CN106883245A - A kind of benzisoxa furfuran compound with removing free radical effect and preparation method and application - Google Patents
A kind of benzisoxa furfuran compound with removing free radical effect and preparation method and application Download PDFInfo
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- CN106883245A CN106883245A CN201710214686.8A CN201710214686A CN106883245A CN 106883245 A CN106883245 A CN 106883245A CN 201710214686 A CN201710214686 A CN 201710214686A CN 106883245 A CN106883245 A CN 106883245A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/04—Ortho-condensed systems
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
- A24B15/241—Extraction of specific substances
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/28—Treatment of tobacco products or tobacco substitutes by chemical substances
- A24B15/30—Treatment of tobacco products or tobacco substitutes by chemical substances by organic substances
- A24B15/36—Treatment of tobacco products or tobacco substitutes by chemical substances by organic substances containing a heterocyclic ring
- A24B15/40—Treatment of tobacco products or tobacco substitutes by chemical substances by organic substances containing a heterocyclic ring having only oxygen or sulfur as hetero atoms
- A24B15/403—Treatment of tobacco products or tobacco substitutes by chemical substances by organic substances containing a heterocyclic ring having only oxygen or sulfur as hetero atoms having only oxygen as hetero atoms
- A24B15/406—Treatment of tobacco products or tobacco substitutes by chemical substances by organic substances containing a heterocyclic ring having only oxygen or sulfur as hetero atoms having only oxygen as hetero atoms in a five-membered ring
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Abstract
The present invention relates to a kind of with benzisoxa furfuran compound for removing free radical effect and preparation method and application, belong to technical field of phytochemistry.The benzisoxa furfuran compound is isolated from tobacco, and Compound nomenclature is：The dimethyl 2 of 6,8 dihydro 2,2HFurans [3,4g] chromene, it is English entitled：6,8‑dihydro‑2,2‑dimethyl‑2H‑furo[3,4‑g] chromene, its molecular formula is C13H14O2, structure such as formula（I）It is shown：, formula（I）.Its preparation method is, with tobacco as raw material, to be obtained through medicinal extract extraction, organic solvent extraction, MCI decolourings, silica gel column chromatography and high performance liquid chromatography separation step.Antioxidation activity and free-radical scavenging activity screening are carried out to the compound, experimental result shows that the compound shows stronger anti-oxidant and free-radical scavenging activity.
Description
Technical field
The invention belongs to technical field of phytochemistry, and in particular to a kind of to extract the benzisoxa furan for obtaining first from tobacco
Class of muttering compound and its extracting method.Meanwhile, the compounds of this invention has preferable antioxidation activity, and application can have in cigarette
The removing Free Radicals In Cigarette Smoke of effect.
Background technology
Tobacco drafts a document that Solanaceae tobacco is annual or limited herbaceos perennial, wherein being cultivated with tobacco N.tabacum L.
Most wide, its ripe blade is the important source material of cigarette industry, is a kind of economic worth crop higher.Tobacco removes and is mainly used in
Suck outer, also other relatively broad purposes.Meanwhile, tobacco is the plant of chemical composition complexity the most in the world, secondary generation
Thank to product to enrich very much, by the research of decades, people identify the monomer chemistries material for coming from tobacco and just exceed at present
Kind more than 3000, and also many compositions not yet identify come.
Research shows, outside the routine chemical componentses such as isolating protein, amino acid, sugar, phosphorus, calcium, potassium, also containing a large amount of in tobacco
Secondary metabolite, type of compounds includes flavones, cumarin, tannin, furans, benzisoxa furans, Phenylpropanoid Glycosides, terpene, biology
Alkali etc..These compounds have multiple biological activities, in antibacterial, anti-inflammatory, anti-mutation, step-down, clearing heat and detoxicating, calm, diuresis, anti-
The many aspects such as oxidation, anticancer, anti-cancer, suppression lipase have remarkable result.Furfuran compound belongs to heterocyclic, natural
Also there is the document report for being widely present such compound in plant.In addition to extensive pharmacological action, furans or a class weight
The volatile compound wanted, is naturally occurring in various fruit and spice berry, such compound be widely used in various beverages,
In the formula of the flavoring essences such as bakery product.
A kind of present invention isolated benzisoxa furfuran compound from tobacco, activity research shows that the compound has
There is preferable antioxidation activity, be especially used as tobacco additive agent, with removing free radical antioxidation activity well
Effect, has positive effect to improving cigarette quality.The removing cigarette cigarette of benzisoxa furfuran compound is found in current tobacco
Gas free radical effect yet there are no pertinent literature and report.
The content of the invention
The first object of the present invention is to provide a kind of benzisoxa furfuran compound；Second purpose is to provide the benzene
And the preparation method of different furfuran compound；3rd purpose is to provide the application of the benzisoxa furfuran compound, mainly
For removing the free radical in cigarette mainstream flue gas.
To achieve the above object, the technical solution adopted by the present invention is as follows：
The first object of the present invention is achieved in that a kind of with the benzisoxa furans chemical combination for removing free radical effect
Thing is isolated from tobacco, and its molecular formula is C13H14O2, shown in structure such as formula (I)：
The compound is light yellow gum thing, is named as：6,8- dihydro -2,2- dimethyl -2H- furans [3,4-g] chromene,
English is entitled：6,8-dihydro-2,2-dimethyl-2H-furo[3,4-g]chromene.
