CN105985390A - Standard extract in Inula salsoloides and analytical method and application thereof - Google Patents
Standard extract in Inula salsoloides and analytical method and application thereof Download PDFInfo
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Abstract
The invention discloses standard extracts in Inula salsoloides and an analytical method and application thereof. 16 compounds are isolated from Inula salsoloides, structures of the compounds are identified by the comprehensive use of modern spectroscopic techniques such as nuclear magnetic resonance spectroscopy, ultraviolet spectroscopy and mass spectroscopy, a new compound formula is C21H32O8, crystal color is white, and the new compound is named as II type eupatolide; activity studies discover that the compound IS-10 can repel the pests such as Phyllotreta striolata; the compounds IS-3 and IS-16 have antifeedant function, and it is proved that sesquiterpene compounds in Inula salsoloides are ingredients effective in providing pesticidal activity. The invention also discloses preparation and quantitative analytical method for standard extracts of Inula salsoloides, and activity testing discovers that the extracts have significant avoiding and antifeedant functions for the pests such as Phyllotreta striolata.
Description
Technical field
The present invention relates to a kind of expelling parasite desinsection chemical active ingredient extracting from herbal medicine and preparation thereof and analysis method, tool
Body, relates to the separation of Pesticidal compound and a noval chemical compound in salsola-like inula flower and herb and activity standardizes expelling parasite insect-killing extract with it
Preparation and quantitative analysis method.
Background technology
Husky multiple japanese bearbind is flower and the herb of composite family Inula plant sand ground inular flower, calls bald woman lama careless, yellow, Huang
Hua Song, knotweed Piao.It is distributed widely in Central China and west area, in illiteracy, Ji, Shanxi, Shan, new, North Qinghai and east and the Liao Dynasty
There is growth in peaceful western part.It is abroad distributed in Central Asia of the Mongolian and former Soviet Union.General on grassland, Gobi deserf, fixing sand dune
And loess plateau can grow, also see growth in alluvial tract, riverbank, less demanding for height above sea level, height above sea level 500 to 2000 meters
There is distribution, at moist sandy soil, the growth in flakes on the ground of moving dunes edge and bank sand.It is a kind of adaptable Asia
Shrub plant.Whole plant for medical use, Xia Qiu gathers, and dries standby.Taste is pungent cool in nature, has the function of heat-clearing, diuresis.
Existing document report, salsola-like inula flower and herb extract has certain antimicrobial and insecticidal activity.Qu Qiuyun etc. study discovery, husky
Inular flower embodies very strong biologically active in terms of expelling parasite desinsection.It is to big wrinkle gill cockchafer, anomala mongolica, eight word white-browed skies
Three kinds of insects such as moth all demonstrate higher toxic effect, and three kinds of insect death rates are more than 90%.Zhang Hongli etc. study discovery,
Food refusal and toxic effect to mythimna separata for the salsola-like inula flower and herb extract is all higher than 80%, shows that it has preferable food refusal and poisoning effect to mythimna separata
Really.Salsola-like inula flower and herb in Yanchi county Ningxia by as farmland weed process.
Lin Kun etc. study the biologically active to diamondback moth for the salsola-like inula flower and herb ethanol extract, and in research process, author is in indoor survey
Determined salsola-like inula flower and herb ethanol extract to 3 age the tagging of diamondback moth larvae, food refusal, repellent rate, stomach toxicity, Developing restraint grow 5
Plant biological action.Result shows: salsola-like inula flower and herb ethanol extract has tagging and food refusal clearly to diamondback moth 3 instar larvae
Activity, corrected mortality and anti-food rate all increase with the rising of extract concentrations；Ethanol extract diamondback moth is also had stomach toxicity,
Developing restraint grows isoreactivity.Salsola-like inula flower and herb different parts ethanol extract also with diamondback moth for examination worm, is killed by Zhao Tang etc.
Worm expression activitiy, filters out activity highest portion position, then uses different solvents to carry out extraction point to it, obtains different solvents extraction
Each extract tagging and antifeedant activity to diamondback moth is tested after thing.Result shows, salsola-like inula flower and herb alcohol extract chloroform extract
Best to the activity of diamondback moth, and herb is compared to other each positions (flower, leaf, root, stem etc.) tagging and food refusal to diamondback moth
Activity is the highest.Inferring that this should be due to synergy produced by each position of salsola-like inula flower and herb, the desinsection i.e. containing in herb is lived
The all high reason in the property arbitrary position of composition, each extractive part creates synergistic effect together.Salsola-like inula flower and herb is to diamondback moth
Antifeedant activity is with processing the growth of time, and effect gradually weakens, it may be possible to have the composition of antifeedant activity easily to wave in salsola-like inula flower and herb
Send out or produce drug resistance in microzooid and cause active duration short.
