CN106565724A - Dicranostigma leptopodum extract and extraction method and application thereof - Google Patents
Dicranostigma leptopodum extract and extraction method and application thereof Download PDFInfo
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- C07—ORGANIC CHEMISTRY
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- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/04—Ortho-condensed systems
- C07D491/056—Ortho-condensed systems with two or more oxygen atoms as ring hetero atoms in the oxygen-containing ring
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- 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
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
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- C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
- C07D491/14—Ortho-condensed systems
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Abstract
The invention relates to dicranostigma leptopodum extract and an extraction method and application thereof. Based on researches on the antimicrobial activity and active ingredients of dicranostigma leptopodum, it is found that the dicranostigma leptopodum extract has remarkable inhibitory activity for various plant pathogens and has the potential to prepare a plant antibacterial agent as an effective ingredient. Meanwhile, it is also found that the dicranostigma leptopodum extract contains multiple effective ingredients with magnoflorine as a majority. The invention further provides a method and process for quickly gathering the effective ingredients in the dicranostigma leptopodum.
Description
Technical field
The present invention relates to the medicine of the anti-phytopathogen of plant source, and in particular to a kind of favus of the scalp flower extract and its extraction side
Method and application.
Background technology
Phytopathogen has very serious shadow to the healthy growth and development, the yield of crops and quality of plant
Ring.Therefore, applying for bactericide has great importance in modern agriculture.But, the long-term weight of same bactericide
Multiple use already leads to phytopathogen and generates very strong drug resistance to many existing bactericides.Additionally, many countries
Some have been prohibitted the use of to environment, people or ecological with dysgenic disinfectant use in agriculture.Therefore, research and develop efficiently low
The Huang Dezhi of malicious and environmental protection has great importance.
Biological pesticide is that current novel pesticide studies most popular field, and botanical pesticide is one of biological pesticide important
Aspect.Slenderstalk dicranostigma herb（Dicranostigma leptopodum (maxim.）fedde）Also known as baldhead spends, reins in the horse back（Shaanxi）, rabbit
Son flower etc., belongs to Papaveraceae（Papaveraceae）One of convex combination algorithm plant.The plant be distributed widely in northwestern Yunnan Province, four
River western part, southern Tibet, East of Qinghai Province, SOUTH OF GANSU to the southeast, Qinling Mountains in Shaanxi north slope, Southern Shanxi Province, the Hebei west and south and
The Henan northwestward, is born in height above sea level 400-2900（-3700）The careless slope or roadside of rice, ridge, the top of a wall, roof are also common.Its root or
Herb can make medicinal, the effects such as have clearing heat and detoxicating, Repercusion analgesia, desinsection, control acute toothache, sore-throat, tonsillitis, lymph node
Core, the favus of the scalp, boil mange, ulcer.The plant is rich in various alkaloids, has been demonstrated with sedation-analgesia, antitumor, antibacterial etc.
Multiple biological activities.But, up to the present, there is not yet any grinding with regard to the anti-phytopathogen activity of favus of the scalp flower extract
Study carefully report.
Although slenderstalk dicranostigma herb has abundant natural resources in China, fail to be utilized effectively for a long time.This may be main
It is relevant the plant not to be eaten with humans and animals.Therefore, the present invention both provides one effectively for the exploitation of slenderstalk dicranostigma herb
Approach, also provide a kind of agricultural antiseptic in novel plant source for China, at the same also for vast farmerses provide one it is new
Get rich approach.
The content of the invention
It is an object of the invention to provide a kind of favus of the scalp flower extract and its extracting method and application, such medicine is to many plantations
Thing pathogen has good inhibitory activity.
The technical solution adopted in the present invention is：
Favus of the scalp flower extract, it is characterised in that：
The extract is Papaveraceae（Papaveraceae）Convex combination algorithm（Dicranostigma）Herbaceos perennial slenderstalk dicranostigma herb
（Dicranostigma leptopodum (maxim.）fedde）Herb or aerial part alcohol extract or water extract.