The second object of the present invention is achieved in that above-mentioned with the benzisoxa furans chemical combination for removing free radical effect
The preparation method of thing, be with tobacco as raw material, successively through medicinal extract extract, organic solvent extraction, MCI decolourize, silica gel column chromatography and
High performance liquid chromatography separation step is obtained, specially：
A, medicinal extract are extracted：Tobacco is crushed to 20~40 mesh, 2~5 are extracted with 5~10 times of solvent supersonics of tobacco weight
It is secondary, 30~60 minutes every time, merge extract solution and filter, filtrate decompression is concentrated into the 1/4 of original volume, stands, and filters sediment,
Gained filtrate decompression is condensed into medicinal extract a afterwards；
B, organic solvent extraction：To the water that weight is 1~2 times of medicinal extract a weight is added in medicinal extract a, then with the body such as water
Long-pending organic solvent is extracted 3~5 times, merges organic solvent extraction phase, and the organic solvent extraction phase decompression for obtaining will be merged afterwards
It is condensed into medicinal extract b；
C, MCI decolourize：It is that 50~100% methyl alcohol are water-soluble to the volumetric concentration that addition in medicinal extract b is 3~5 times of medicinal extract b weight
Liquid, after medicinal extract b is completely dissolved, upper MCI posts are that 90%-95% methanol aqueous solutions are eluted with volumetric concentration, merge wash-out
Liquid, is concentrated under reduced pressure into medicinal extract c by the eluent after merging afterwards；
D, silica gel column chromatography：By silica gel column chromatography on medicinal extract c, dress post silica gel is 160~200 mesh, and consumption is medicinal extract c weight
6~10 times of amounts；It is 1 with volume ratio:0~0:1 chloroform and acetone mixed organic solvents gradient elution, collects the gradient of each gradient
Eluent is simultaneously concentrated under reduced pressure, and is monitored through TLC, merges identical part；
E, high performance liquid chromatography separation：It is 7 by volume ratio:The eluent that 3 chloroform-acetone is afforded, through efficient liquid
Phase chromatographic separation and purification, obtains final product described benzisoxa furfuran compound.
It is further preferred that the solvent of the step A is aqueous acetone solution, volume that volumetric concentration is 70~100%
The methanol aqueous solution that concentration is 90~100% ethanol water or volumetric concentration is 90~100%.
It is further preferred that the organic solvent of the step B is dichloromethane, chloroform, ethyl acetate, ether or stone
Oily ether.
It is further preferred that medicinal extract c is first medicinal extract c 1.5 with weight before through silica gel column chromatography in the D steps
~3 times of acetone or methyl alcohol dissolving, is then 80~100 mesh silica gel mixed samples of 0.8~1.2 times of medicinal extract c with weight, afterwards
Loading.
It is further preferred that in the D steps, during gradient elution, the chloroform and acetone mixed organic solvents for being used
Volume ratio be followed successively by 20:1、9:1、8:2、7:3、6:4 and 1:1, each gradient elution to TLC point plates without point after (the i.e. gradient
After can not eluting material), change next gradient elution.
It is further preferred that the high performance liquid chromatography separation purifying of the E steps is with first that volumetric concentration is 40%
Alcohol solution is mobile phase, 15~25ml/min of flow velocity, with 21.2 × 250mm, 5 μm of the Zorba x PrepHT anti-phase systems of GF
Standby post is fixing phase, and UV-detector Detection wavelength is 278nm, the μ L of each sample introduction 10~100, collects the chromatographic peak of 41.6min,
It is evaporated after repeatedly adding up.
The structure of the benzisoxa furfuran compound that method described above is prepared is measured by the following method：
The compounds of this invention is light yellow gum thing；
HRESI-MS shows that its quasi-molecular ion peak is 225.0898 [M+Na]+(calculated value 225.0891), with reference to1H NMR
Determine that its molecular formula is C with DEPT spectrums13H14O2, degree of unsaturation is 7.
Aromatic ring (1610,1561 and 1472cm are shown in infrared spectrum-1) resonance absorbing peak.And ultraviolet spectra 210,
240th, 278nm has absorption maximum to there may be aromatic ring structure in also illustrate that compound.
Compound1H and13C H NMR spectroscopies (such as table 1, Fig. 1 and Fig. 2) show that it contains 13 carbon and 14 hydrogen, including 1
1,2,4,5- quaternary phenyl ring (C-4~C-7, C-4a and C-7a, H-7), 1 humorous dimethyl chromene ring (C-1'~C-5', H-
1', H-2' and H6- 4', 5'), two methylene (C-1 and C-3 of oxidation；H2- 1 and H2-3).Further analyze its HMBC related
Signal, according to H2- 1 and C-3, C-4a, C-7, C-7a；H2- 3 and C-1, C-4, C-4a, C-7a；H-4 and C-3, C-4a, C-7a；With
And H-7 (such as Fig. 3) related to the HMBC of C-1, C-4a, C-7a can speculate that compound is benzisoxa furans class formation.