When screening anticancer plants, Chen Jie etc. finds that leukaemia P-388 cell is had by the ether position in salsola-like inula flower and herb
There is stronger inhibitory action.1994, Bing-Nan Zhou et al. got two new sesquiterpene lactones from salsola-like inula flower and herb
15-deoxygenates sand ground britannin inulasalsolin and sand ground britannin inulasalsolide, and known to two
Compound eupatolide and budlein B, and find that 15-deoxygenates sand ground britannin (inulasalsolin) and sand ground
Britannin (inulasalsolide) demonstrates strong cell toxicological activity to KB and P-388 cell line.Domestic
The pharmacology activity research of the salsola-like inula flower and herb that person reports in recent years is fewer, and Li Yu equality report discovery salsola-like inula flower and herb has very strong
Bacteriostatic activity, has the inhibiting rate of 100% to multiple pathogens such as gibberella saubinetii, apple anthrax, Phytophthora capsici, the big spots of corn.
The Long-Time Service of traditional chemical agricultural chemicals brings all drawbacks, such as generation, the residual toxicity of harmful biomolecule resistant
And environmental pollution etc..The development of society, the needs of the public, promote people at the different aspect such as microbiology, Phytochemistry
Seek the novel pest control agent to human health and ecological environment security.This problem is based on abundant the checking winds and fixing drifting sand in northern Shensi
Natural plant resource salsola-like inula flower and herb, intends Primary Study and goes out its active site, for developing for efficient preventing and treating harmful organism
Plant insecticide lay theoretical foundation, the characteristic of the Modern Characteristic development in agricultural science and technology of China will be fully demonstrated.
Content of the invention
For solving the problems referred to above, the invention provides a kind of salsola-like inula flower and herb Plays extract and extracting method thereof and answer
With.
For achieving the above object, the technical scheme that the present invention takes is:
A kind of salsola-like inula flower and herb Plays extract, this extract includes following composition: Ahmedabad Rhein .B:2.122mg/g；
Ovum South America chrysanthemum element: 0.544mg/g；Sand ground britannin: 1.101mg/g；15-deoxygenates sand ground britannin: 0.855mg/
g；4 α, 5 beta epoxide ovum South America chrysanthemum element: 0.669mg/g；Eupatolide: 11.199mg/g；Apiolin: 0.362mg/g；Total amount is
16.852mg/g。
From salsola-like inula flower and herb, the chemical formula of one noval chemical compound of isolated is C21H32O8, crystal color is white, life
Entitled eupatolide-II, has 21 carbon atoms in structure, including 3 quaternary carbon atoms, 11 tertiary carbon atom CH, 5 secondary carbon
Atom CH2With 2 primary carbon CH3；Structure has a carbonyl signals δ 178.36 (C-12), four olefinic carbon atoms signal δ 139.21
(C-4) and δ 136.18 (C-1), δ 132.01 (C-5), δ 128.04 (C-10), an even oxygen anomeric carbon signals δ 103.13 (C-
, and δ 103.13,76.70,73.65,68.80,61.04,57.02 is one group of glucose signals 1 ')；5 secondary carbons, respectively
For δ 67.82 (C-13), 61.04 (C-6 '), 42.07 (C-9), 38.59 (C-3) and 25.24 (C-2)；11 tertiary carbon atoms remove
On sugar ring 6, are left as δ 132.01 (C-5), 128.04 (C-10), 57.02 (C-4 '), 48.54 (C-7) and 38.46 (C-
11)；Two quaternary carbons are δ 16.45 (C-15), 13.26 (C-14)；Anomeric carbon signals δ 103.13 (C-1 ') and hydrogen signal δ 4.12
(H-1 ') is joined directly together.
