CN105906566B - A kind of alkaloid compound, preparation method and use with antibacterial activity in tobacco - Google Patents
A kind of alkaloid compound, preparation method and use with antibacterial activity in tobacco Download PDFInfo
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- CN105906566B CN105906566B CN201610338953.8A CN201610338953A CN105906566B CN 105906566 B CN105906566 B CN 105906566B CN 201610338953 A CN201610338953 A CN 201610338953A CN 105906566 B CN105906566 B CN 105906566B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
- C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
- C07D221/04—Ortho- or peri-condensed ring systems
- C07D221/06—Ring systems of three rings
- C07D221/14—Aza-phenalenes, e.g. 1,8-naphthalimide
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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
- A01N43/42—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 condensed with carbocyclic rings
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- D—TEXTILES; PAPER
- D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
- D21H—PULP COMPOSITIONS; PREPARATION THEREOF NOT COVERED BY SUBCLASSES D21C OR D21D; IMPREGNATING OR COATING OF PAPER; TREATMENT OF FINISHED PAPER NOT COVERED BY CLASS B31 OR SUBCLASS D21G; PAPER NOT OTHERWISE PROVIDED FOR
- D21H27/00—Special paper not otherwise provided for, e.g. made by multi-step processes
- D21H27/10—Packing paper
Abstract
The invention discloses a kind of alkaloid compound, chemical name is 8 hydroxyl, 3 methylol 6,9 dimethyl 7H benzos [de] isoquinolin, 7 ketone, molecular formula C15H13NO3And there are following structures：The invention also discloses the methods that above-mentioned alkaloid compound is prepared from tobacco.The invention also discloses the purposes of above compound, show it with preferable bacteriostasis through active testing.The compounds of this invention structure novel, and there is preferable antibacterial activity.The compound is used for cigarette tipping paper, can eliminate or reduce the possibility of bacteria breed and breeding in cigarette tipping paper.
Description
Technical field
The invention belongs to technical field of tobacco chemistry, and in particular to a kind of alkaloids extracted for the first time from tobacco
Compound.Meanwhile the application the invention further relates to the preparation method of the compound and in antibiotic package paper.
Background technology
Tobacco is the plant that chemical composition is the most complicated in the world, and secondary metabolite is very abundant, according to nineteen eighty-two Dube
With the reports such as Green, the chemical composition identified in tobacco is just more than 2549 kinds, by 2008, Rodgman and perfetti
According to the report, the compound sum found in tobacco, tobacco and cigarette smoke is about 8700 kinds.Currently, people
The monomer chemistries substance come is identified from tobacco just more than more than 3000 kind, but also there are many ingredients not yet to identify.Cigarette
Grass can also therefrom extract a variety of chemical compositions for having utility value in addition to being mainly used for cigarette smoking purposes, therefrom find have out
Send out the guiding compound of utility value.Alkaloid (alkaloid) is that the one kind being present in organism (predominantly plant) contains
The alkaline organic compound of nitrogen, it is most of to there is complicated cyclic structure, nitrogen to be included in ring more, there is significant bioactivity,
It is the important sources of drug and biological pesticide.To make full use of our province tobacco resources advantage, it is natural further to find new activity
Product, we study tobacco components, and isolated a kind of new alkaloid from Yunnan Flue-cured Tobacco tobacco leaf
Relevant report is not yet seen in class compound, the compound, it is worth mentioning at this point that the compound has significant antibacterial activity.
Invention content
The purpose of the present invention is to provide a kind of new alkaloid compounds.
It is a further object to provide a kind of methods preparing the alkaloid compound.
The present invention also aims to provide application of the alkaloid compound in antibiotic package paper.
Unless otherwise indicated, the percentage employed in the present invention is mass percent.
The present invention isolates a kind of new alkaloid compound, molecular formula C from tobacco15H13NO3, have following
Structural formula：
The compound is named as：8- hydroxyls -3- methylols -6,9- dimethyl -7H- benzos [de] isoquinolin -7- ketone, English
Literary fame is：8-hydroxy-3-(hydroxymethyl)-6,9-dimethyl-7H-benzo[de]quinolin-7-one.
