CN103664862B - Polyphenolic compound and its preparation method and application in a kind of Turkish tobaccos - Google Patents

Polyphenolic compound and its preparation method and application in a kind of Turkish tobaccos Download PDF

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CN103664862B
CN103664862B CN201310378439.3A CN201310378439A CN103664862B CN 103664862 B CN103664862 B CN 103664862B CN 201310378439 A CN201310378439 A CN 201310378439A CN 103664862 B CN103664862 B CN 103664862B
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polyphenolic compound
silica gel
medicinal extract
compound
rhizome
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CN103664862A (en
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陈章玉
韩熠
张霞
张涛
杨光宇
刘志华
陈永宽
缪明明
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Yunnan Academy of Tobacco Science
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Yunnan Academy of Tobacco Science
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D313/00—Heterocyclic compounds containing rings of more than six members having one oxygen atom as the only ring hetero atom
    • C07D313/02—Seven-membered rings
    • C07D313/06—Seven-membered rings condensed with carbocyclic rings or ring systems
    • C07D313/10—Seven-membered rings condensed with carbocyclic rings or ring systems condensed with two six-membered rings
    • C07D313/14—[b,f]-condensed
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
    • A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
    • A01N43/22—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom rings with more than six members

Abstract

The invention discloses polyphenolic compound and its preparation method and application in a kind of Turkish tobaccos.Described polyphenolic compound is separated to obtain from Turkish tobaccos rhizome, and its molecular formula is C 18h 18o 6, there is following structure: the preparation method of described polyphenolic compound is with Turkish tobaccos rhizome for raw material, through medicinal extract extraction, silica gel column chromatography, high pressure liquid chromatography separating step, be specially: with ethanol ultrasonic extraction after Turkish tobaccos rhizome is pulverized, extracting solution merges, and filter, concentrating under reduced pressure becomes medicinal extract; Medicinal extract silica gel dry column-packing carries out silica gel column chromatography; Be that the chloroform-acetone solution of 1:0 ~ 1:2 carries out gradient elution with volume proportion; Namely the 7:3 part of elutriant obtains described polyphenolic compound with high pressure liquid chromatography separation and purification further.The present invention proves to have good cytoactive to tobacco mosaic virus (TMV) through test.The compounds of this invention structure is simple, and synthetic easily realizes, and the activity of compound is good, can be used as the guiding compound of resisting tobacco mosaic virus.

Description

Polyphenolic compound and its preparation method and application in a kind of Turkish tobaccos
Technical field
The invention belongs to tobacco chemistry field, be specifically related to a kind of extract from Turkish tobaccos rhizome polyphenolic compound, its preparation method and application.
Background technology
Tobacco is one the abundantest containing chemical substance in each kind of plant of being familiar with of the mankind, and through the research of decades, the monomer chemistries material that people identify out at present from tobacco just more than kind more than 3000, and also has many compositions not yet to identify out.Tobacco, except being mainly used in cigarette smoking purposes, also therefrom can extract the multiple chemical composition having utility value, therefrom finds that there is the guiding compound of value of exploiting and utilizing.Particularly for the root of tobacco and stem, general all as the offal treatment in leaf tobacco production process.Therefore, except as except cigarette consumption, the comprehensive utilization strengthening tobacco and waste thereof is significant.
Tobacco aldehydes matter (comprising phenol and derivative thereof) is formed by the metabolism in cigarette strain body between three large materials (carbohydrate, lipid, protein), is one of important secondary substance of tobacco.Polyphenolic substance not only has material impact to tobacco leaf color and luster, fragrance and flue gas physiological strength, or weighs the important indicator of cigarette quality; And polyphenolic compound also has pharmacological action widely, as: antitumor, anti-human immunodeficiency virus (HIV), anti-oxidant, antibacterial, anticoagulation etc.; Existing research simultaneously confirms, its pharmacological action and chemical structure closely related, more polyphenolic compound can be researched and developed further, therefrom find effective lead compound and active group.The present invention is separated and obtains a kind of new polyphenolic compound with activity of resisting tobacco mosaic virus from Turkish tobaccos rhizome, and this compound it is not yet seen relevant report.