Include antimicrobial component, including magnoflorine, Chelerythrine, sanguinarine, corydalis in the favus of the scalp flower extract
Determine alkali, isocorydine, coptisine and Biflorine.
The application of favus of the scalp flower extract as mentioned, it is characterised in that：
The extract is used as the application for preparing plants antimicrobial medicine.
The extract has significant inhibitory activity to following phytopathogen：
Apple decay pathogen；Gibberella saubinetii pathogen；Early blight of tomato opportunistic pathogen；Curvularia pathogen；Pumpkin is withered cause of disease
Bacterium；Cotton is withered, withered germ of water-melon；Dry rot of potato opportunistic pathogen；Alternaria brassicae；Tobacco brown spot pathogen；Apple anthrax
Germ；Rice blast fungus；Botryosphaeria berengeriana f. sp；Tomato Cercospora Sojina Hara.
The extracting method of favus of the scalp flower extract as mentioned, it is characterised in that：
Comprise the following steps：
Step one：With ethanol, methyl alcohol or water as solvent, slenderstalk dicranostigma herb vegetable material is carried out heating extraction, merge what is repeatedly extracted
Extract, is stored at room temperature overnight, and decompression filters sediment, and gained filtrate decompression is evaporated off into solvent, obtains extract；
Step 2：Methanol aqueous solution is dissolved in by extract obtained, oil is used with petroleum ether, chloroform and n-butanol or successively successively
Ether, n-butanol are extracted；The n-butyl alcohol extract obtained by chloroform extract and n-butyl alcohol extract or rear method obtained by front method
As favus of the scalp flower extract.
In step：
Slenderstalk dicranostigma herb vegetable material is 1 g with the solid-liquid mixing ratio of solvent:15 mL, solution temperature is 50-80 DEG C, is stirred during dissolving
Or ultrasonic assistant；
Temperature when extract removes solvent under reduced pressure is 30～40 DEG C.
In step 2：
The extract of slenderstalk dicranostigma herb vegetable material is dissolved in methanol-water identical in quality with vegetable material, volume fraction is 10%～25%
In solution, then successively with petroleum ether, chloroform and n-butanol or petroleum ether, extracting n-butyl alcohol, every kind of solvent extraction three times；
During petroleum ether extraction, each solvent load is identical with the volume for being extracted liquid；
During chloroform extraction, each solvent load is the half for being extracted liquid product；
During extracting n-butyl alcohol, three solvent loads are followed successively by and are extracted 50%, 40% and the 30% of liquid product.
The present invention has advantages below：
Plants antimicrobial medicine using favus of the scalp flower extract as active ingredient involved in the present invention is a kind of botanical pesticide, category
In one kind of bio-pharmaceutical, meet requirement of the modern society to novel pesticide.Secondly, the medicine has that antibacterial activity is strong, antimicrobial spectrum
Extensively, environmental friendliness, be not likely to produce resistance, it is relatively low to humans and animals toxicity the features such as.Third, the plant favus of the scalp involved by the medicine
Flower has aboundresources, the characteristics of development cost is low.Fourth, the slenderstalk dicranostigma herb extraction and separation technology involved by the invention has production
Process is simple, low production cost, environmental pollution are little and are suitable for the advantage of large-scale production.
Description of the drawings
Fig. 1 is the high-efficient liquid phase chromatogram of slenderstalk dicranostigma herb alcohol extracts.
Specific embodiment
With reference to specific embodiment, the present invention will be described in detail.
The present invention relates to a class favus of the scalp flower extract and its medicine and application containing the extract or active ingredient, while
It is related to the extraction separation method of slenderstalk dicranostigma herb active ingredient.The present invention is based on to favus of the scalp flower extract antibacterial activity and active component
Research, it is found that favus of the scalp flower extract has significant inhibitory activity to various plants pathogen, with being used for as active ingredient
Prepare the potentiality of plant antimicrobial.Meanwhile, the invention is also found in favus of the scalp flower extract containing multiple based on magnoflorine
Active ingredient, and there is provided the method and technique of active ingredient in a kind of fast separating concentration slenderstalk dicranostigma herb.
