossed, puts in round-bottomed flask, adds the volume of 10 times of weight Concentration is that 50% ethanol water is extracted, and 60 DEG C of water bath heating temperature, refluxing extraction 3 times, the time is 2 hours/time, extraction Filtrate afterwards is 1.3 through being concentrated under reduced pressure into relative density, and it is standby to obtain medicinal extract；
B. the medicinal extract that will be obtained in step a, vacuum dried case are dried, and vacuum -0.08MPa, temperature 50 C, obtain one Branch parthenium extract, it is standby；
C. extract step b obtained is nylon -66 resin by weight 1 with polyamide:8 are adsorbed, tree Fat post blade diameter length ratio is 1:5, sample solution extract concentrations are 6%, and loading flow velocity is 1.0BV/h, and polyamides is first eluted with water after absorption Polyimide resin 8BV, elution flow rate 1.0BV/h, then after water elution is discarded, the ethanol for being 10-95% with volume by volume concentration is water-soluble Liquid gradient elution 8BV, flow velocity 1.0BV/h, the ethanol eluate that concentration is 50% gradient is collected, rotated evaporimeter concentrates, Vacuum dried case dry, pulverize again, produce alpine yallow herb purified；
D. the alpine yallow herb purified that step c is obtained is suspended with water, successively with petroleum ether, chloroform equal-volume extraction, Chloroform extraction liquid is collected, it is 1.1 that rotated evaporimeter, which is concentrated into relative density, and it is standby to obtain medicinal extract；
E. alpine yallow herb purified medicinal extract step d obtained is with silica gel with weight than 1:2 are adsorbed and are volatilized, silicagel column Blade diameter length ratio is 1:8, wet method is splined on silicagel column, successively with volume ratio 10:0-0:10 petroleum ether:Ethyl acetate gradient, Elution flow rate 0.5-3BV/h, thin-layer chromatography monitoring collection liquid, allied substances is merged, rotated evaporimeter is concentrated into phase respectively It is 1.3 to density, its pressure is -0.08MPa, and temperature is 50 DEG C, and solvent normal temperature volatilizes, and adding methanol makes dissolving, normal temperature volatilization To crystal is separated out, filtration, 3 monomeric compounds, respectively -4 '-O- of casticin 311mg, 6- de-methoxies methyl mattresses are obtained Old chromone 363mg and artemetine 69mg, content are respectively casticin 97.8%, 6- de-methoxy -4 '-O- methyl oriental wormwoods Chromone 98.9%, artemetine 96.7%.
Embodiment 3
A. dry alpine yallow herb medicinal material 1kg is taken, is crushed, 10 mesh sieves is crossed, puts in round-bottomed flask, adds the volume of 15 times of weight Concentration is that 75% ethanol water is extracted, and 80 DEG C of water bath heating temperature, refluxing extraction 2 times, the time is 1 hour/time, extraction Filtrate afterwards is 1.3 through being concentrated under reduced pressure into relative density, and it is standby to obtain medicinal extract；
B. the medicinal extract that will be obtained in step a, vacuum dried case are dried, and vacuum -0.08MPa, 65 DEG C of temperature, obtain one Branch parthenium extract, it is standby；
C. extract step b obtained is nylon-610 resin by weight 1 with polyamide:10 are adsorbed, Resin column blade diameter length ratio is 1:7, sample solution extract concentrations are 10%, and loading flow velocity is 3BV/h, and polyamides is first eluted with water after absorption Polyimide resin 5BV, elution flow rate 3BV/h, then after water elution is discarded, the ethanol gradient for being 10-95% with volume by volume concentration is washed De- 10BV, flow velocity 3BV/h, the ethanol eluate that concentration is 60% gradient, rotated evaporimeter concentration are collected, then done through vacuum Dry case dry, pulverize, and produce alpine yallow herb purified；
D. the alpine yallow herb purified that step c is obtained is suspended with water, successively with petroleum ether, chloroform equal-volume extraction, Chloroform extraction liquid is collected, it is 1.1 that rotated evaporimeter, which is concentrated into relative density, and it is standby to obtain medicinal extract；
E. alpine yallow herb purified medicinal extract step d obtained is with silica gel with 1:3 weight are adsorbed and volatilized, silicagel column than implementing Blade diameter length ratio is 1:10, wet method is splined on silicagel column, successively with volume ratio 10:0-0:10 petroleum ether:Ethyl acetate gradient, Elution flow rate 0.5-3BV/h, thin-layer chromatography monitoring collection liquid, allied substances is merged, rotated evaporimeter is concentrated into phase respectively It is 1.4 to density, its pressure is -0.08MPa, and temperature is 60 DEG C, and solvent normal temperature volatilizes, and adding methanol makes dissolving, normal temperature volatilization To crystal is separated out, filtration, 3 monomeric compounds, respectively -4 '-O- of casticin 355mg, 6- de-methoxies methyl mattresses are obtained Old chromone 378mg and artemetine 62mg, content are respectively casticin 97.1%, 6- de-methoxy -4 '-O- methyl oriental wormwoods Chromone 98.2%, artemetine 96.3%.