For solving the problems referred to above, present invention also offers carrying of a kind of salsola-like inula flower and herb Plays extract and noval chemical compound
Access method, comprises the steps:
S1, take dry salsola-like inula flower and herb herb 10kg, with 90% ethanol water room temperature cold soaking three of 6 times of volumes after pulverizing
Secondary, each 24h, filters, concentrated filtrate, obtains the dark brown paste of 0.93kg, and recycling design；
S2, by the paste of step S1 gained with suitable water-dispersible after, use dichloromethane successively, ethyl acetate extracts respectively
Taking three times, each extraction position is concentrated into medicinal extract more respectively, respectively obtains 308.8g dichloromethane extract, 81.62g acetic acid second
Ester extract, 297.0g aqueous fraction；
S3, the filter residue part continuation water-cooled by subsidy S1 gained extract and take, three times, and each 24h is concentrated into medicinal extract,
Salsola-like inula flower and herb water extract；
S4, the dichloromethane layer by step S2 gained, use 300g silica gel mixed sample, and rotation upper 1kg silicagel column after being evaporated, with oil
After ether-ethyl acetate gradient, rinse with methyl alcohol, according to TLC testing result in elution process, merge same section eluent
And concentrate, obtain 10g Fr.1,12g flow point Fr.3,18g flow point Fr.4,10g flow point Fr.5,8g flow point Fr.6, Fr.7；
S5, the flow point of above-mentioned gained is carried out lock out operation
S51, by the flow point Fr.2 of step S4 gained, through silica gel column chromatography, with petroleum ether-ethyl acetate gradient elution, will
It is divided into 4 sections of Fr2-1-Fr2-4, by Fr2-3 flow point, through silica gel column chromatography repeatedly, petroleum ether-ethyl acetate and petroleum ether-
Acetone system gradient elution, is recrystallizing repeatedly so as to obtain compound 36mg IS-7,54mg IS-8, by Fr2-4 through layer of silica gel repeatedly
Analysis obtains compound 15mg IS-9；
S52, by the flow point Fr.3 of step S4 gained, through silica gel column chromatography, petroleum ether-ethyl acetate gradient elution, wash-out
Process TLC tracing detection, merges same composition and obtains 3 sections of Fr3-1-Fr3-3, by Fr3-3 flow point through silica gel column chromatography repeatedly,
To compound 6mg IS-1,100mg IS-5,24mg IS-14；
S53, by the flow point Fr.4 of step S4 gained, through silica gel column chromatography, petroleum ether-ethyl acetate gradient elution, wash-out
Process TLC tracing detection, merges same composition and obtains 4 sections of Fr4-1-Fr4-4, by Fr4-2 flow point through silica gel column chromatography repeatedly,
To compound 18mg IS-2,10mg IS-4；By Fr4-3 flow point through silica gel column chromatography repeatedly, and combine MCI CHP-20P,
To compound 18mg IS-3,10mg IS-6,14mg IS-12；
S54, by the flow point Fr.5 of step S4 gained, through silica gel column chromatography, methylene chloride-methanol gradient elution, strip
Journey TLC tracing detection, merges same composition and obtains 5 sections of Fr5-1-Fr5-5, by Fr5-1 flow point through silica gel column chromatography repeatedly, and tie
Close Sephadex LH-20 and obtain compound 18mg IS-10,10mg IS-17；By Fr5-3 flow point through silica gel column chromatography repeatedly,
And combine MCI CHP-20P, obtain compound 45mg IS-11,10mg IS-13；By Fr5-4 flow point through silica gel column layer repeatedly
Analysis, purifies and obtains compound 7mg IS-15；
S55, by the flow point Fr.6 of step S4 gained, through silica gel column chromatography, methylene chloride-methanol gradient elution, strip
Journey TLC tracing detection, merges same composition and obtains 3 sections of Fr6-1-Fr6-3, by Fr6-3 flow point through silica gel column chromatography repeatedly, and tie
Close MCI CHP-20P, obtain compound 14mg IS-16.
Wherein, the silica gel in described step S4 is 100-200 mesh；In step S5, silica gel is 200-300 mesh.
Wherein, in described step S4, the gradient of petroleum ether-ethyl acetate is 200: 1-0: 1；Oil in described step S51
The gradient of ether-ethyl acetate is 100: 1-5: 1；In step S52, the gradient of petroleum ether-ethyl acetate is 60: 1-0: 1；Step S53
The gradient of middle petroleum ether-ethyl acetate is 40: 1-0: 1.