Above-mentioned favus of the scalp flower extract has following feature：
The extract is Papaveraceae convex combination algorithm herbaceos perennial slenderstalk dicranostigma herb（Dicranostigma leptopodum
(maxim.）fedde）Herb alcohol extract or water extract.
Application of the above favus of the scalp flower extract as plant antimicrobial, it is characterised in that：Favus of the scalp flower extract contains with wood
It is secondary many with Chelerythrine, sanguinarine, (6aS)-2,10,11-Trimethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g, isocorydine, coptisine and Biflorine etc. based on magnoflorine
Plant active ingredient.
The extraction and separation technology of above slenderstalk dicranostigma herb antibiotic effective ingredient, it is characterised in that：
The extractive technique is related to extract slenderstalk dicranostigma herb vegetable material with alcohols solvent such as ethanol, methyl alcohol etc., using various molten
Agent is extracted to the active ingredient in crude extract, and using chromatographic technique the separation of monomer active ingredient is carried out.
Antibacterial activity in vitro is carried out using mycelial growth rate method to favus of the scalp flower extract, extract and monomeric compound to comment
Valency.By being compared with positive drug, it was demonstrated that favus of the scalp flower extract and its active component involved in the present invention is planted more to
There is significant inhibitory activity in thing pathogen, with being used to prepare the great potential of plant antimicrobial medicine as active ingredient.
Favus of the scalp flower extract and its active component have significant as the application for preparing plants antimicrobial medicine to following phytopathogen
Inhibitory activity：
Apple decay pathogen；Gibberella saubinetii pathogen；Early blight of tomato opportunistic pathogen；Curvularia pathogen；Pumpkin is withered cause of disease
Bacterium；Cotton is withered, withered germ of water-melon；Dry rot of potato opportunistic pathogen；Alternaria brassicae；Tobacco brown spot pathogen；Apple anthrax
Germ；Rice blast fungus；Botryosphaeria berengeriana f. sp, tomato Cercospora Sojina Hara, etc..
Separate below in conjunction with subordinate list, extraction, Structural Identification and determination of activity are described in further detail to the present invention：
First, the preparation of favus of the scalp flower extract and its enrichment of active component：
With 95% industrial alcohol, methyl alcohol or water as solvent, according to 1:15（g/mL）Solid-to-liquid ratio, to slenderstalk dicranostigma herb at 50-80 DEG C
Herb dry powder（By taking 100 g as an example）It is stirred or ultrasonic assistant extraction, continuous extraction 3-5 time, every time 2 h.Extract every time
After end, extract is leached while hot, then plant residue is carried out again with fresh solvent to repeat extraction.Merge all extracts,
It is stored at room temperature overnight, decompression filters sediment.Remove gained filtrate under reduced pressure solvent at 40 DEG C, to bottle green paste extract
The g of thing 24（A）, recovery rate is 24%.
Extract A is dissolved in into the methanol aqueous solution of 100 mL 25%, successively with petroleum ether, chloroform and extracting n-butyl alcohol,
Every kind of solvent continuous extraction three times.The each consumption of petroleum ether is 100 mL, and three consumptions of chloroform are 50 mL, n-butanol three
Secondary consumption is followed successively by 50,40,30 mL.Merge the extract of each same solvent respectively, and remove solvent under reduced pressure, oil is obtained successively
The g of ether extract 2.4（P）, the g of chloroform extract 2.1（C）With the g of n-butyl alcohol extract 9.8（B）.By the water after extracting n-butyl alcohol
Phase evaporated under reduced pressure, obtains the g of the hydrotrope 9.7（H）.