The method for preparing the alkaloid compound, this approach includes the following steps：
(1) medicinal extract extracts：Tobacco sample is crushed, with high concentration methanol (w%：80%~100%) or high concentration ethanol
(w%：80%~100%) or high concentration acetone (w%：60%~90%) it is Extraction solvent, Extraction solvent：Tobacco (weight ratio)
=2~4:1, for 24 hours~72h is impregnated, is extracted 3~5 times, merges extracting solution, filtering and concentrating into medicinal extract；
(2) silica gel column chromatography：Medicinal extract is used after being dissolved with the pure methanol of 1.5~3 times of amounts of weight ratio or straight alcohol or pure acetone
80~100 mesh silica gel mixed samples of 0.8~1.2 times of weight ratio, with 160~300 mesh silica gel dry column-packings of 2~4 times of amounts of weight ratio
Carry out silica gel column chromatography；With volume proportion for (1:0、20:1、9:1、8:2、7:3、6:4、1:1 and 1:2) chloroform-acetone solution
Gradient elution is carried out, identical part is merged, collect each section eluent and is concentrated；
(3) high pressure liquid chromatography isolates and purifies：The 8 of column chromatography eluent:2 parts are further detached with high pressure liquid chromatography
It purifies up to the alkaloid compound.
It is to use 21.2mm × 250mm, 5 μm of C that high pressure liquid chromatography, which isolates and purifies,18Chromatographic column, flow velocity 20mL/min,
The methanol that mobile phase is 52%, UV detector Detection wavelength are 342nm, and 200 μ L of each sample introduction collect the chromatography of 31.5min
Peak is evaporated after repeatedly adding up.
After purification through the high performance liquid chromatography separation, a preferred subsequent process scheme is that gained compound is again
It is dissolved with pure methanol, then using pure methanol as mobile phase, is detached with gel filtration chromatography, further to isolate and purify.
1 compound 1 of table1H-NMR and13C-NMR data (500/125MHz, C5D5N)
The structure for the alkaloid compound that method described above is prepared is measured by the following method.Of the present inventionization
Conjunction object is orange red jelly, and HR-ESI-MS shows that its quasi-molecular ion peak is 278.0798 [M+Na]+, in conjunction with1H- and13C-
H NMR spectroscopy determines that molecular formula is C15H13NO3.Its infrared spectrum, which is shown in compound, hydroxyl (3412cm-1), carbonyl (1650cm-1) and aromatic ring (1610,1567,1462cm-1) signal, ultraviolet spectra has absorption maximum to also confirm that compound in 246 and 342nm
In there are aromatic ring structures.From1H H NMR spectroscopies (attribution data is shown in Table -1) signal, which can be seen that in compound, 3 aromatic series hydrogen letters
Number (δH8.48s, 8.01 (d) 8.1,7.67 (d) 8.1)；2 methyl signals (δH2.51s and 2.80s)；The methylene of 1 oxidation
Base (δH4.62)；And 1 phenolic hydroxyl group (δH10.42).Compound13C NMR and DEPT spectrums show in compound there is 8 virtues
Fragrant quaternary carbon (δC125.3s、131.6s、146.6s、124.7s、151.2s、123.0s、158.7s、120.2s)；3 fragrance
Methyl (δC142.6d、128.0d、134.3d)；1 alpha, beta-unsaturated carbonyl (δC181.1s)；2 methyl (δC10.5q and
24.8q)；And methylene (the δ of 1 oxidationC66.9t).The corresponding carbon signal of all hydrogen is belonged to according to hsqc spectrum.Pass through
H3- 13 and C-5, C-6, C-6a, H-5 and C-3a, C-6a, C-14, H-4 and C-3, C-3a, C-11, H2- 12 and C-2, C-3, C-
3a and H-2 is related to the HMBC of C-3, C-3a, C-12, C-10, it can be verified that there are 3- methylol -6- methyl is different in compound
The structure fragment of Kui quinoline.According to the degree of unsaturation 10 of compound, should also there be 1 ring in the compound.According to H3- 14 and C-8, C-
9, the HMBC of C-10 is related, can speculate α, and the positions C-10 of β-carbon (C-9) and isoquinolin ring in beta-unsaturated carbonyl are connected, and
Carbonyl (C-7) should be connected to form 1 hexatomic ring with C-6a.Another 1 methyl (C-14) is substituted in C-9 by H3- 14 and C-8,
The HMBC correlations of C-9, C-10 are determined.Phenolic hydroxyl group is substituted in C-8 can be according to the chemical displacement value (δ of C-8C151.2) exist
Low field and phenolic hydroxyl group signal (δH10.42) related to the HMBC of C-7, C-8, C-9 to determine.So far the structure of this compound
It is determined.The Compound nomenclature is：8- hydroxyls -3- methylols -6,9- dimethyl -7H- benzos [de] isoquinolin -7- ketone.