Summary of the invention
The first object of the present invention is to provide the polyphenolic compound in a kind of Turkish tobaccos rhizome; Second object is the preparation method providing this polyphenolic compound; 3rd object is that described polyphenolic compound is preparing the application in resisting tobacco mosaic disease medicine.
The first object of the present invention is achieved in that described compound is separated to obtain from Turkish tobaccos rhizome, and its molecular formula is C 18h 18o 6, there is following structure:
。
This compound is yellow jelly, and chemical name is Turkish tobaccos phenol A(Turktobpheonl A).
The second object of the present invention is achieved in that the preparation method of described polyphenolic compound is with Turkish tobaccos rhizome for raw material, through medicinal extract extraction, silica gel column chromatography, high pressure liquid chromatography separating step, is specially:
A, medicinal extract extract: get Turkish tobaccos rhizome, be crushed to 20 ~ 40 orders, with 80-100% ethanol ultrasonic extraction 3 ~ 5 times, and each 30 ~ 60min, united extraction liquid, filtration, concentrating under reduced pressure becomes medicinal extract;
B, silica gel column chromatography: 100 ~ 200 order silica gel dry column-packings of medicinal extract weight ratio 2.5 times amount carry out silica gel column chromatography; Be that the chloroform-acetone solution of 1:0 ~ 1:2 carries out gradient elution with volume proportion, merge identical part, collect each several part elutriant and concentrated;
C, high pressure liquid chromatography are separated: namely the 7:3 part of step B elutriant obtains described polyphenolic compound with high pressure liquid chromatography separation and purification further.
The structure of the polyphenolic compound prepared with aforesaid method identifies out by the following method:
The compounds of this invention is yellow jelly; UV spectrum (solvent is methyl alcohol), λ max(log ε) 315(3.92), 236(4.12), 210(4.32); Infrared spectra (pressing potassium bromide troche) ν max3412,3028,2965,2875,1612,1586,1524,1458,1429,1398,1185,1153,1126,1078,855cm -1; High resolution mass spectrum (HRESIMS) provides quasi-molecular ion peak m/z[353.1008 M+Na] +(calculated value 353.1001).In conjunction with 1h and 13c NMR spectrum provides a molecular formula C 18h 18naO 6, degree of unsaturation is 10.From 1h and 13cNMR composes (attribution data is in Table-1) signal can find out in compound have 1,2,5, a 6-tetra-substituted benzene [C-1 ~ C-6; δ c145.1,141.0,115.5,106.5,150.5 and 120.9; δ h6.89 d( j=8.8) and 6.60 d( j=8.8)]; 1', 2', 3', 4', a 6'-five substituted benzene (C-1' ~ C-6'; δ c136.4,154.0,112.9,149.4,104.1,127.8; δ h6.52s); One group of double bond [CH-7 and CH-8, δ c126.0 and 130.0; δ h7.07d( j=11.6) and 6.79d( j=11.6)]; An ethanol based [CH 2-7' and CH 2-8'; δ c35.6,63.6; δ h2.59 t(7.2), 3.64 t(7.2)]; Two methoxyl groups ( δ c55.9,61.3; δ h3.80s, 3.87s); Two phenolic hydroxyl groups ( δ c9.43brs, 9.62brs).Carefully analyzing this compound of these data acknowledgements is dibenz [b, f] oxepin compounds.From HMBC spectrum (accompanying drawing-3); H-7'( δ h2.59) and C-2'( δ c154.0), C-3'( δ c112.9), C-4'( δ c149.4), H-8'( δ h3.64) and C-3'( δ c112.9) existence is significantly correlated with; This confirms that ethanol based is substituted in C-3'.Two methoxyl group proton signals ( δ h3.80,3.87) respectively and C-5( δ c150.5), C-2 ' ( δ c154.0) two methoxy substitutions are in C-5 and C-2 ' position to have obvious coherent signal to confirm.Phenolic hydroxyl group proton signal ( δ h9.43) and C-1( δ c145.1), C-2( δ c141.0), C-3( δ c115.5) this phenolic hydroxyl group is substituted in C-2 position to have coherent signal to confirm; Another phenolic hydroxyl group signal ( δ h9.26) and C-3'( δ c112.9), C-4'( δ c149.4), C-5'( δ c104.1) relevant another phenolic hydroxyl group signal of confirmation is further had to be substituted in C-4'.So far the structure of this compound is determined.