2nd, the separation of active ingredient and identification in favus of the scalp flower extract：
1. in chloroform extract active ingredient separation
1.3 g chloroform extract C are taken, using 3.5 × 28 cm chromatographic columns, 200-300 mesh silica gel, dry method loading carries out column chromatography
Separate.Eluant, eluent is followed successively by chloroform-methanol（10:1; 9:1; 8:1）.F1, F2 are obtained successively（1.05 g）, F3（45.2 mg）, F4
（29.4 mg）And F5（90.8 mg）Part.
Using identical chromatographic column and silica gel, column chromatography for separation is carried out to F1 parts.Eluant, eluent is chlorofonn-ethylacetate
（7:1; 5:1; 3:1; 2:1）, obtain SF1（41 mg）Part.Carry out silica gel thin-layer chromatography separation to SF1, solvent be chloroform-
Ethyl acetate（6:1）, obtain monomeric compound TCH-12（Chelerythrine）（3.3 mg）、TCH-13（Sanguinarine alkali）（2.8
mg）.
F3-F5 are merged, silica gel thin-layer chromatography preparative separation is carried out, solvent is chloroform-methanol（20:1）, obtain list
Body compound TCH-7（(6aS)-2,10,11-Trimethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g）（7.3 mg）And TCH-8（Isocorydine）（6.8 mg）.
2. in n-butyl alcohol extract active ingredient separation
1 g n-butyl alcohol extract B are taken, with 5 mL dichloromethane, after the min of ultrasonication 5, is filtered.Collection insoluble matter, and to
5 mL methyl alcohol are wherein added, after the min of ultrasonication 5, insoluble matter is filtered.Filtrate is filtered with 0.22 micron of ultrafiltration membrance filter, gained
Liquid carries out C18 reversed-phase column column chromatographies, wet method loading.With methyl alcohol-water that polarity is incremented by（10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%）Carry out gradient elution, the mL of 10% -60% eluent each gradient about 100,70% -90% eluent each
The mL of gradient about 500.By 70% methanol-eluted fractions（UV365It is in down yellow fluorescence）Evaporated under reduced pressure, obtains monomeric compound TCH-11
（Coptisine）.
2.4 g n-butyl alcohol extract B are taken, silica gel is carried out（110 g）Pillar layer separation.Chromatographic column specification is 3.5 × 28
Cm, dry method loading.Eluant, eluent is chloroform-methanol（6:1; 4:1; 1:1; 1:2）, obtain BF1-F9 parts.Wherein BF2, BF4,
The quality of BF6, BF7, BF8, BF9 point is than being 91,82,119,509,32,340 mg.
By BF6（119.4 mg）And BF7（509.5 mg）Merge, with LH-20 posts column chromatography is carried out, with methanol-eluted fractions, according to
It is secondary to obtain BF6-1（298 mg）And BF6-2（172 mg）Part.BF6-1 and BF6-2 is merged, silica gel column chromatography, chromatographic column is carried out
Specification is 3.5 × 28 cm, and silica gel is 200-300 mesh, and eluant, eluent is followed successively by chloroform-methanol (6:1; 4:1; 3:1; 2:1),
Obtain 117 mg white solids（TCH-3, Biflorine）.
To BF8 parts（32.3 mg）Thin-layer chromatography preparative separation is carried out, solvent is methanol-water (1:1), collect 365
The material in sky blue spot, obtains 7.6 mg pale yellow powder shape solids under nm（TCH-4, magnoflorine）.
2nd, the physicochemical property of monomeric compound and Spectrum Analysis in slenderstalk dicranostigma herb：
Chelerythrine（TCH-12）, yellow powder is dissolved in chloroform, is soluble in methyl alcohol.Meet Dragendorff's reagent and show orange red.
m.p. 199–200 ℃ (H2O–MeOH); 13C-NMR (125 MHz, CD3OD) δ107.3,151.0,150.9,105.3,
121.8,133.5,152.1,120.1,147.6,151.8,127.6,120.9,130.1,127.2,119.7,63.2,57.8,
132.8,134.3,104.5,53.2.1H-NMR (CDOD3, TMS) δ9.92 (1H, s), 8.60 (1H, d, J=9.0
Hz), 8.56 (1H, d, J=9.0 Hz), 8.10 (1H, d, J=9.0 Hz), 8.08 (1H, s), 8.10 (1H,
D, J=9.0 Hz), 7.49 (1H, s), 6.26 (2H, s), 4.97 (3H, s), 4.27 (3H, s), 4.12 (3H, s);
ESI–MS m/z 348 [M]+.