Antimicrobial Screening is carried out to the compound, to staphylococcus aureus, Escherichia coli, angstrom uncommon bacterium, withered grass
Bacillus, proteus etc. have significant activity.
The compound is applied in cigarette tipping paper to compare with control, and the tipping paper detection for adding this compound is thin
Bacterium sum, coliform, staphylococcus aureus, Pseudomonas aeruginosa, hemolytic streptococcus, total number of fungi substantially reduce；To large intestine
Bacillus (ATCC25922), staphylococcus aureus (ATCC6538) bacteriostasis rate entirely reach 96% or more, can reduce or disappear
Except the cigarette tipping paper and during storage possibility of bacteria breed and breeding, in addition, in smoking property of cigarette, transmittance process,
The antibacterial action also can play inhibiting effect to the contaminated microorganism of tipping paper on Cigarette.
Compared with prior art, the present invention has the advantages that following prominent：(1) the compound of the present invention raw material is easy to get, extraction
Method is simple, can be easily separated to obtain；Molecular structure is also simple, easy to implement artificial synthesized.(2) conventional column chromatography and height are used
The preparation method that effect liquid phase chromatogram combines, compound preparation manipulation flow is simple, and the compounds of this invention purity obtained is high, after
Continuous industrialized production is easy to implement.(3) the compounds of this invention is nontoxic to animal, safe to use, shows good antibacterial and lives
Property, 96% or more is entirely reached to the bacteriostasis rate of Escherichia coli, staphylococcus aureus etc.；It, can applied to cigarette tipping paper
Inhibiting effect is played to the contaminated microorganism of cigarette tipping paper.Cigarette tipping paper is directly being rolled up with oral contact, the compound
Use in cigarette tipping paper can avoid cigarette suck, microbial contamination in transmittance process, effectively increase defending for cigarette
Raw and safety.
Description of the drawings
Fig. 1 is the carbon-13 nmr spectra of alkaloid compound of the present invention；
Fig. 2 is the nuclear magnetic resonance spectroscopy of alkaloid compound of the present invention；
Fig. 3 is that the main HMBC of alkaloid compound of the present invention is related.
Specific implementation mode
The present invention is described in further detail with reference to the accompanying drawings and examples, but not in any way to the present invention
Limit, be based on present invention teach that made by any transformation or improvement, each fall within protection scope of the present invention.
Embodiment 1
Prepare alkaloid compound C15H13NO3, including medicinal extract extraction, silica gel column chromatography, high pressure liquid chromatography separation, tool
Body uses following steps：
1. medicinal extract extracts：Tobacco leaf is taken to crush, with high concentration methanol (w%：Or high concentration ethanol (w% 95%)：95%) or
High concentration acetone (w%：70%) it is Extraction solvent, Extraction solvent：Tobacco (weight ratio)=3:5,54h is impregnated, is extracted 4 times, is closed
And extracting solution, filtering and concentrating are at medicinal extract.
2. silica gel column chromatography：Medicinal extract uses weight after being dissolved with the pure methanol of 2.5 times of amounts of weight ratio or straight alcohol or pure acetone
80~100 mesh silica gel mixed samples than 1.2 times carry out silica gel column chromatography with 250 mesh silica gel dry column-packings of 3 times of amounts of weight ratio；With
Volume proportion is (1:0、20:1、9:1、8:2、7:3、6:4、1:1 and 1:2) chloroform-acetone solution carries out gradient elution, merges
Identical part is collected each section eluent and is concentrated.
3. high pressure liquid chromatography detaches：The 8 of column chromatography eluent:2 parts are further isolated and purified with high pressure liquid chromatography
Up to the alkaloid compound, it is to use 21.2mm × 250mm, 5 μm of C that high pressure liquid chromatography, which isolates and purifies,18Chromatography
Column, flow velocity 20mL/min, mobile phase be 52% methanol, UV detector Detection wavelength be 342nm, 200 μ L of each sample introduction,
The chromatographic peak of 31.5min is collected, is evaporated after repeatedly adding up.
High pressure lipuid chromatography (HPLC) isolate and purify after substance, a preferred post-processing scheme is that gained compound is again
It is dissolved with pure methanol, then using pure methanol as mobile phase, is detached with gel filtration chromatography, further to isolate and purify.