Table-1. compounds 1h NMR and 13c NMR data (C 5d 5n)
No. δ C（m） δ H（m, J, Hz）
1 145.1 s
2 141.0 s
3 115.5 d 6.89 d（8.8）
4 106.5 d 6.60 d（8.8）
5 150.5 s
6 120.9 s
7 126.0 d 7.07 d（11.6）
8 130.0 d 6.79 d（11.6）
1＇ 136.4 s
2＇ 154.0 s
3＇ 112.9 d
4＇ 149.4 s
5＇ 104.1 d 6.52 s
6＇ 127.8 s
7＇ 35.6 t 2.59 t（7.2）
8＇ 63.6 t 3.64 t（7.2）
5-OMe 55.9 q 3.80 s
2＇-OMe
4＇-OMe 61.3 q 3.87 s
2-Ar-OH 9.43 brs
2＇-Ar-OH 9.62 brs
4＇-Ar-OH
The third object of the present invention is achieved in that the preparation be applied to by described polyphenolic compound in Tobacco mosaic medicine.
Compounds of the present invention is separated first, is determined as polyphenolic compound by nucleus magnetic resonance and measuring method of mass spectrum, and characterizes its concrete structure.Through the experiment to resisting tobacco mosaic virus, its relative inhibition reaches 45.6%, has good activity of resisting tobacco mosaic virus, higher than the relative inhibition (30.8%) of positive reference substance Nanning mycin.Above result discloses compound of the present invention preparing in resisting tobacco mosaic virus medicine good application prospect.The compounds of this invention structure is simple active good, can be used as the guiding compound of resisting tobacco mosaic virus medicine.
Accompanying drawing explanation
Fig. 1 is the carbon-13 nmr spectra of compound;
Fig. 2 is the proton nmr spectra of compound;
Fig. 3 is that the main HMBC of compound is correlated with.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is further illustrated, but limited the present invention never in any form, and any conversion done based on training centre of the present invention or improvement, all fall into protection scope of the present invention.
Except as otherwise noted, the percentage ratio adopted in the present invention is mass percent.
The preparation method C of polyphenolic compound of the present invention 18h 18o 6preparation method comprise medicinal extract extraction, silica gel column chromatography, high pressure liquid chromatography separating step, specifically comprise:
A, medicinal extract extract: get Turkish tobaccos rhizome, be crushed to 20 ~ 40 orders, with 80-100% ethanol ultrasonic extraction 3 ~ 5 times, and each 30 ~ 60min, united extraction liquid, filtration, concentrating under reduced pressure becomes medicinal extract;
B, silica gel column chromatography: 160 ~ 200 order silica gel dry column-packings of medicinal extract weight ratio 2.5 times amount carry out silica gel column chromatography; Be that the chloroform-acetone solution of 1:0 ~ 1:2 carries out gradient elution with volume proportion, merge identical part, collect each several part elutriant and concentrated;
C, high pressure liquid chromatography are separated: namely the 7:3 part of step B elutriant obtains described polyphenolic compound with high pressure liquid chromatography separation and purification further.
The etoh solvent concentration of described step A is 95%.
The medicinal extract of described step B, before silica gel column chromatography rough segmentation, uses weight ratio 0.8 ~ 1.2 times of 80 ~ 100 order silica gel mixed sample with after the pure dissolve with methanol of weight ratio 1.5 ~ 3 times amount.
The chloroform-acetone solution volume proportion of described step B is 1:0,20:1,9:1,8:2,7:3,6:4,1:1,1:2.