Sanguinarine（TCH-13）, blood red powder, meet Dragendorff's reagent show orange red.m.p. 244–245 ℃ (H2O–
MeOH); 13C-NMR (125 MHz, CD3OD) δ107.2,150.6,150.6,105.2,121.8,133.0,150.7,
111.1,148.1,149.3,121.4,118.4,128.9,127.4,119.8,133.0,133.9,104.3,106.5,53.2.1H-NMR (CDOD3) δ9.95 (1H, s), 8.57 (1H, d, J=8.9 Hz), 8.48 (1H, d, J=8.8
Hz), 8.19 (1H, d, J=8.9 Hz), 8.13 (1H, s), 7.95 (1H, d, J=8.8 Hz), 7.55 (1H,
S), 6.54 (2H, s), 6.30 (2H, s), 4.49 (3H, s); ESI–MSm/z 332[M]+.
Coptisine（TCH-11）, yellow powder dissolves in DMSO and 50 DEG C of methyl alcohol.In UV365Lower displaing yellow fluorescence, meets bismuth iodide
Potassium reagent shows orange red.13C-NMR (125 MHz, CD3OD) δ150.2,148.1,147.5,145.0,144.2,137.2,
132.7,131.0,122.2,121.5,121.4,120.9,112.0,108.8,105.7,104.9,102.5,55.8,26.7；1H-NMR (CDOD3) δ9.81 (1H, s), 8.83 (1H, s), 7.97 (1H, d,J=8.4), 7.84 (1H, d,J
=8.2 Hz), 7.71 (1H, s), 7.06 (1H, s), 6.49 (2H, s), 6.14 (2H, s), 4.85 (2H, brs),
3.19 (2H, brs)。
Corydaline（TCH-7）, white powder dissolves in chloroform and methyl alcohol and acetone.Meet Dragendorff's reagent and show orange red
。13C-NMR (125 MHz, CDCl3) δ151.8,149.2,143.8,142.3,130.7,127.9,126.4,124.4,
123.9,119.2,111.3,110.8,62.7,62.0,56.0,52.7,43.9,35.4,28.9；1H-NMR (CDCl3) δ
8.72 (1H, s), 7.09 (1H, d,J=8.2 Hz), 6.88 (1H, d,J=8.2 Hz), 6.70 (1H, s),
3.92 (3H, s), 3.91 (3H, s), 3.74 (3H, s), 3.18 (1H, m), 3.06 (1H, d-like,J =
13.3 Hz), 3.04 (1H, d-like,J=13.3 Hz), 2.98 (1H, d-like, J=12.8 Hz), 2.68
(1H, d-like, J=16.0 Hz), 2.56 (3H, s), 2.55 (1H, m), 2.44 (1H, t,J = 13.0
Hz)。
Isocorydine（TCH-10）, colourless powder dissolves in chloroform, methyl alcohol, in UV365Aobvious blue-fluorescence down.Meet iodate
Bismuth potassium reagent shows orange red.13C-NMR (125 MHz, CD3OD) δ151.7,149.1,143.3,142.4,129.8,
129.7,128.0,125.3,119.8,119.1,111.5,111.2,62.9,60.9,55.3,55.0,52.3,42.4,
34.9,28.2；1H-NMR (CDOD3) δ 6.93 (1H, d, J = 8.1), 6.87 (1H, d,J = 8.1), 6.86
(1H, s), 3.89 (3H, s), 3.87 (3H, s), 3.65 (3H, s), 3.10 (1H, dd, J=3.5,13.0,
Hz), 3.10 (1H, dd,J = 13.2,3.4, Hz), 3.06 (1H, dd, J=6.0,11.5 Hz), 2.85
(1H, d-like, J=13.0 Hz), 2.76 (1H, dd,J = 16.6,3.6 Hz), 2.54 (3H, s), 2.51
(1H, 2 × t, J=4.0,12.0 Hz), 2.38 (1H, t, J=13.0 Hz).