H-1' and C-4, C-5, C-6 can be observed in the HMBC spectrums of compound；H-2' and C-5, and H-4's and C-1' is remote
Cheng Xiangguan, can speculate that humorous dimethyl chromene ring is substituted in C-5 and C-6 of compound, and C-1' carbon is connected to phenyl ring
C-4.
So far, the structure of compound is determined, and is named as：6,8- dihydros -2,2- dimethyl -2H- furans [3,4-g]
Chromene.
Infrared, the ultraviolet and mass spectrometric data of compound：UV (methyl alcohol), λmax(logε)278(3.82)、240(3.68)、210
(4.05)nm；IR (pressing potassium bromide troche)：νmax 3080、2918、1610、1561、1472、1328、1275、1147、1051、
864cm-1；1H NMR and13C NMR datas (500 and 125MHz, CDCl3), it is shown in Table 1；[the M+ of positive ion mode ESIMS m/z 225
Na]+；Positive ion mode HRESIMS m/z 225.0898 [M+Na]+(calculated value C13H14NaO2, 225.0891).
The compounds of this invention of table 1.1H NMR and13C NMR datas (CDCl3)
What the third object of the present invention was realized in：
Benzisoxa furfuran compound of the present invention is used as the application for preparing antioxidant.
Antioxidation activity test is carried out to the compounds of this invention, antioxidation activity is with the big of scavenging ability of DPPH free radical
Small expression；Ethanol solution with 50 μ g/mL is primary dcreening operation concentration, determines its activity for removing lipid free radical DPPH.Take one piece
The orifice plates of costar 96, (concentration is 6.5 × 10 to the DPPH ethanol solutions of addition Fresh5Mol/L) 190 μ L/ holes, add this
Invention compound sample l0 μ L/ holes, blank well adds l0 μ L physiological saline, fully mixes, with lucifuge at room temperature after shrouding film shrouding
30 minutes are stood, in each hole absorbance is determined on analyzer on UV2401 spectrophotometers, measure wavelength is 517nm；Sample
Lipid free radical DPPH clearance rates are calculated as follows:
DPPH clearance rates (%)=(ABlank-ASample)/ABlank× 100%
ABlank：Blank control group absorbance；ASample：Plus sample sets absorbance.
Parallel 5 detections of sample, it is 4.36 μ g/L to calculate median elimination concentration IC50 measurement results, shows chemical combination of the present invention
Thing has good antioxidation activity and free-radical scavenging activity.
Application of the benzisoxa furfuran compound of the present invention in the free radical in removing cigarette smoke is provided simultaneously.
The test of Free Radicals In Cigarette Smoke effect to the compounds of this invention remove：
The Ye Zuwei of cigarette：Upper tobacco leaf is 15%, and middle part tobacco leaf is 48%, and lower tobacco leaf is 23%, expanded cut stem
8%, reconstituted tobacoo 6%；Using acetate fiber mouth rod, cigarette plug paper air permeability is 4500CU；Cigarette paper grammes per square metre is 50g/m2、
Air permeability is 80CU, and air permeability of tipping paper is 200CU.The weight of finished cigarettes cigarette is 0.93 ± 0.02g, and circumference is
24.5mm, length is 84mm (wherein mouth rod length is 25mm).
Test compound (benzisoxa furfuran compound of the present invention) is uniformly added to cigarette filter with essence injector
In tow, every cigarette addition is 0.5~5.0mg, and is control to be not added with the cigarette of test compound.
The cigarette duct automatic smoking machine smoking at the standard conditions of RM200 types 20, total particulate matter in mainstream smoke 44mm swords
Bridge filter disc is trapped, and gas phase portion is trapped with sampling pipe；The benzene of grain phase free radical 0.05mol/L N- tertiary butyls-α-phenyl nitrogen cave
Solution be extractant from cambridge filter dissolution, and wash cambridge filter, constant volume obtains a phase free radical test fluid.Gaseous phase free radical is used
Free radical sampling pipe, gathers, after cigarette smoking is complete by absorbent of the benzole soln of 0.05mol/L N- tertiary butyls-α-phenyl nitrogen cave
Gaseous phase free radical sampling pipe is taken out, while rinsing ventilation inner tube outside and sampling inside pipe wall for 3 times with a small amount of absorbent point, is merged and is inhaled
Liquid and cleaning solution are received, gaseous phase free radical sample liquid is obtained.
Free radical, ESR analysis experiment conditions are determined with EPR spectrometer：Central magnetic field=3.385T, sweeps width=0.500T,
Microwave frequency=1.5GHz, sweep time=2min, scanning times=5, multiplication factor=103~105(adjusted according to peak height
It is whole), the μ L of amount of samples 20；According to the change of calculated by peak area gas phase in ESR collection of illustrative plates and grain phase free radical quantity.
Test result indicate that：5 experiments are carried out, is compared with control sample, add the cigarette gas phase freedom of the compounds of this invention
Base reduced rate is 16~24%, and between 18~25%, the compound is in cigarette mainstream flue gas for grain phase free radical reduced rate
Free radical has definite elimination effect.