Wherein, in described step S55, the gradient of difluoromethane-methyl alcohol is 60: 1-1: 1
The method have the advantages that
From the Ethyl acetate extract of salsola-like inula flower and herb 90% ethanol extract, through silica gel, MCI CHP-20P, Sephadex
The various column chromatography chromatogram separation methods such as LH-20 hydroxypropyl sephadex, and combine TLC tracing detection and HPLC purity is divided
Analysis, 17 compounds of isolated from which, integrated use NMR spectrum (13C-NMR、1H-NMR、1H-1H COSY、
HMBC, HMQC, NOESY), the Modern spectroscopy such as ultraviolet spectra (UV) and mass spectrum (EI-MS) learn a skill, identify its structure.Including 1
Individual monoterpenes compound: cis-1-p-Menthene-3,6-diol (IS-1)；12 sesquiterpenoids (contain newization
Compound): Ahmedabad Rhein .B (IS-2), 1-Cyclodecene-2,5,6,9-tetrol, 1,8-dimethyl-7-(1-
Methylethyl)-, [(2 α, 5 β, 6 α, 7 α, 8 β, 9 α)] (IS-3), sand ground britannin, 4 α, 5 β-Epoxy-8 β-
Hydroxy-14-oxoacanthospermolide (IS-5), ovum South America chrysanthemum element (Ovatifolin, IS-9), 15-deoxygenates sand ground
Britannin (Inulasalsolin, IS-10), 4 α, 5 beta epoxide ovum South America chrysanthemum elements (4 α, 5 β-Epoxyovatifolin,
IS-11), sand ground britannin B (Inulasalsolide B, IS-12), eupatolide (Eupatolide, IS-13), 11
β, 13-dihydro eupatolide (11 β, 13-dihydro-Eupatolide, IS-14), II type eupatolide (eupatolide-
II, IS-15), 1 flavone compound: apiolin (Apigenin, IS-16) and 3 steroid compounds: cupreol (β-
Sitosterol, IS-6), stigmasterol (Stigmasterol, IS-7), Daucosterol (Daucosterol, IS-8).
Brief description
Fig. 1 is the molecular structure of a kind of salsola-like inula flower and herb Plays extract in the embodiment of the present invention.
Fig. 2 is the extraction flow chart of a kind of salsola-like inula flower and herb Plays extract in the embodiment of the present invention.
Fig. 3 is the high-efficient liquid phase chromatogram of a kind of salsola-like inula flower and herb Plays extract in the embodiment of the present invention.
Detailed description of the invention
In order to make objects and advantages of the present invention clearer, below in conjunction with embodiment, the present invention is carried out further
Describe in detail.It should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not used to limit this
Bright.
As it is shown in figure 1, embodiments provide a kind of salsola-like inula flower and herb Plays extract, this extract include as
Lower composition: Ahmedabad Rhein .B:2.122mg/g；Ovum South America chrysanthemum element: 0.544mg/g；Sand ground britannin: 1.101mg/g；15-
Deoxygenate sand ground britannin: 0.855mg/g；4 α, 5 beta epoxide ovum South America chrysanthemum element: 0.669mg/g；Eupatolide:
11.199mg/g；Apiolin: 0.362mg/g；Total amount is 16.852mg/g.One new chemical combination of isolated from salsola-like inula flower and herb
The chemical formula of thing is C21H32O8, crystal color is white, molecular weight 412.47.
Its H spectrum and C modal data are as shown in the table.