Biflorine（TCH-3）, white powder dissolves in chloroform, methyl alcohol.Meet Dragendorff's reagent and show orange red.1H
NMR (400 MHz,CDCl3)δ6.91 (s, 1H), 6.65 (1H, s), 2.56(4H, br s), 1.92 (3H, s),
3.59 (2H, br s), 6.67 (1H, d, J = 8.0 Hz), 6.68 (1H, d, J = 8.0 Hz), 3.92
(2H, br s), 5.95 (2H, s), 5.93 (2H, s); 13C NMR (100 MHz, CDCl3)δ194.9, 148.0,
146.3, 146.0, 145.9, 136.1 132.7, 128.9, 125.0, 117.8, 110.5, 108.1, 106.7,
100.8, 101.2, 57.7, 50.8, 46.4, 41.4, 31.8. EI-MS m/z 353 [M]+, 338, 309,
281, 267, 252, 223, 209, 190, 163, 148, 134, 91.
Magnoflorine（TCH-4）, pale yellow powder is dissolved in chloroform, is slightly soluble in methyl alcohol, in UV365Aobvious blue-fluorescence down, with iodate
Bismuth potassium reagent shows punctation.
1H NMR (500 MHz, CDOD3)δ6.64 (1H, d, J=8.0 Hz), 6.35 (1H, s), 6.42
(1H, d, J=8.0 Hz), 3.85 (3H, s), 3.75 (3H, s), 3.62 (1H, br d, J=14.3 Hz),
3.20 (1H, m), 3.11 (3H, s), 3.02 (2H, m), 2.82 (1H, d, J=10.0 Hz), 2.65 (3H,
S), 2.43 (1H, d-like, J=13.0 Hz), 2.25 (1H, t, J=13.0 Hz);13C NMR (125 MHz,
CDOD3)δ152.8,151.6,150.7,149.8,126.1,123.4,123.3,117.0,115.8,121.1,110.4,
109.3,70.8,62.1,55.0,54.7,53.8,43.6,31.6,24.6；
3rd,（E）The antibacterial activity of-cinnamate derivative compound：
1st, for examination bacterium：
Fusarium graminearum（Fusariumg graminearum）, rice blast fungus（Pyricularia grisea）, corn
Curved spore germ（Curvularia lunata）, Valsa mali（Valsa mali）, tomato early blight bacterium（Alternaria solani）, cotton-wilt fusarium（Fusarium oxysporium f. sp. Vasinfectum）, withered germ of water-melon
（Fusarium oxysporium f. sp. Niveum）, dry rot of potato bacterium（Fusarium solani）, Chinese cabbage blackspot
Germ（Alternaria brassicae）, tobacco brown spot pathogen（Alternaria alternate）, pumpkin wilt
（Mycosphaerella melonis）, botrytis cinerea（Botrytis cinerea）.
2nd, sample preparation：
40 mg crude extracts or extract, or the monomeric compound of 10.0 mg are weighed, the DMSO of 10 mL 5% is dissolved in（v/v）,
It is standby after being sterilized 30 minutes with ultra violet lamp on super-clean bench.
3rd, the preparation of PDA culture medium：
By the g of peeled potatoes 200.0 choppings, 1000 mL water are added, boil 30 min, filtered through gauze, filtrate water constant volume is arrived
1000 mL, after adding 20.0 g glucose, 15.0 g agar powders, heating it is all dissolved, are divided in triangular flask.