Raw tobacco material used in the present invention is not limited by area and kind, the present invention may be implemented, below to derive from cloud
The raw tobacco material of southern different sources, the present invention will be further described：
Embodiment 2
Tobacco sample derives from Yunnan Yuxi, and kind is Yuxi K326.Tobacco sampling 2.0kg is crushed with 95% methanol
Extraction 5 times, for 24 hours, extracting solution merges for extraction every time, and filtering is concentrated under reduced pressure into medicinal extract, obtains medicinal extract 105g.Medicinal extract weight ratio 2.0
The thick silica gel mixed sample of 100 mesh of 120g, the 160 mesh silica gel of 0.6kg is used to fill column progress silica gel column chromatography after the pure methanol dissolving of amount again,
It is 1 with volume proportion:0、20:1、9:1、8:2、7:3、6:4、1:1、1:2 chloroform-acetone gradient elution, TLC monitorings merge phase
Same part obtains 8 parts, and wherein volume proportion is 8:2 chloroform-acetone elution fraction prompt logical sequence 1,100 half of peace prepares high
Effect liquid phase chromatogram detaches, and using 52% methanol as mobile phase, it is solid that Zorbax SB-C18 (21.2 × 250mm, 5 μm), which prepare column,
Determine phase, flow velocity 20ml/min, UV detector Detection wavelength is 342nm, and 200 μ L of each sample introduction collect the chromatography of 31.5min
Peak is evaporated after repeatedly adding up；Products therefrom is dissolved with pure methanol again, then using pure methanol as mobile phase, with Sephadex LH-
20 gel filtration chromatographies detach to get the noval chemical compound.
Embodiment 3
Tobacco sample derives from Dali, and kind is cloud and mist 200, by tobacco sampling 3.5kg choppings, with 95% ethyl alcohol
48h is extracted in extraction 4 times every time, and extracting solution merges, and filtering is concentrated under reduced pressure into medicinal extract, obtains medicinal extract 250g.Medicinal extract weight ratio 2.0
The thick silica gel mixed sample of 80 mesh of 250g, the 200 mesh silica gel of 1.2kg is used to fill column progress silica gel column chromatography after the pure methanol dissolving of amount again,
It is 1 with volume proportion:0、20:1、9:1、8:2、7:3、6:4、1:1、1:2 chloroform-acetone gradient elution, TLC monitorings merge phase
Same part obtains 8 parts, and wherein volume proportion is 8:2 chloroform-acetone elution fraction prompt logical sequence 1,100 half of peace prepares high
Effect liquid phase chromatogram detaches, and using 52% methanol as mobile phase, it is solid that Zorbax SB-C18 (21.2 × 250mm, 5 μm), which prepare column,
Determine phase, flow velocity 20ml/min, UV detector Detection wavelength is 342nm, and 200 μ L of each sample introduction collect the chromatography of 31.5min
Peak is evaporated after repeatedly adding up；Products therefrom is dissolved with pure methanol again, then using pure methanol as mobile phase, with Sephadex LH-
20 gel filtration chromatographies detach to get the noval chemical compound.
Embodiment 4
Tobacco sample derives from Kunming, Yunnan, and kind is the big gold dollar of safflower, tobacco sampling 5kg is crushed, with the third of 75%
Ketone ultrasonic extraction 3 times extracts 72h every time, and extracting solution merges, and filtering is concentrated under reduced pressure into medicinal extract, obtains medicinal extract 380g.Medicinal extract is used
The thick silica gel mixed sample of 90 mesh of 400g, the 180 mesh silica gel dress column of 2.4kg is used to carry out silicon after the pure methanol dissolving of 1.6 times of amounts of weight ratio
Plastic column chromatography is 1 with volume proportion:0、20:1、9:1、8:2、7:3、6:4、1:1、1:2 chloroform-acetone gradient elution, TLC
Monitoring merges identical part, obtains 8 parts, and wherein volume proportion is 8:2 chloroform-acetone elution fraction An Jielun
1100 half preparative high-performance liquid chromatographics detach, using 52% methanol as mobile phase, Zorbax SB-C18 (21.2 × 250mm, 5 μ
M) it is stationary phase to prepare column, and flow velocity 20ml/min, UV detector Detection wavelength is 342nm, 200 μ L of each sample introduction, is collected
The chromatographic peak of 31.5min is evaporated after repeatedly adding up；Products therefrom is dissolved with pure methanol again, then using pure methanol as mobile phase, is used
Sephadex LH-20 gel filtration chromatographies detach to get the noval chemical compound.