The H spectrum of table 1 salsola-like inula flower and herb Plays extract and C compose (DMSO-d6)
As in figure 2 it is shown, above-mentioned extract is prepared by the following method:
S1, take dry salsola-like inula flower and herb herb 10kg, with 90% ethanol water room temperature cold soaking three of 6 times of volumes after pulverizing
Secondary, each 24h, filters, concentrated filtrate, obtains the dark brown paste of 0.93kg, and recycling design；
S2, by the paste of step S1 gained with suitable water-dispersible after, use dichloromethane successively, ethyl acetate extracts respectively
Taking three times, each extraction position is concentrated into medicinal extract more respectively, respectively obtains 308.8g dichloromethane extract, 81.62g acetic acid second
Ester extract, 297.0g aqueous fraction；
S3, the filter residue part continuation water-cooled by subsidy S1 gained extract and take, three times, and each 24h is concentrated into medicinal extract,
Salsola-like inula flower and herb water extract；
S4, the dichloromethane layer by step S2 gained, mix sample with 300g silica gel (100-200 mesh), rotation upper 1kg silicon after being evaporated
Glue post, after petroleum ether-ethyl acetate (200: 1-0: 1) gradient elution, is rinsed by methyl alcohol, detects according to TLC in elution process
As a result, merge same section eluent and concentrate, obtaining 10g Fr.1,12g flow point Fr.3,18g flow point Fr.4,10g flow point
Fr.5,8g flow point Fr.6, Fr.7；
S5, by the flow point Fr.2 of step S4 gained, through silica gel (200-300 mesh) column chromatography, with petroleum ether-ethyl acetate
(100: 1-5: 1) gradient elution, is divided into 4 sections of Fr2-1-Fr2-4.Wherein Fr2-3 flow point, through silica gel column chromatography repeatedly
(200-300 mesh), petroleum ether-ethyl acetate and petroleum ether-acetone system gradient elution, is recrystallizing repeatedly so as to obtain compound
Fr2-4 (220mg) is obtained compound 15mg IS-9 through silica gel column chromatography repeatedly by 36mg IS-7,54mg IS-8；
By the flow point Fr.3 of step S4 gained, through silica gel (200-300 mesh) column chromatography, petroleum ether-ethyl acetate (60: 1-0
: 1) gradient elution, elution process TLC tracing detection, merge same composition and obtain 3 sections of Fr3-1-Fr3-3, by Fr3-3 flow point warp
Silica gel column chromatography (petroleum ether-ethyl acetate system, methylene chloride-methanol gradient elution), obtains compound 6mg IS-1 repeatedly,
100mg IS-5,24mg IS-14；
By the flow point Fr.4 of step S4 gained, through silica gel (200-300 mesh) column chromatography, petroleum ether-ethyl acetate (40: 1-0
: 1) gradient elution, elution process TLC tracing detection, merge same composition and obtain 4 sections of Fr4-1-Fr4-4, by Fr4-2 flow point warp
Silica gel column chromatography (methylene chloride-methanol gradient elution) repeatedly, obtains compound 18mg IS-2,10mg IS-4；Fr4-3 is flowed
Lease making silica gel column chromatography repeatedly (methylene chloride-methanol gradient elution), and combine MCI CHP-20P (methanol-water), obtain chemical combination
Thing 18mg IS-3,10mg IS-6,14mg IS-12；
By the flow point Fr.5 of step S4 gained, through silica gel (200-300 mesh) column chromatography, methylene chloride-methanol (60: 1-1:
1) gradient elution, elution process TLC tracing detection, merge same composition and obtain 5 sections of Fr5-1-Fr5-5, by Fr5-1 flow point through instead
Multiple silica gel column chromatography (methylene chloride-methanol gradient elution), and combine Sephadex LH-20 (methanol-water) and obtain compound
18mg IS-10,10mg IS-17；By Fr5-3 flow point through silica gel column chromatography repeatedly (methylene chloride-methanol gradient elution), and tie
Close MCI CHP-20P (methanol-water), obtain compound 45mg IS-11,10mg IS-13；By Fr5-4 flow point through silica gel repeatedly
Column chromatography (methylene chloride-methanol gradient elution), purifies and obtains compound 7mg IS-15；
By the flow point Fr.6 of step S4 gained, through silica gel (200-300 mesh) column chromatography, methylene chloride-methanol (60: 1-1:
1) gradient elution, elution process TLC tracing detection, merge same composition and obtain 3 sections of Fr6-1-Fr6-3, by Fr6-3 flow point through instead
Multiple silica gel column chromatography (methylene chloride-methanol gradient elution), and combine MCI CHP-20P (methanol-water), obtain compound 14mg
IS-16。
The analysis method of above-mentioned salsola-like inula flower and herb extract, comprises the following steps:
Step one, the preparation of standard liquid
Accurately weigh above-mentioned 7 compounds 1.0mg standard items (being accurate to 0.001g) with electronic balance, use 0.2mL methyl alcohol
Dissolve, and be settled in 1.0mL volumetric flask, freezen protective.Take 10 μ L, 40 μ L, 80 μ L, 150 μ L, the 200 above-mentioned standards of μ L respectively
Solution is in 1.0mL volumetric flask, and by methanol constant volume, being configured to concentration after mixing is 10mg/L, 40mg/L, 80mg/L, 150mg/
The series standard solution of L, 200mg/L.Preserve stand-by at 4 DEG C.