High performance liquid chromatography separation and purification in described step C adopts 21.2mm × 250mm, the C of 5 μm 18chromatographic column, flow velocity is 20mL/min, and moving phase is the methyl alcohol of 48%, and UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collects the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness.
Material after the separation and purification of described step C mesohigh liquid phase chromatography uses pure dissolve with methanol again, then with pure methyl alcohol for moving phase, is separated with gel filtration chromatography, with further separation and purification.
Polyphenolic compound of the present invention is preparing the application in resisting tobacco mosaic virus medicine.
The present invention is raw materials used not to be limited by area and kind, and all can realize the present invention, to derive from Baoshan Turkish tobaccos rhizome sample, the present invention will be further described below:
Embodiment 1
Turkish tobaccos rhizome sample source is in Baoshan, Yunnan, and kind is Bath horse.Turkish tobaccos rhizome is sampled 1.8 kg and is crushed to 30 orders, the supersound extraction 3 times of the ethanol with 90%, extracts 35min at every turn, and extracting solution merges, and filter, concentrating under reduced pressure becomes medicinal extract, obtains medicinal extract 86.5g.With the thick silica gel mixed sample of 100 order of 150g after the pure dissolve with methanol of medicinal extract weight ratio 2.5 times amount, the 160 order silica gel dress posts of 1.5kg carry out silica gel column chromatography, be 1:0 with volume proportion, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, chloroform-acetone the gradient elution of 1:2, TLC monitoring merges identical part, obtain 8 parts, wherein volume proportion is the prompt logical sequence 1,100 half preparative high-performance liquid chromatographic separation of chloroform-acetone elution fraction peace of 7:3, methyl alcohol with 48% is moving phase, Zorbax SB-C18(21.2 × 250mm, 5 μm) preparative column is stationary phase, flow velocity is 20ml/min, UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collect the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness, products therefrom uses pure dissolve with methanol again, then with pure methyl alcohol for moving phase, is separated, obtains this new compound with Sephadex LH-20 gel filtration chromatography.
Embodiment 2
Turkish tobaccos rhizome sample source is in Baoshan, Yunnan, and kind is Bath horse.Turkish tobaccos rhizome is sampled 1.9 kg and is crushed to 20 orders, the supersound extraction 5 times of the ethanol with 99%, extracts 60min at every turn, and extracting solution merges, and filter, concentrating under reduced pressure becomes medicinal extract, obtains medicinal extract 86.5g.With the thick silica gel mixed sample of 80 order of 150g after the pure dissolve with methanol of medicinal extract weight ratio 1.5 times amount, the 100 order silica gel dress posts of 1.5kg carry out silica gel column chromatography, be 1:0 with volume proportion, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, chloroform-acetone the gradient elution of 1:2, TLC monitoring merges identical part, obtain 8 parts, wherein volume proportion is the prompt logical sequence 1,100 half preparative high-performance liquid chromatographic separation of chloroform-acetone elution fraction peace of 7:3, methyl alcohol with 48% is moving phase, Zorbax SB-C18(21.2 × 250mm, 5 μm) preparative column is stationary phase, flow velocity is 20ml/min, UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collect the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness, products therefrom uses pure dissolve with methanol again, then with pure methyl alcohol for moving phase, is separated, obtains this new compound with Sephadex LH-20 gel filtration chromatography.
Embodiment 3
Turkish tobaccos rhizome sample source is in Baoshan, Yunnan, and kind is Samsun.Turkish tobaccos rhizome sampling 2.2kg is crushed to 40 orders, and the supersound extraction 3 times of the ethanol with 90%, extracts 30min at every turn, and extracting solution merges, and filter, concentrating under reduced pressure becomes medicinal extract, obtains medicinal extract 102.4g.With the thick silica gel mixed sample of 100 order of 180g after the pure dissolve with methanol of medicinal extract weight ratio 2.0 times amount, the 160 order silica gel dress posts of 1.8 kg carry out silica gel column chromatography, be 1:0 with volume proportion, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, chloroform-acetone the gradient elution of 1:2, TLC monitoring merges identical part, obtain 8 parts, wherein volume proportion is the prompt logical sequence 1,100 half preparative high-performance liquid chromatographic separation of chloroform-acetone elution fraction peace of 7:3, methyl alcohol with 48% is moving phase, Zorbax SB-C18(21.2 × 250mm, 5 μm) preparative column is stationary phase, flow velocity is 20ml/min, UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collect the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness, products therefrom uses pure dissolve with methanol again, then with pure methyl alcohol for moving phase, is separated, obtains this new compound with Sephadex LH-20 gel filtration chromatography.