Compared with prior art, its advantage is the present invention：
Benzisoxa furfuran compound of the present invention is separated from tobacco first, by nuclear magnetic resonance and mass spectrum
It is benzisoxa furfuran compound that assay method is determined, and characterizes its concrete structure.Experiment proves the compounds of this invention tool
There is good antioxidation activity and free-radical scavenging activity.
Brief description of the drawings
Fig. 1 be benzisoxa furfuran compound of the present invention carbon-13 nmr spectra (13C NMR)；
Fig. 2 for benzisoxa furfuran compound of the present invention proton nmr spectra (1H NMR)；
Fig. 3 is the related figures of crucial HMBC of benzisoxa furfuran compound of the present invention.
Specific embodiment
With reference to embodiment, the present invention is described in further detail.
It will be understood to those of skill in the art that the following example is merely to illustrate the present invention, and should not be regarded as limiting this hair
Bright scope.In the examples where no specific technique or condition is specified, according to the technology or condition described by document in the art
Or carried out according to product description.Agents useful for same or the unreceipted production firm person of instrument, being can be by buying what is obtained
Conventional products.
Benzisoxa furfuran compound of the present invention, is isolated from tobacco, and its molecular formula is C13H14O2, tool
There are following structures：
Compound nomenclature is：6,8- dihydro -2,2- dimethyl -2H- furans [3,4-g] chromene, English is entitled：6,8-
dihydro-2,2-dimethyl-2H-furo[3,4-g]chromene。
The preparation method of benzisoxa furfuran compound of the present invention, is with tobacco material, through medicinal extract extraction, You Jirong
Agent extraction, MCI are decolourized, silica gel column chromatography and high performance liquid chromatography separation step are obtained, specially：
A, medicinal extract are extracted：Tobacco is crushed to 20~40 mesh, 2~5 are extracted with 5~10 times of solvent supersonics of tobacco weight
It is secondary, 30~60 minutes every time, merge extract solution and filter, filtrate decompression is concentrated into the 1/4 of original volume, stands, and filters sediment,
Gained filtrate decompression is condensed into medicinal extract a afterwards；
B, organic solvent extraction：To the water that weight is 1~2 times of medicinal extract a weight is added in medicinal extract a, then with the body such as water
Long-pending organic solvent is extracted 3~5 times, merges organic solvent extraction phase, and the organic solvent extraction phase decompression for obtaining will be merged afterwards
It is condensed into medicinal extract b；
C, MCI decolourize：It is that 50~100% methyl alcohol are water-soluble to the volumetric concentration that addition in medicinal extract b is 3~5 times of medicinal extract b weight
Liquid, after medicinal extract b is completely dissolved, upper MCI posts are that 90%-95% methanol aqueous solutions are eluted with volumetric concentration, merge wash-out
Liquid, is concentrated under reduced pressure into medicinal extract c by the eluent after merging afterwards；
D, silica gel column chromatography：By silica gel column chromatography on medicinal extract c, dress post silica gel is 160~200 mesh, and consumption is medicinal extract c weight
6~10 times of amounts；It is 1 with volume ratio:0~0:1 chloroform and acetone mixed organic solvents gradient elution, collects the gradient of each gradient
Eluent is simultaneously concentrated under reduced pressure, and is monitored through TLC, merges identical part；
E, high performance liquid chromatography separation：It is 7 by volume ratio:The eluent that 3 chloroform-acetone is afforded, through efficient liquid
Phase chromatographic separation and purification, obtains final product described benzisoxa furfuran compound.
The solvent of the step A is aqueous acetone solution that volumetric concentration is 70~100%, volumetric concentration is 90~100%
Ethanol water or methanol aqueous solution that volumetric concentration is 90~100%.
The organic solvent of the step B is dichloromethane, chloroform, ethyl acetate, ether or petroleum ether.
Medicinal extract c is the acetone or first of 1.5~3 times of medicinal extract c with weight before through silica gel column chromatography, first in the D steps
Alcohol dissolves, and is then 80~100 mesh silica gel mixed samples of 0.8~1.2 times of medicinal extract c with weight, afterwards loading.
In the D steps, during gradient elution, the chloroform and the volume ratio of acetone mixed organic solvents for being used are followed successively by
20:1、9:1、8:2、7:3、6:4 and 1:1.
The high performance liquid chromatography separation purifying of the E steps is to flow with methanol aqueous solution that volumetric concentration is 40%
Phase, 15~25ml/min of flow velocity, with 21.2 × 250mm, 5 μm of Zorbax PrepHT GF reverse phase preparative columns are fixing phase, purple
External detector Detection wavelength is 278nm, the μ L of each sample introduction 10~100, collects the chromatographic peak of 41.6min, is evaporated after repeatedly adding up.
Tobacco of the present invention is not limited by area and kind, can realize the present invention.