Step 2, HPLC method
Table 2 HPLC analyzes method
Chromatographic column: Phenomenex Luna C-18 (250mm × 4.6mm 5um)
Column temperature: 20 DEG C
Flowing phase: A (water)-C (acetonitrile)
Flow velocity: 1mL/min
Detection wavelength: 210nm
Step 3, linear relationship examination
With the series standard solution of each compound prepared in high performance liquid chromatograph analytical table 2, sample size 6 μ L, divide
Do not record the peak area of each titer.It when drawing calibration curve, with concentration of standard solution as abscissa, with its respective peaks area is
Ordinate, calibration curve and each parameter thereof are shown in Table 3.Result shows, face, peak when each compound concentration is in the range of 10-200mg/L
Long-pending is good linear relationship with concentration, R2Value is all higher than 0.999.
37 compound standard curves of table
The preparation of need testing solution
Accurately weighing each 10.0g of two kinds of salsola-like inula flower and herb samples, adding 40mL90% ethanol, 70 ° ultrasonic extracts twice, every time
2h, merging filtrate final vacuum concentrates and is settled to 10mL, and supernatant preserves stand-by after 0.22 μm of filtering with microporous membrane at 4 DEG C.
In sample, each compounds content measures
Accurate absorption 60 μ L need testing solutions, are measured by high performance liquid chromatograph.Contrast retention time and ultraviolet
Collection of illustrative plates, determines that each monomeric compound goes out peak position in need testing solution.And record the peak area of each compound in extract,
Calibration curve method is utilized to calculate content in total plant extracts for each compound, parallel determination three times.Result of calculation is shown in Table
4。
Each compounds content in table 4 salsola-like inula flower and herb
bSolids content/sample size in recovery rate=extract.
Repellent and Antifeeding Effects to phyllotreta striolata for the compound extract is studied
Put 2 leaves (not being injured, area is equally 20 square centimeters) in same culture dish, a piece of process,
Piece compares.Being then placed in 10 phyllotreta striolatas, within 24 hours, " Invest, Then Investigate " is processed and comparison insect population number above, obtains according to difference
Repellent effect；Investigate both injured areas, obtain diet effect.Dish leaf is directly immersed in liquid and then takes out at once,
Examination worm is put into after liquid is dry.Every kind of medicament does 3 repetitions.Result show this 12 samples do not embody obvious food refusal or
Insecticidal activity.
Table 5 salsola-like inula flower and herb extract and repellent and Antifeeding Effects selectively experiment to phyllotreta striolata for the monomeric compound are ground
Study carefully
Non-selective experiment
Individually being placed in 2 culture dishes processing and compareing blade (area is 20 square centimeters), other steps are same
As selectively testing, the worm amount put is the half of selectively experiment, 5.Non-selective test result indicate that, compound 4 is right
Phyllotreta striolata has repellent activity, and the rate of walking quickly and keeping away reaches 69.99%.And compound 1 and 5 has antifeedant activity, anti-food rate is respectively
It is 47.06% and 65.22%.
Compound 4:IS-4 (1-Cyclodecene-2,5,6,9-tetrol, 1,8-dimethyl-7-(1-
Methylethyl)-, [(2 α, 5 β, 6 α, 7 α, 8 β, 9 α)])
Compound 1:IS-10 (noval chemical compound, eupatolide-II)
Compound 5:IS-5 (deoxygenates sand ground britannin)
Table 6 salsola-like inula flower and herb extract and monomeric compound are to the repellent of phyllotreta striolata and the non-selective experiment of Antifeeding Effects
Research
The foregoing is only presently preferred embodiments of the present invention, be merely illustrative for invention, and nonrestrictive.