4th, the measure of antibacterial activity：
Antibacterial activity is carried out using suppression mycelial growth rate method.By 10 mL test liquids of above-mentioned preparation, 190 mL sterilizings are poured into
Afterwards in the PDA culture medium in molten state, pour into while hot in 13 sterile petri dish after quick mixing, obtain final product containing 50μG/mL is supplied
The culture medium of examination monomeric compound or 2.0 mg/mL extracts；Using the PDA culture medium containing 0.25% DMSO as blank.
After culture medium solidifying, with the card punch of a diameter of 0.5 cm from for trying to produce bacterium where bacterium colony edge mycelial growth is vigorous
Cake, is carefully placed in bacteria cake above pastille culture medium with transfer needle.The mycelia of bacteria cake faces down.Per three bacteria cakes of ware, with three
The angular point of central authorities for being put in culture dish, after then adding a cover and marking, are placed in the incubator of 25 DEG C and 80% humidity and cultivate.Each
Process in triplicate.After cultivating 72 h, take crossing method to measure colony diameter, take its mean value, calculate by formula below
The bacteriostasis rate of each test compound.
Bacterium colony extends diameter/cm=colony diameter mean value 0.5（Bacteria cake diameter）
Slenderstalk dicranostigma herb alcohol extracts and each solvent extract（2 mg/mL）Result of the test be listed in the table below.As a result show, slenderstalk dicranostigma herb
Alcohol extracts show different degrees of inhibitory activity to all for examination bacterium, and its active component is concentrated mainly on chloroform extraction
In taking thing and n-butyl alcohol extract.
7 kinds of monomeric compounds in slenderstalk dicranostigma herb（50μg/mL）The result of the test of 12 kinds of plant pathogenic fungis is listed in the table below.
The title and numbering of each compound is respectively：Chelerythrine（TCH-12）, sanguinarine（TCH-13）, coptisine（TCH-11）,
(6aS)-2,10,11-Trimethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g（TCH-7）, isocorydine（TCH-10）, Biflorine（TCH-3）, magnoflorine（TCH-4）.
As a result show, Chelerythrine（TCH-12）, sanguinarine（TCH-13）And magnoflorine（TCH-4）To 12 kinds of plants
Disease fungus is respectively provided with stronger inhibitory activity, and to the activity of indivedual bacterium positive drug probenazole is better than.
Toxicity equation and medium effective concentration of the magnoflorine to 7 kinds of phytopathogens（EC50）Value is listed in the table below.As a result table
It is bright, EC of the magnoflorine to 7 kinds of phytopathogens50For 10-38 μ g/ml, average EC50For 23.4 μ g/ml, with directly as
Plants antimicrobial active ingredient is used to prepare the potentiality of plants antimicrobial medicine.
Fig. 1 is the high-efficient liquid phase chromatogram of slenderstalk dicranostigma herb alcohol extracts.As a result show, slenderstalk dicranostigma herb alcohol extracts it is main
Composition is isocorydine, magnoflorine and coptisine, content of the three in slenderstalk dicranostigma herb is respectively 1.14%, 0.2%,
0.095%.The content of other compositions is less.
The activity and its content in slenderstalk dicranostigma herb of comprehensive each monomeric compound, it may be determined that main antibacterial in slenderstalk dicranostigma herb
Referred to as magnoflorine.
As can be seen here, favus of the scalp flower extract according to the present invention and its active ingredient have wider resisting to phytopathogen
Bacterium is composed and stronger antibacterial activity, with being used to prepare the potential use of plants antimicrobial medicine, can be used as plants antimicrobial medicine
Active ingredient.
Present disclosure is not limited to cited by embodiment, and those of ordinary skill in the art are by reading description of the invention
And any equivalent conversion that technical solution of the present invention is taken, it is claim of the invention and is covered.