Embodiment 5
The identification of --- --- compound structure
Compound prepared by Example 1-4, the structure for the alkaloid compound that method described above is prepared pass through
Following methods are measured.Compound is orange red jelly, and HR-ESI-MS shows that its quasi-molecular ion peak is 278.0798 [M
+Na]+, in conjunction with1H- and13C-NMR spectrums determine that molecular formula is C15H13NO3.Its infrared spectrum, which is shown in compound, hydroxyl
(3412cm-1), carbonyl (1650cm-1) and aromatic ring (1610,1567,1462cm-1) signal, ultraviolet spectra has in 246 and 342nm
Absorption maximum also confirms that there are aromatic ring structures in compound.From1H H NMR spectroscopies (attribution data is shown in Table -1) signal can be seen that chemical combination
There are 3 aromatic series hydrogen signal (δ in objectH8.48s, 8.01 (d) 8.1,7.67 (d) 8.1)；2 methyl signals (δH2.51s and
2.80s)；Methylene (the δ of 1 oxidationH4.62)；And 1 phenolic hydroxyl group (δH10.42).Compound13C NMR and DEPT
Spectrum, which is shown in compound, 8 fragrant quaternary carbon (δC 125.3s、131.6s、146.6s、124.7s、151.2s、123.0s、
158.7s、120.2s)；3 fragrant methine (δC142.6d、128.0d、134.3d)；1 alpha, beta-unsaturated carbonyl (δC
181.1s)；2 methyl (δC10.5q and 24.8q)；And methylene (the δ of 1 oxidationC66.9t).Belonged to according to hsqc spectrum
All hydrogen corresponding carbon signals.Pass through H3- 13 and C-5, C-6, C-6a, H-5 and C-3a, C-6a, C-14, H-4 and C-3, C-
3a, C-11, H2- 12 and C-2, C-3, C-3a and H-2 it is related to the HMBC of C-3, C-3a, C-12, C-10, it can be verified that chemical combination
There are the structure fragments of the different Kui quinoline of 3- methylol -6- methyl in object.According to the degree of unsaturation 10 of compound, also answered in the compound
There is 1 ring.According to H3- 14 is related to the HMBC of C-8, C-9, C-10, can speculate α, β-carbon (C-9) in beta-unsaturated carbonyl and
The positions C-10 of isoquinolin ring are connected, and carbonyl (C-7) should be connected to form 1 hexatomic ring with C-6a.Another 1 methyl (C-14)
C-9 are substituted in by H3- 14 related to the HMBC of C-8, C-9, C-10 are determined.Phenolic hydroxyl group is substituted in C-8 can be according to C-8
Chemical displacement value (δC151.2) in low field and phenolic hydroxyl group signal (δH10.42) related to the HMBC of C-7, C-8, C-9
It determines.So far the structure of this compound is determined.The Compound nomenclature is：8- hydroxyl -3- methylol -6,9- dimethyl -7H-
Benzo [de] isoquinolin -7- ketone.
Embodiment 6
Compound prepared by Example 3 is yellow jelly.Assay method is same as Example 5, confirms embodiment 3
The compound of preparation is the alkaloid compound --- 8- hydroxyl -3- methylol -6,9- dimethyl -7H- benzos [de]
Isoquinolin -7- ketone.
Embodiment 7
Compound prepared by Example 4 is yellow jelly.Assay method is same as Example 5, confirms embodiment 4
The compound of preparation is described 8- hydroxyls -3- methylols -6,9- dimethyl -7H- benzos [de] isoquinolin -7- ketone.
Embodiment 8
Prepared by Example 1-4 appoints alkaloid compound to carry out antibacterial activity experiment, and test situation is as follows：
Antimicrobial test is carried out with agar diffusion method, and tested bacterium is equably coated in plain agar culture medium (ox first
Meat extract, peptone, sodium chloride, serum, agar) tablet on, then by untested compound (alkaloid compound 10mL DMSO
Dissolving, is diluted with water into the solution of 50 μ g/mL) soaked tablet (diameter 5mm) is placed on the culture medium to carry disease germs, it is put into constant temperature
In case, inhibition zone size is observed after being incubated 24-72h in 25 DEG C.The result shows that：The compounds of this invention to staphylococcus aureus,
Escherichia coli, angstrom uncommon bacterium, hay bacillus, proteus etc. have very strong activity；Inhibiting rate is more than 96%.