Embodiment 4
The method of the invention obtains the pharmacodynamic study of alpine yallow herb active component：
Using MDCK (dog kidney) cell as virus host, determination sample suppresses virus and causes cytopathy degree for this experiment (CPE)；
Material and method：
Strain：Influenza virus A/Chinese prevents/359/95 (H3N2), in chick embryo allantois intracavitary subculture (2013.9), -80 DEG C preserve；
Sample treatment：Sample is made into suitable initial concentration with dilution, then makees 3 times of dilutions, each 8 dilutions with nutrient solution Degree；
Positive control drug：Virazole (RBV), Xinxiang Pharmacy stock Co., Ltd (lot number 20081227), Tamiflu, China's food Product drug assay research institute (lot number 101096-200901)；
Method of testing：Mdck cell is inoculated with 96 well culture plates, puts 5%CO2, 37 DEG C of cultures.24 hours postoperative infection influenza diseases Malicious 1/310-5, adsorb 2 hours, abandon virus liquid, add the maintaining liquid containing different dilution factor samples and positive control drug, simultaneously If cell control well and virus control wells, 5%CO2, 37 DEG C of cultures of temperature are observed each when virus control group lesion (CPE) reaches 4+ Group cytopathy (CPE) (about 40 hours), calculate each sample resisiting influenza virus half-inhibition concentration (IC50).
Experimental result：
It the results are shown in Table 1；
The anti-influenza virus activity the selection result of table 1
Note：(1) "-" represents sample in maximal non-toxic dosage without anti influenza Alphavirus activity in table.(2)TC50：Medicine half The poisonous concentration of number；IC50：Medicine is to viral half-inhibition concentration；SI：Select index, SI=TC50/IC50。
Experiment conclusion：
Under the experimental condition, casticin, 6- de-methoxies -4 '-O- methyl capillarisin and artemetine are to influenza The viral A/ Chinese prevents that/359/95 (H3N2) has inhibitory activity.
Embodiment 5
The Structural Identification of the alpine yallow herb active component obtained by the method for the invention：
The identification of compound casticin：Casticin is Yellow amorphous powder, EI-MS m/z：374, with dividing Minor C19H18O8It is consistent；1H-NMR (DMSO, 500MHz) δ：7.67 (1H, d, J=2Hz, H-2 '), 7.62 (1H, dd, J=8.5, 1.5Hz, H-6 '), 6.96 (1H, d, J-6.8Hz, H-5 '), 6.91 (1H, s, H-8), 3.72~3.91 (12H, br, 4- OCH3).With reference to casticin standard items HPLC comparison results and document, it is casticin to determine the compound.Its structure Formula 1：
The identification of compound 6- de-methoxy -4 '-O- methyl capillarisins：White amorphous powder, EI-MS m/z： 300.With molecular formula C16H12O6It is consistent.1H-NMR (DMSO, 500MHz) δ：7.32 (1H, d, J=2.5Hz, H-2 '), 7.31 (1H, D, J=2Hz, H-6 '), 7.06 (1H, d, J=2Hz, H-5 '), 7.05 (1H, d, J=2Hz, H-3 '), 6.34 (1H, d, J= 2.5Hz, H-6), 6.19 (1H, d, J=2Hz, H-8), 5.04 (1H, s, H-3), 3.78 (1H, s ,-OCH3).Methoxy is removed with reference to 6- Base -4 '-O- methyl capillarisin standard items HPLC comparison results and document, it is -4 '-O- of 6- de-methoxies to determine the compound Methyl capillarisin.Its structure formula 2：
The identification of compound artemetine：Yellow needles, EI-MS m/z：388.With molecular formula C20H20O8It is consistent.1H- NMR (DMSO, 500MHz) δ：7.75 (1H, dd, J=6.5,2Hz, H-6 '), 7.69 (1H, d, J=2Hz, H-2 '), 7.08 (1H, D, J=6.8Hz, H-5 '), 6.77 (1H, s, H-8), 3.78~3.92 (15H, br, 5-an OCH3).With reference to artemetine standard items HPLC comparison results and document, it is artemetine to determine compound.Its structure formula 3：
Through Structural Identification：Respectively casticin content is between 96%-98%, 6- de-methoxy -4 '-O- methyl mattresses Old chromone content is between 97%-99%, and artemetine content is between 95%-97%.