Claims (7)
1. favus of the scalp flower extract, it is characterised in that：
The extract is Papaveraceae（Papaveraceae）Convex combination algorithm（Dicranostigma）Herbaceos perennial slenderstalk dicranostigma herb
（Dicranostigma leptopodum (maxim.）fedde）Herb or aerial part alcohol extract or water extract.
2. favus of the scalp flower extract according to claim 1, it is characterised in that：
Include antimicrobial component in the favus of the scalp flower extract, including magnoflorine, Chelerythrine, sanguinarine, (6aS)-2,10,11-Trimethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g,
Isocorydine, coptisine and Biflorine.
3. the application of favus of the scalp flower extract as claimed in claim 1, it is characterised in that：
The extract is used as the application for preparing plants antimicrobial medicine.
4. the application of favus of the scalp flower extract according to claim 3, it is characterised in that：
The extract has significant inhibitory activity to following phytopathogen：
Apple decay pathogen；Gibberella saubinetii pathogen；Early blight of tomato opportunistic pathogen；Curvularia pathogen；Pumpkin is withered cause of disease
Bacterium；Cotton is withered, withered germ of water-melon；Dry rot of potato opportunistic pathogen；Alternaria brassicae；Tobacco brown spot pathogen；Apple anthrax
Germ；Rice blast fungus；Botryosphaeria berengeriana f. sp；Tomato Cercospora Sojina Hara.
5. the extracting method of favus of the scalp flower extract as claimed in claim 1, it is characterised in that：
Comprise the following steps：
Step one：With ethanol, methyl alcohol or water as solvent, slenderstalk dicranostigma herb vegetable material is carried out heating extraction, merge what is repeatedly extracted
Extract, is stored at room temperature overnight, and decompression filters sediment, and gained filtrate decompression is evaporated off into solvent, obtains extract；
Step 2：Methanol aqueous solution is dissolved in by extract obtained, oil is used with petroleum ether, chloroform and n-butanol or successively successively
Ether, n-butanol are extracted；The n-butyl alcohol extract obtained by chloroform extract and n-butyl alcohol extract or rear method obtained by front method
As favus of the scalp flower extract.
6. the extracting method of favus of the scalp flower extract according to claim 5, it is characterised in that：
In step：
Slenderstalk dicranostigma herb vegetable material is 1 g with the solid-liquid mixing ratio of solvent:15 mL, solution temperature is 50-80 DEG C, is stirred during dissolving
Or ultrasonic assistant；
Temperature when extract removes solvent under reduced pressure is 30～40 DEG C.
7. the extracting method of favus of the scalp flower extract according to claim 5, it is characterised in that：
In step 2：
The extract of slenderstalk dicranostigma herb vegetable material is dissolved in methanol-water identical in quality with vegetable material, volume fraction is 10%～25%
In solution, then successively with petroleum ether, chloroform and n-butanol or petroleum ether, extracting n-butyl alcohol, every kind of solvent extraction three times；
During petroleum ether extraction, each solvent load is identical with the volume for being extracted liquid；
During chloroform extraction, each solvent load is the half for being extracted liquid product；
During extracting n-butyl alcohol, three solvent loads are followed successively by and are extracted 50%, 40% and the 30% of liquid product.
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| CN111560016A (en) * | 2020-05-11 | 2020-08-21 | 郑州铁路职业技术学院 | Method for extracting and separating coptisine from celandine |
| CN112021343A (en) * | 2020-09-27 | 2020-12-04 | 浙江农林大学 | Plant source fumigant suitable for grain storage and preparation method thereof |
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| CN110780020A (en) * | 2019-12-06 | 2020-02-11 | 郑州铁路职业技术学院 | Detection method for simultaneously determining ten alkaloids in tinea capitis flowers by HPLC |
| CN111560016A (en) * | 2020-05-11 | 2020-08-21 | 郑州铁路职业技术学院 | Method for extracting and separating coptisine from celandine |
| CN112021343A (en) * | 2020-09-27 | 2020-12-04 | 浙江农林大学 | Plant source fumigant suitable for grain storage and preparation method thereof |
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