Embodiment 4
Turkish tobaccos rhizome sample source is in Baoshan, Yunnan, and kind is Samsun.Turkish tobaccos rhizome sampling 2.1kg is crushed to 40 orders, and the supersound extraction 3 times of the ethanol with 94%, extracts 40min at every turn, and extracting solution merges, and filter, concentrating under reduced pressure becomes medicinal extract, obtains medicinal extract 102.4g.With the thick silica gel mixed sample of 100 order of 180g after the pure dissolve with methanol of medicinal extract weight ratio 3 times amount, the 200 order silica gel dress posts of 1.8kg carry out silica gel column chromatography, be 1:0 with volume proportion, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, chloroform-acetone the gradient elution of 1:2, TLC monitoring merges identical part, obtain 8 parts, wherein volume proportion is the prompt logical sequence 1,100 half preparative high-performance liquid chromatographic separation of chloroform-acetone elution fraction peace of 7:3, methyl alcohol with 48% is moving phase, Zorbax SB-C18(21.2 × 250mm, 5 μm) preparative column is stationary phase, flow velocity is 20ml/min, UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collect the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness, products therefrom uses pure dissolve with methanol again, then with pure methyl alcohol for moving phase, is separated, obtains this new compound with Sephadex LH-20 gel filtration chromatography.
Embodiment 5
Turkish tobaccos rhizome sample source is in Baoshan, Yunnan, and kind is section Kuru.Turkish tobaccos rhizome sampling 2.0kg is crushed to 35 orders, and the supersound extraction 3 times of the ethanol with 95%, extracts 40min at every turn, and extracting solution merges, and filter, concentrating under reduced pressure becomes medicinal extract, obtains medicinal extract 96.4g.With the thick silica gel mixed sample of 100 order of 160g after the pure dissolve with methanol of medicinal extract weight ratio 2.2 times amount, the 160 order silica gel dress posts of 1.6kg carry out silica gel column chromatography, be 1:0 with volume proportion, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, chloroform-acetone the gradient elution of 1:2, TLC monitoring merges identical part, obtain 8 parts, wherein volume proportion is the prompt logical sequence 1,100 half preparative high-performance liquid chromatographic separation of chloroform-acetone elution fraction peace of 7:3, methyl alcohol with 48% is moving phase, Zorbax SB-C18(21.2 × 250mm, 5 μm) preparative column is stationary phase, flow velocity is 20ml/min, UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collect the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness, products therefrom uses pure dissolve with methanol again, then with pure methyl alcohol for moving phase, is separated, obtains this new compound with Sephadex LH-20 gel filtration chromatography.
Embodiment 6
Turkish tobaccos rhizome sample source is in Baoshan, Yunnan, and kind is section Kuru.Turkish tobaccos rhizome sampling 2.5kg is crushed to 30 orders, and the supersound extraction 4 times of the ethanol with 96%, extracts 40min at every turn, and extracting solution merges, and filter, concentrating under reduced pressure becomes medicinal extract, obtains medicinal extract 96.4g.With the thick silica gel mixed sample of 100 order of 160g after the pure dissolve with methanol of medicinal extract weight ratio 2.5 times amount, the 160 order silica gel dress posts of 1.6kg carry out silica gel column chromatography, be 1:0 with volume proportion, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1, chloroform-acetone the gradient elution of 1:2, TLC monitoring merges identical part, obtain 8 parts, wherein volume proportion is the prompt logical sequence 1,100 half preparative high-performance liquid chromatographic separation of chloroform-acetone elution fraction peace of 7:3, methyl alcohol with 48% is moving phase, Zorbax SB-C18(21.2 × 250mm, 5 μm) preparative column is stationary phase, flow velocity is 20ml/min, UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collect the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness, products therefrom uses pure dissolve with methanol again, then with pure methyl alcohol for moving phase, is separated, obtains this new compound with Sephadex LH-20 gel filtration chromatography.