Embodiment 9
Safety evaluatio has been carried out to the compounds of this invention, has passed through Micronuclei In The Mouse Bone Marrow experiment, Ames experiments and TK genes
Mutating experiment, it was demonstrated that the compounds of this invention is nontoxic to animal, safe to use.This compound is added to cigarette with the concentration of 50 μ g/mL
On tipping paper；By the People's Republic of China (PRC)《Disposable Sanitary Accessory sanitary standard》The detection method of GB15979-2002,
Take the volume cigarette tipping paper for adding the compounds of this invention, 2.0 × 3.0mm sizes, detection bacterium sum, coliform, golden yellow
Staphylococcus, Pseudomonas aeruginosa, hemolytic streptococcus, total number of fungi.The result shows that adding the tipping paper bacterium of the compounds of this invention
It falls sum to significantly reduce, this compound has obvious inhibiting effect to the bacterium of several tests, to Escherichia coli, golden yellow grape
The bacteriostasis rate of coccus etc. entirely reaches 96% or more.
Claims (5)
1. one kind having the following structural formula alkaloid compound：
The compound is named as：8- hydroxyls -3- methylols -6,9- dimethyl -7H- benzos [de] isoquinolin -7- ketone.
2. a kind of method preparing alkaloid compound described in claim 1, this approach includes the following steps：
(1) medicinal extract extracts:Using tobacco leaf as raw material, tobacco leaf is crushed or is cut into segment, with concentration expressed in percentage by weight 80%~100%
Methanol or ethyl alcohol or the acetone of concentration expressed in percentage by weight 60%~90% are Extraction solvent, Extraction solvent：Weight ratio=2 of tobacco leaf
~4:1, for 24 hours~72h is impregnated, is extracted 3~5 times, merges extracting solution, filtering and concentrating into medicinal extract；
(2) silica gel column chromatography：Medicinal extract carries out silica gel column chromatography with 160~300 mesh silica gel dry column-packings of 2~4 times of amounts of weight ratio；
With volume proportion for 1:0、20:1、9:1、8:2、7:3、6:4、1:1 and 1:2 chloroform-acetone solution carries out gradient elution, merges
Identical part is collected each section eluent and is concentrated；
(3) high pressure liquid chromatography isolates and purifies：The 8 of eluent:2 parts are further isolated and purified with high pressure liquid chromatography up to institute
The alkaloid compound needed.
3. preparation method according to claim 2, it is characterised in that：In step (3), after high pressure liquid chromatography isolates and purifies
Compound dissolved again with pure methanol, then using pure methanol as mobile phase, detached with gel filtration chromatography, it is pure further to detach
Change.
4. preparation method according to claim 2, it is characterised in that：In step (3), high pressure liquid chromatography separation
Purifying is to use 21.2mm × 250mm, 5 μm of C18Chromatographic column, flow velocity 20mL/min, the methanol that mobile phase is 52% are ultraviolet
Detector Detection wavelength is 342nm, and 200 μ L of each sample introduction collect the chromatographic peak of 31.5min, is evaporated after repeatedly adding up.
5. preparation method according to claim 2, it is characterised in that：In step (2), the medicinal extract is through silica gel column layer
Before analysing rough segmentation, after the pure methanol or straight alcohol of 1.5~3 times of weight ratio amount or pure acetone dissolving, with 0.8~1.2 times of weight ratio
80~100 mesh silica gel mixed samples.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140039184A1 (en) * | 2011-02-24 | 2014-02-06 | Wolfgang Bäumler | Phenalene-1-one derivatives, method for producing same and use thereof |
| CN104761526A (en) * | 2015-04-11 | 2015-07-08 | 云南中烟工业有限责任公司 | Isoflavone compound with anti-virus activity as well as preparation method and application thereof |
| CN105481818A (en) * | 2015-11-20 | 2016-04-13 | 云南中烟工业有限责任公司 | Aroma-enhancing moisture-retaining isocoumarin compound, and preparation method and application thereof |
-
2016
- 2016-05-19 CN CN201610338953.8A patent/CN105906566B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140039184A1 (en) * | 2011-02-24 | 2014-02-06 | Wolfgang Bäumler | Phenalene-1-one derivatives, method for producing same and use thereof |
| CN104761526A (en) * | 2015-04-11 | 2015-07-08 | 云南中烟工业有限责任公司 | Isoflavone compound with anti-virus activity as well as preparation method and application thereof |
| CN105481818A (en) * | 2015-11-20 | 2016-04-13 | 云南中烟工业有限责任公司 | Aroma-enhancing moisture-retaining isocoumarin compound, and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| Sesquiterpenes and Alkaloids from the Roots of Alangium chinense;Yan Zhang等;《J. Nat. Prod.》;20130604;第76卷;1058−1063 * |
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| CN105906566A (en) | 2016-08-31 |
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