Embodiment 1
Dry tobacco 4.4kg is taken, meal is broken to 30 mesh, carried with 10 times the 70% of tobacco weight of aqueous acetone solution ultrasound
Take 4 times, 60 minutes every time, extract solution merged；Extract solution is filtered, and is concentrated under reduced pressure into the 1/4 of original volume；Stand, filter sediment,
It is concentrated under reduced pressure into the medicinal extract a of 120g；
240g water is added in medicinal extract a, is extracted 5 times with the chloroform isometric with water, merge organic solvent extraction phase, decompression
It is condensed into 80g medicinal extract b；
The methyl alcohol dissolving of 240g is added in medicinal extract b, after medicinal extract b is completely dissolved, upper MCI posts are with 6 liters of volumetric concentrations
90% methanol aqueous solution is eluted, and merges eluent, is concentrated under reduced pressure to give 62g medicinal extract c；
The acetone solution of 93g is added in medicinal extract c, 100 mesh silica gel 62g is subsequently adding and is mixed sample, after mixing sample, with 200 mesh silicon
Glue 372g fills post；The chloroform and the volume ratio of acetone mixed organic solvents for being used are followed successively by 20:1、9:1、8:2、7:3、6:4 Hes
1:1, after each gradient elution to TLC point plates is without point, next gradient elution is changed, collect gradient eluent, concentrated under reduced pressure, warp
TLC is monitored, and merges identical part, obtains 6 part A-F, wherein, to the sample D (7 being collected into:3) part 12g, then with
40% methyl alcohol is mobile phase, and flow velocity 25ml/min, 21.2 × 250mm, 5 μm of Zorbax PrepHT GF reverse phase preparative columns are
Fixing phase, UV-detector Detection wavelength is 278nm, the μ L of each sample introduction 10, the chromatographic peak of 41.6min is collected, after repeatedly adding up
It is evaporated, obtains final product the benzisoxa furfuran compound.
Embodiment 2
Dry tobacco 10kg is taken, meal is broken to 40 mesh, with 5 times of methyl alcohol ultrasonic extractions of tobacco weight 5 times, every time 30
Minute, extract solution merges；Extract solution is filtered, and is concentrated under reduced pressure into the 1/4 of original volume；Stand, filter sediment, be concentrated under reduced pressure into
300g medicinal extract a；
350g water is added in medicinal extract a, is extracted 3 times with the dichloromethane isometric with water, merge organic solvent extraction phase,
It is concentrated under reduced pressure into 210g medicinal extract b；
The volumetric concentration of 630g is added in medicinal extract b for 50% methanol aqueous solution dissolves, after medicinal extract b is completely dissolved, on
MCI posts, with 10 liters of volumetric concentrations for 95% methanol aqueous solution is eluted, merge eluent, are concentrated under reduced pressure to give 150g medicinal extract c；
The acetone solution of 300g is added in medicinal extract c, 80 mesh silica gel 180g is subsequently adding and is mixed sample, with 180 mesh silica gel 1500kg
Dress post, mixes upper prop after sample；The chloroform and the volume ratio of acetone mixed organic solvents for being used are followed successively by 20:1、9:1、8:2、7:3、
6:4 and 1:1, after each gradient elution to TLC point plates is without point, next gradient elution is changed, collect gradient eluent, depressurize dense
Contracting, monitors through TLC, merges identical part, obtains 6 part A-F, wherein, to the sample D (7 being collected into:3) part 32g,
It is again mobile phase, flow velocity 15ml/min, 21.2 × 250mm, 5 μm of the Zorbax PrepHT G anti-phase systems of F with 40% methyl alcohol
Standby post is fixing phase, and UV-detector Detection wavelength is 278nm, the μ L of each sample introduction 80, collects the chromatographic peak of 41.6min, repeatedly
It is evaporated after cumulative, obtains final product the benzisoxa furfuran compound.
Embodiment 3
Dry tobacco 15kg is taken, meal is broken to 20 mesh, with 8 times the 90% of tobacco weight of methanol aqueous solution ultrasonic extraction
2 times, 45 minutes every time, extract solution merged；Extract solution is filtered, and is concentrated under reduced pressure into the 1/4 of original volume；Stand, filter sediment, subtract
Pressure is condensed into 460g medicinal extract a；
460g water is added in medicinal extract a, is extracted 4 times with the ethyl acetate isometric with water, merge organic solvent extraction phase,
It is concentrated under reduced pressure into 300g medicinal extract b；
The volumetric concentration of 1500g is added in medicinal extract b for 85% methanol aqueous solution dissolves, after medicinal extract b is completely dissolved, on
MCI posts, with 15 liters of volumetric concentrations for 93% methanol aqueous solution is eluted, merge eluent, are concentrated under reduced pressure to give 150g medicinal extract c；
The methyl alcohol dissolving of 450g is added in medicinal extract c, 100 mesh silica gel 120g is subsequently adding and is mixed sample, with 160 mesh silica gel 1.2kg
Dress post, mixes upper prop after sample；The chloroform and the volume ratio of acetone mixed organic solvents for being used are followed successively by 20:1、9:1、8:2、7:3、
6:4 and 1:1, after each gradient elution to TLC point plates is without point, next gradient elution is changed, collect gradient eluent, depressurize dense
Contracting, monitors through TLC, merges identical part, obtains 6 part A-F, wherein, to the sample D (7 being collected into:3) part 32g,
It is again mobile phase, flow velocity 20ml/min, 21.2 × 250mm, 5 μm of the Zorbax PrepHT G anti-phase systems of F with 40% methyl alcohol
Standby post is fixing phase, and UV-detector Detection wavelength is 278nm, the μ L of each sample introduction 100, collects the chromatographic peak of 41.6min, repeatedly
It is evaporated after cumulative, obtains final product the benzisoxa furfuran compound.