Claims (3)

1. a kind of preparation method of alpine yallow herb active component, it is characterised in that follow these steps to carry out：
A, dry alpine yallow herb medicinal material is taken, through crushing, 10 mesh sieves is crossed, is placed in round-bottomed flask, add the volume of 8-15 times of weight Concentration is that 30-75% ethanol waters are extracted, 50-80 DEG C of water bath heating temperature, refluxing extraction 2-5 times, and the time is that 1-3 is small When/time, filtrate after extraction is 1.0-1.5 through being concentrated under reduced pressure into relative density, and it is standby to obtain medicinal extract；
B, the medicinal extract that will be obtained in step a, vacuum dried case are dried, and vacuum -0.08MPa, 45-70 DEG C of temperature, obtain one Branch parthenium extract, it is standby；
C, the extract for obtaining step b and polyamide be nylon-6 or nylon -66 or nylon-610 resin by weight 1:6-1:10 are adsorbed, and resin column blade diameter length ratio is 1:3-1:7, sample solution extract concentrations are 4%-10%, and loading flow velocity is 0.5-3 BV/h, polyamide 5-10BV, elution flow rate 0.5-3 BV/h are first eluted with water after absorption, then water elution is abandoned After going, the ethanol water for being 10-95% with volume by volume concentration elutes 5-10BV, and flow velocity is 0.5-3 BV/h, collects concentration and is The ethanol eluate of 30-60% gradients, rotated evaporimeter concentration, then vacuum dried case dry, pulverize, and it is pure to produce alpine yallow herb Compound；
D, the alpine yallow herb purified that step c is obtained is suspended with water, petroleum ether layer, three chloromethanes of water layer is discarded with petroleum ether extraction Alkane equal-volume extraction, chloroform extraction liquid is collected, it is 1.0-1.2 that rotated evaporimeter, which is concentrated into relative density, obtains medicinal extract It is standby；
E, the alpine yallow herb purified medicinal extract for obtaining step d and silica gel with weight than 1:1-1:3 are adsorbed and are volatilized, silicagel column Blade diameter length ratio is 1:5-1:10, wet method is splined on silicagel column, successively with volume ratio 10:0-0:10 petroleum ether:Ethyl acetate gradient Elution, elution flow rate 0.5-3 BV/h, thin-layer chromatography monitoring collection liquid, allied substances are merged, rotated evaporimeter is dense respectively It is 1.0-1.5 to be reduced to relative density, and pressure is -0.08MPa, and temperature is 40 DEG C -60 DEG C, and solvent normal temperature volatilizes, and it is molten to add methanol Solution, normal temperature evaporates into precipitation crystal, filters, obtains monomeric compound, is respectively casticin through Structural Identification, yield 0.30mg/g, content is between 96%-98%；6- de-methoxy -4 '-O- methyl capillarisins, yield 0.35mg/g, content exist Between 97%-99%；Artemetine, yield 0.06mg/g, content is between 95%-97%.
2. the alpine yallow herb active component that method according to claim 1 obtains prepare the prevention resisiting influenza virus A/ Chinese it is anti-/ Purposes in 359/95 (H3N2) medicine.
3. the alpine yallow herb active component that method according to claim 1 obtains prepare the resisiting influenza virus A/ Chinese it is anti-/ Purposes in 359/95 (H3N2) health products.
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