Those skilled in the art understands, can carry out many changes to it, change, even etc. in invention claim limited range
Effect, but fall within protection scope of the present invention.This experimental study shows, the sesquiterpenoids in salsola-like inula flower and herb is it
There is the basis of insecticidal activity, especially compound 4 (1-Cyclodecene-2,5,6,9-tetrol, 1,8-dimethyl-7-
(1-methylethyl)-, [(2 α, 5 β, 6 α, 7 α, 8 β, 9 α)]), compound 1 (eupatolide-II), compound 5 (deoxygenate sand
Ground britannin) etc. insect phyllotreta striolata had significantly walk quickly and keep away and antifeedant activity, and the killing of standardized extract
Worm activity is also based on this.In addition, the present invention is for compound activity study portion, selection is insect phyllotreta striolata, but
In aforementioned background art, citation understands, salsola-like inula flower and herb Plays extract is for example viscous for the agricultural pests of other classes
Worm, diamondback moth etc., it may have equal walks quickly and keeps away and Antifeedant Effects.For the sesquiterpene lactone compounds institute in salsola-like inula flower and herb
Other the extensive biologically actives having, such as antimicrobial antiphlogistic, antitumor isoreactivity, also will fall within the scope of protection of the present invention.
Claims (6)
1. a salsola-like inula flower and herb Plays extract, it is characterised in that this extract includes following composition: Ahmedabad Rhein .B:
2.122mg/g；Ovum South America chrysanthemum element: 0.544mg/g；Sand ground britannin: 1.101mg/g；15-deoxygenates in sand ground inular flower
Ester: 0.855mg/g；4 α, 5 beta epoxide ovum South America chrysanthemum element: 0.669mg/g；Eupatolide: 11.199mg/g；Apiolin:
0.362mg/g；Total amount is 16.852mg/g.From salsola-like inula flower and herb, the chemical formula of one noval chemical compound of isolated is
C21H32O8, crystal color is white, has 21 carbon atoms in structure, including 3 quaternary carbon atoms, 11 tertiary carbon atom CH, 5
Secondary carbon CH2With 2 primary carbon CH3；Structure has a carbonyl signals δ 178.36 (C-12), four olefinic carbon atoms signal δ
139.21 (C-4) and δ 136.18 (C-1), δ 132.01 (C-5), δ 128.04 (C-10), a company oxygen anomeric carbon signals δ
103.13 (C-1 '), and δ 103.13,76.70,73.65,68.80,61.04,57.02 is one group of glucose signals；5 secondary carbon
Atom, respectively δ 67.82 (C-13), 61.04 (C-6 '), 42.07 (C-9), 38.59 (C-3) and 25.24 (C-2)；11 uncles
Carbon atom removes 6 on sugar ring, is left as δ 132.01 (C-5), 128.04 (C-10), 57.02 (C-4 '), 48.54 (C-7)
With 38.46 (C-11)；Two quaternary carbons are δ 16.45 (C-15), 13.26 (C-14)；Anomeric carbon signals δ 103.13 (C-1 ') and hydrogen
Signal δ 4.12 (H-1 ') is joined directly together.
2. the extracting method of a salsola-like inula flower and herb Plays extract and noval chemical compound, it is characterised in that comprise the steps:
S1, take dry salsola-like inula flower and herb herb 10kg, with the 90% ethanol water room temperature cold soaking three times of 6 times of volumes after pulverizing, often
Secondary 24h, filters, concentrated filtrate, obtains the dark brown paste of 0.93kg, and recycling design；
S2, by the paste of step S1 gained with suitable water-dispersible after, use dichloromethane successively, ethyl acetate extracts three respectively
Secondary, each extraction position is concentrated into medicinal extract more respectively, respectively obtains 308.8g dichloromethane extract, and 81.62g ethyl acetate extracts
Take thing, 297.0g aqueous fraction；
S3, the filter residue part continuation water-cooled by subsidy S1 gained extract and take, three times, and each 24h is concentrated into medicinal extract, obtains husky rotation
Spend water extract again；
S4, the dichloromethane layer by step S2 gained, use 300g silica gel mixed sample, rotation upper 1kg silicagel column after being evaporated, with petroleum ether-
After ethyl acetate gradient, rinse with methyl alcohol, according to TLC testing result in elution process, merge same section eluent simultaneously
Concentrate, obtain 10g Fr.1,12g flow point Fr.3,18g flow point Fr.4,10g flow point Fr.5,8g flow point Fr.6, Fr.7；
S5, the flow point of above-mentioned gained is carried out lock out operation
S51, by the flow point Fr.2 of step S4 gained, through silica gel column chromatography, with petroleum ether-ethyl acetate gradient elution, by its stroke