Embodiment 4
Embodiment 4 is with the difference of embodiment 3, and during ultrasonic extraction, solvent for use is 95% ethanol water.Extraction
When, the organic solvent for using is ether.
Embodiment 5
Embodiment 5 is with the difference of embodiment 3, and during ultrasonic extraction, solvent for use is 90% aqueous acetone solution.Extraction
When, the organic solvent for using is petroleum ether.
Embodiment 6
The structure of the benzisoxa furfuran compound that embodiment 1 is prepared is measured by the following method；The present invention
Compound is light yellow gum thing；HRESI-MS shows that its quasi-molecular ion peak is 225.0898 [M+Na]+(calculated value
225.0891), with reference to1H NMR and DEPT spectrum determine that its molecular formula is C13H14O2, degree of unsaturation is 7.Shown in infrared spectrum
Aromatic ring (1610,1561 and 1472cm-1) resonance absorbing peak.And ultraviolet spectra has absorption maximum also to say in 210,240,278nm
Aromatic ring structure is there may be in clear compound.Compound1H and13C H NMR spectroscopies (table 1, Fig. 1 and Fig. 2) show that it contains 13
Carbon and 14 hydrogen, including 11,2,4,5- quaternary phenyl ring (C-4~C-7, C-4a and C-7a, H-7), 1 humorous diformazan primary colours
Alkene ring (C-1'~C-5', H-1', H-2' and H6- 4', 5'), two methylene (C-1 and C-3 of oxidation；H2- 1 and H2-3).Enter
One step analyzes its HMBC coherent signal, according to H2- 1 and C-3, C-4a, C-7, C-7a；H2- 3 and C-1, C-4, C-4a, C-7a；H-4
With C-3, C-4a, C-7a；And H-7 (Fig. 3) related to the HMBC of C-1, C-4a, C-7a can speculate that compound is benzisoxa furans
Class formation.H-1' and C-4, C-5, C-6 can be observed in the HMBC spectrums of compound；H-2' and C-5, and H-4's and C-1' is remote
Cheng Xiangguan, can speculate that humorous dimethyl chromene ring is substituted in C-5 and C-6 of compound, and C-1' carbon is connected to phenyl ring
C-4.So far, the structure of compound determined, and is named as Compound nomenclature and is：6,8- dihydro -2,2- dimethyl -2H-
Furans [3,4-g] chromene.
Embodiment 7
Compound prepared by Example 2-5, is light yellow gum thing.Determine same as Example 6, confirm to implement 2-5
The compound of preparation is the benzisoxa furfuran compound --- 6,8- dihydro -2,2- dimethyl -2H- furans [3,4-g] color
Alkene.
Embodiment 8
Antioxidation activity test is carried out to compound, antioxidation activity is with the size table of scavenging ability of DPPH free radical
Show；Ethanol solution with 50 μ g/mL is primary dcreening operation concentration, determines its activity for removing lipid free radical DPPH.Take one piece of costar
96 orifice plates, add the DPPH ethanol solutions (6.5 × 10 of Fresh5Mol/L) 190 μ L/ holes, add the compounds of this invention sample
L0 μ L/ holes, blank well adds l0 μ L physiological saline, fully mixes, and with after shrouding film shrouding, lucifuge stands 30 minutes at room temperature, in
Each hole absorbance is determined on UV2401 spectrophotometers on analyzer, measure wavelength is 517nm；Sample is to lipid free radical
DPPH clearance rates are calculated as follows:
DPPH clearance rates (%)=(ABlank-ASample)/ABlank× 100%
ABlank：Blank control group absorbance；ASample：Plus sample sets absorbance.
Parallel 5 detections of sample, it is 4.36 μ g/L to calculate median elimination concentration IC50 measurement results, shows that compound has
Good antioxidation activity and free-radical scavenging activity.
Embodiment 9
The removing Free Radicals In Cigarette Smoke measure of merit of compound：
(1) Ye Zuwei of cigarette：Upper tobacco leaf is 15%, and middle part tobacco leaf is 48%, and lower tobacco leaf is 23%, expanded cut stem
8%, reconstituted tobacoo 6%；Using acetate fiber mouth rod, cigarette plug paper air permeability is 4500CU；Cigarette paper grammes per square metre is 50g/m2、
Air permeability is 80CU, and air permeability of tipping paper is 200CU.The weight of finished cigarettes cigarette is 0.93 ± 0.02g, and circumference is
24.5mm, length is 84mm (wherein mouth rod length is 25mm).
Test compound essence injector is uniformly added in cigarette-filter tow, and every cigarette addition is 0.5
~5.0mg, and be control to be not added with the cigarette of test compound.