Embodiment 7
Compound prepared by Example 1 is yellow jelly.
Measuring method is: with nucleus magnetic resonance, in conjunction with other spectroscopic technique qualification structure.
1) UV spectrum (solvent is methyl alcohol), λ max(log ε) 315(3.92), 236(4.12), 210(4.32) nm;
2) infrared spectra (pressing potassium bromide troche) ν max3412,3028,2965,2875,1612,1586,1524,1458,1429,1398,1185,1153,1126,1078,855cm -1;
3) high resolution mass spectrum (HRESIMS) provides quasi-molecular ion peak m/z[353.1008 M+Na] +(calculated value 353.1001).In conjunction with 1h and 13c NMR spectrum provides a molecular formula C 18h 18naO 6, degree of unsaturation is 10.
4) from 1h and 13cNMR composes (attribution data is in Table-1) signal can find out in compound have 1,2,5, a 6-tetra-substituted benzene [C-1 ~ C-6; δ c145.1,141.0,115.5,106.5,150.5 and 120.9; δ h6.89 d( j=8.8) and 6.60 d( j=8.8)]; 1', 2', 3', 4', a 6'-five substituted benzene (C-1' ~ C-6'; δ c136.4,154.0,112.9,149.4,104.1,127.8; δ h6.52 s); One group of double bond [CH-7 and CH-8, δ c126.0 and 130.0; δ h7.07 d( j=11.6) and 6.79 d( j=11.6)]; An ethanol based [CH 2-7' and CH 2-8'; δ c35.6,63.6; δ h2.59 t(7.2), 3.64 t(7.2)]; Two methoxyl groups ( δ c55.9,61.3; δ h3.80s, 3.87s); Two phenolic hydroxyl groups ( δ c9.43brs, 9.62brs).Carefully analyzing these data acknowledgement compounds is dibenz [b, f] oxepin compounds.From HMBC spectrum (accompanying drawing 3); H-7'( δ h2.59) and C-2'( δ c154.0), C-3'( δ c112.9), C-4'( δ c149.4), H-8'( δ h3.64) and C-3'( δ c112.9) existence is significantly correlated with; This confirms that ethanol based is substituted in C-3 position C-3'.Two methoxyl group proton signals ( δ h3.80,3.87) respectively and C-5( δ c150.5), C-2 ' ( δ c154.0) two methoxy substitutions are at C-5 and C-2 ' to have obvious coherent signal to confirm.Phenolic hydroxyl group proton signal ( δ h9.43) and C-1( δ c145.1), C-2( δ c141.0), C-3( δ c115.5) this phenolic hydroxyl group is substituted in C-2 position to have coherent signal to confirm; Another phenolic hydroxyl group signal ( δ h9.26) and C-3'( δ c112.9), C-4'( δ c149.4), C-5'( δ c104.1) relevant another phenolic hydroxyl group signal of confirmation is further had to be substituted in C-4'.So far the structure of this compound is determined.
Embodiment 8
Compound prepared by Example 2 ~ 6 is yellow jelly.Authentication method is identical with embodiment 7, confirms that compound prepared by embodiment 2 ~ 6 is described polyphenolic compound---Turkish tobaccos phenol A(Turktobpheonl A).
Embodiment 9
Arbitrary polyphenolic compound prepared by Example 1 ~ 6 carries out activity of resisting tobacco mosaic virus test, and test situation is as follows:
Adopt half leaf method, when the mass concentration of medicament is 50 mg/L, activity of resisting tobacco mosaic virus mensuration is carried out to the compounds of this invention.5 ~ 6 age flue-cured tobacco plant on, choose the blade (leaf capable normal, anosis without worm) being applicable to test, first blade evenly sprinkled fine emery powder, with writing brush by tobacco mosaic virus (TMV) source (3.0 × 10 for subsequent use -3) be evenly put on sprinkled with silicon carbide blade on, connect after poison terminates until the blade of all middle choosings, be placed on immediately in the culture dish filling liquid and process 20min, take out, to spill on defoliation sheet the globule and about liquid, two and half leaves are restored and is emitted on the enamel son of an influential official cover glass being covered with toilet paper moisturizing, temperature control (23 ± 2) DEG C, be placed on greenhouse natural light irradiation, 2 ~ 3d and visible withered spot. each process sets second half leaf as contrast, be provided with in addition 1 group be the process of commodity Ningnanmycin as a comparison, by following formulae discovery relative inhibition.
XI%=（CK-T）／CK×100%
X: relative inhibition (%), CK: be soaked in the withered spot number (individual) that half in clear water connects malicious leaf, T is soaked in the withered spot number (individual) that half in liquid connects malicious leaf.
Result shows that the relative inhibition of this compound is 45.6%, far above the relative inhibition 30.8% of contrast Ningnanmycin, illustrates that compound has good activity of resisting tobacco mosaic virus.

Claims (7)

1. a polyphenolic compound, it is characterized in that described polyphenolic compound is separated to obtain from Turkish tobaccos rhizome, its molecular formula is C 18h 18o 6, there is following structure:
。
2. a preparation method for polyphenolic compound according to claim 1, is characterized in that with Turkish tobaccos rhizome for raw material, through medicinal extract extraction, silica gel column chromatography, high pressure liquid chromatography separating step, is specially:
A, medicinal extract extract: get Turkish tobaccos rhizome, be crushed to 20 ~ 40 orders, with 80 ~ 100% ethanol ultrasonic extraction 3 ~ 5 times, and each 30 ~ 60min, united extraction liquid, filtration, concentrating under reduced pressure becomes medicinal extract;
B, silica gel column chromatography: 100 ~ 200 order silica gel dry column-packings of medicinal extract weight ratio 2 ~ 3 times amount carry out silica gel column chromatography; Be that the chloroform-acetone solution of 1:0 ~ 1:2 carries out gradient elution with volume proportion, merge identical part, collect each several part elutriant and concentrated;
C, high pressure liquid chromatography are separated: namely the 7:3 part of step B elutriant obtains described polyphenolic compound with high pressure liquid chromatography separation and purification further.
3. the preparation method of polyphenolic compound as claimed in claim 2, is characterized in that in described step B, medicinal extract is before silica gel column chromatography rough segmentation, uses weight ratio 0.8 ~ 1.2 times of 80 ~ 100 order silica gel mixed sample with after the pure dissolve with methanol of weight ratio 1.5 ~ 3 times amount.
4. the preparation method of polyphenolic compound as claimed in claim 2, is characterized in that the chloroform-acetone solution volume proportion in described step B is 1:0,20:1,9:1,8:2,7:3,6:4,1:1,1:2.
5. the preparation method of polyphenolic compound as claimed in claim 2, is characterized in that in described step C, high performance liquid chromatography separation and purification adopts 21.2mm × 250mm, the C of 5 μm 18chromatographic column, flow velocity is 20mL/min, and moving phase is the methyl alcohol of 48%, and UV-detector determined wavelength is 254nm, each sample introduction 200 μ L, collects the chromatographic peak of 15.6min, repeatedly cumulative rear evaporate to dryness.
6. the preparation method of polyphenolic compound as claimed in claim 2, it is characterized in that the material after the separation and purification of described step C mesohigh liquid phase chromatography uses pure dissolve with methanol again, again with pure methyl alcohol for moving phase, with gel filtration chromatography be separated, with further separation and purification.
7. a polyphenolic compound according to claim 1 is preparing the application in resisting tobacco mosaic virus medicine.
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