(2) the cigarette duct automatic smoking machine smoking at the standard conditions of RM200 types 20, total particulate matter in mainstream smoke 44mm
Cambridge filter is trapped, and gas phase portion is trapped with sampling pipe；The grain phase free radical tertiary butyls of N- containing 0.05mol/L-α-phenyl nitrogen cave
Benzole soln be extractant from cambridge filter dissolution, and wash cambridge filter, constant volume obtains a phase free radical test fluid.Gas phase freedom
Base free radical sampling pipe, gathers, cigarette smoking by absorbent of the benzole soln of 0.05mol/L N- tertiary butyls-α-phenyl nitrogen cave
Gaseous phase free radical sampling pipe is taken out after complete, while rinsing ventilation inner tube outside and sampling inside pipe wall for 3 times with a small amount of absorbent point, is closed
And absorbing liquid and cleaning solution, obtain gaseous phase free radical sample liquid.
(3) free radical, ESR analysis experiment conditions are determined with EPR spectrometer：Central magnetic field=3.385T, sweep it is wide=
0.500T, microwave frequency=1.5GHz, sweep time=2min, scanning times=5, multiplication factor=103~105(according to peak height
It is adjusted), the μ L of amount of samples 20；According to the change of calculated by peak area gas phase in ESR collection of illustrative plates and grain phase free radical quantity.
Test result indicate that：5 experiments are carried out, is compared with control sample, add the cigarette gas phase freedom of the compounds of this invention
Base reduced rate is 16~24%, and between 18~25%, the compound is in cigarette mainstream flue gas for grain phase free radical reduced rate
Free radical has definite elimination effect.
General principle of the invention, principal character and advantages of the present invention has been shown and described above.The technology of the industry
Personnel it should be appreciated that the present invention is not limited to the above embodiments, simply explanation described in above-described embodiment and specification this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and modifications of the present invention are possible, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appending claims and its
Equivalent thereof.
Claims (9)
1. it is a kind of with the benzisoxa furfuran compound for removing free radical effect, it is characterised in that the benzisoxa furans
Compound is isolated from tobacco, is named as：6,8- dihydro -2,2- dimethyl -2H- furans [3,4-g] chromene, English name
For：6,8-dihydro-2,2-dimethyl- 2H-furo[3,4-g] chromene, its molecular formula is C13H14O2, structure such as formula
（I）It is shown：
, formula（I）.
2. described in a kind of claim 1 with remove free radical effect benzisoxa furfuran compound preparation method, its
It is characterised by, with tobacco as raw material, successively through medicinal extract extraction, organic solvent extraction, MCI decolourings, silica gel column chromatography and efficient liquid
Phase chromatrographic separation step is obtained, specially：
A, medicinal extract are extracted：Tobacco is crushed to 20 ~ 40 mesh, 2 ~ 5 times are extracted with the solvent supersonics of 5 ~ 10 times of tobacco weight, every time 30
~ 60 minutes, merge extract solution and filter, filtrate decompression is concentrated into the 1/4 of original volume, stands, and sediment is filtered, afterwards by gained
Filtrate decompression is condensed into medicinal extract a；
B, organic solvent extraction：To the water that weight is 1 ~ 2 times of medicinal extract a weight is added in medicinal extract a, then have with isometric with water
Machine solvent extraction 3 ~ 5 times, merges organic solvent extraction phase, the organic solvent extraction phase for obtaining will be merged afterwards and is concentrated under reduced pressure into leaching
Cream b；
C, MCI decolourize：It is 50 ~ 100% methanol aqueous solutions to the volumetric concentration that addition in medicinal extract b is 3 ~ 5 times of medicinal extract b weight, waits to soak
After cream b is completely dissolved, upper MCI posts are that 90%-95% methanol aqueous solutions are eluted with volumetric concentration, merge eluent, afterwards will
Eluent after merging is concentrated under reduced pressure into medicinal extract c；
D, silica gel column chromatography：By silica gel column chromatography on medicinal extract c, dress post silica gel is 160 ~ 200 mesh, and consumption is medicinal extract c weight 6 ~ 10
Measure again；It is 1 with volume ratio:0~0:1 chloroform and acetone mixed organic solvents gradient elution, collects the gradient eluent of each gradient
And be concentrated under reduced pressure, monitored through TLC, merge identical part；
E, high performance liquid chromatography separation：It is 7 by volume ratio:The eluent that 3 chloroform-acetone is afforded, through high-efficient liquid phase color
Spectrum is isolated and purified, and obtains final product described benzisoxa furfuran compound.
3. it is according to claim 2 with remove free radical effect benzisoxa furfuran compound preparation method, its
It is characterised by, the solvent of the step A is aqueous acetone solution, second that volumetric concentration be 90 ~ 100% of the volumetric concentration for 70 ~ 100%
Alcohol solution or the methanol aqueous solution that volumetric concentration is 90 ~ 100%.
4. it is according to claim 2 with remove free radical effect benzisoxa furfuran compound preparation method, its
It is characterised by, the organic solvent of the step B is dichloromethane, chloroform, ethyl acetate, ether or petroleum ether.
5. it is according to claim 2 with remove free radical effect benzisoxa furfuran compound preparation method, its
It is characterised by, medicinal extract c is the acetone or first of 1.5 ~ 3 times of medicinal extract c with weight before through silica gel column chromatography, first in the D steps
Alcohol dissolves, and is then 80 ~ 100 mesh silica gel mixed samples of 0.8 ~ 1.2 times of medicinal extract c with weight, afterwards loading.
6. it is according to claim 2 with remove free radical effect benzisoxa furfuran compound preparation method, its
It is characterised by, in the D steps, during gradient elution, the chloroform and the volume ratio of acetone mixed organic solvents for being used are followed successively by
20:1、9:1、8:2、7:3、6:4 and 1:1, after each gradient elution to TLC point plates is without point, change next gradient elution.
7. it is according to claim 2 with remove free radical effect benzisoxa furfuran compound preparation method, its
The high performance liquid chromatography separation purifying for being characterised by the E steps be with methanol aqueous solution that volumetric concentration is 40% as mobile phase,
15 ~ 25ml/min of flow velocity, with 21.2 × 250mm, 5 μm of Zorbax PrepHT GF reverse phase preparative columns are fixing phase, ultraviolet inspection
It is 278 nm, the μ L of each sample introduction 10 ~ 100 to survey device Detection wavelength, collects the chromatographic peak of 41.6 min, is evaporated after repeatedly adding up.
8. described in claim 1 with removing the benzisoxa furfuran compound of free radical effect as preparing antioxidant
Using.
9. the benzisoxa furfuran compound with removing free radical effect described in claim 1 is in cigarette smoke is removed
Application in free radical.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108409545A (en) * | 2018-03-27 | 2018-08-17 | 云南中烟工业有限责任公司 | A kind of ether compound, preparation method and application |
| CN111567854A (en) * | 2020-05-12 | 2020-08-25 | 云南中烟工业有限责任公司 | Benzisothiafuranolactone compound, preparation method thereof, antioxidant and additive containing benzisothiafuranolactone compound and application of benzisothiafuranolactone compound |
| CN114920723A (en) * | 2022-03-14 | 2022-08-19 | 云南中烟工业有限责任公司 | Chromene compound with antioxidant activity and preparation method and application thereof |
| CN115385927A (en) * | 2022-06-10 | 2022-11-25 | 中国科学院西北高原生物研究所 | Separation process and application of furocoumarin natural free radical scavenger in dracocephalum heterophyllum benth |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010037244A2 (en) * | 2008-10-01 | 2010-04-08 | Givaudan Sa | Organic compounds |
| CN106117171A (en) * | 2016-06-29 | 2016-11-16 | 云南中烟工业有限责任公司 | A kind of supercritical fluid chromatography prepares the benzisoxa furfuran compound methods and applications in Nicotiana tabacum L. with antibacterial activity |
| CN106380473A (en) * | 2016-08-29 | 2017-02-08 | 云南中烟工业有限责任公司 | Novel lactone compound, preparation method thereof and application thereof |
-
2017
- 2017-04-01 CN CN201710214686.8A patent/CN106883245B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010037244A2 (en) * | 2008-10-01 | 2010-04-08 | Givaudan Sa | Organic compounds |
| CN106117171A (en) * | 2016-06-29 | 2016-11-16 | 云南中烟工业有限责任公司 | A kind of supercritical fluid chromatography prepares the benzisoxa furfuran compound methods and applications in Nicotiana tabacum L. with antibacterial activity |
| CN106380473A (en) * | 2016-08-29 | 2017-02-08 | 云南中烟工业有限责任公司 | Novel lactone compound, preparation method thereof and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| RAGHAO S. MALI，等: "Naturally Occurring Prenylated Phthalides: First Total Synthesis of Salfredin B11", 《JOURNAL OF CHEMICAL RESEARCH, SYNOPSES》 * |
Cited By (6)
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|---|---|---|---|---|
| CN108409545A (en) * | 2018-03-27 | 2018-08-17 | 云南中烟工业有限责任公司 | A kind of ether compound, preparation method and application |
| CN108409545B (en) * | 2018-03-27 | 2021-01-29 | 云南中烟工业有限责任公司 | Ether compound, preparation method and application thereof |
| CN111567854A (en) * | 2020-05-12 | 2020-08-25 | 云南中烟工业有限责任公司 | Benzisothiafuranolactone compound, preparation method thereof, antioxidant and additive containing benzisothiafuranolactone compound and application of benzisothiafuranolactone compound |
| CN111567854B (en) * | 2020-05-12 | 2022-02-08 | 云南中烟工业有限责任公司 | Benzisothiafuranolactone compound, preparation method thereof, antioxidant and additive containing benzisothiafuranolactone compound and application of benzisothiafuranolactone compound |
| CN114920723A (en) * | 2022-03-14 | 2022-08-19 | 云南中烟工业有限责任公司 | Chromene compound with antioxidant activity and preparation method and application thereof |
| CN115385927A (en) * | 2022-06-10 | 2022-11-25 | 中国科学院西北高原生物研究所 | Separation process and application of furocoumarin natural free radical scavenger in dracocephalum heterophyllum benth |
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