It is divided into 4 sections of Fr2-1-Fr2-4, by Fr2-3 flow point, through silica gel column chromatography repeatedly, petroleum ether-ethyl acetate and petroleum ether-acetone
System gradient elution, is recrystallizing repeatedly so as to obtain compound 36mg IS-7,54mg IS-8, obtains Fr2-4 through silica gel column chromatography repeatedly
To compound 15mg IS-9；
S52, by the flow point Fr.3 of step S4 gained, through silica gel column chromatography, petroleum ether-ethyl acetate gradient elution, elution process
TLC tracing detection, merges same composition and obtains 3 sections of Fr3-1-Fr3-3, by Fr3-3 flow point through silica gel column chromatography repeatedly, changed
Compound 6mg IS-1,100mg IS-5,24mg IS-14；
S53, by the flow point Fr.4 of step S4 gained, through silica gel column chromatography, petroleum ether-ethyl acetate gradient elution, elution process
TLC tracing detection, merges same composition and obtains 4 sections of Fr4-1-Fr4-4, by Fr4-2 flow point through silica gel column chromatography repeatedly, changed
Compound 18mg IS-2,10mg IS-4；By Fr4-3 flow point through silica gel column chromatography repeatedly, and combine MCI CHP-20P, changed
Compound 18mg IS-3,10mg IS-6,14mg IS-12；
S54, by the flow point Fr.5 of step S4 gained, through silica gel column chromatography, methylene chloride-methanol gradient elution, elution process TLC
Tracing detection, merges same composition and obtains 5 sections of Fr5-1-Fr5-5, by Fr5-1 flow point through silica gel column chromatography repeatedly, and combine
Sephadex LH-20 obtains compound 18mg IS-10,10mg IS-17；By Fr5-3 flow point through silica gel column chromatography repeatedly, and
In conjunction with MCI CHP-20P, obtain compound 45mg IS-11,10mg IS-13；By Fr5-4 flow point through silica gel column chromatography repeatedly,
Purifying obtains compound 7mg IS-15；
S55, by the flow point Fr.6 of step S4 gained, through silica gel column chromatography, methylene chloride-methanol gradient elution, elution process TLC
Tracing detection, merges same composition and obtains 3 sections of Fr6-1-Fr6-3, by Fr6-3 flow point through silica gel column chromatography repeatedly, and combine MCI
CHP-20P, obtains compound 14mg IS-16.
3. the extracting method of a kind of salsola-like inula flower and herb Plays extract according to claim 2, it is characterised in that described
Silica gel in step S4 is 100-200 mesh；In step S5, silica gel is 200-300 mesh.
4. the extracting method of a kind of salsola-like inula flower and herb Plays extract according to claim 2, it is characterised in that described
In step S4, the gradient of petroleum ether-ethyl acetate is 200: 1-0: 1；In described step S51, the gradient of petroleum ether-ethyl acetate is
100∶1-5∶1；In step S52, the gradient of petroleum ether-ethyl acetate is 60: 1-0: 1；Petroleum ether-ethyl acetate in step S53
Gradient is 40: 1-0: 1.
5. the extracting method of a kind of salsola-like inula flower and herb Plays extract according to claim 2, it is characterised in that described
In step S55, the gradient of methylene chloride-methanol is 60: 1-1: 1.
6. the application of a kind of salsola-like inula flower and herb Plays extract as claimed in claim 1, it is characterised in that for yellow knee
The repellent desinsection of the insects such as flea beetle.
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|---|---|---|---|---|
| CN106619611A (en) * | 2016-10-21 | 2017-05-10 | 新疆维吾尔自治区药物研究所 | Application of eupatolide for preparing liver cancer and colon cancer resisting medicine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101747153A (en) * | 2010-01-21 | 2010-06-23 | 上海交通大学 | Inula salsoloides extract and extraction method and usage thereof |
| CN104789363A (en) * | 2015-04-23 | 2015-07-22 | 宁夏大学 | Preparation method of inula salsoloides extract and application of inula salsoloides extract as grape preservative |
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2016
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101747153A (en) * | 2010-01-21 | 2010-06-23 | 上海交通大学 | Inula salsoloides extract and extraction method and usage thereof |
| CN104789363A (en) * | 2015-04-23 | 2015-07-22 | 宁夏大学 | Preparation method of inula salsoloides extract and application of inula salsoloides extract as grape preservative |
Non-Patent Citations (11)
| Title |
|---|
| F. JESKE ET AL.: "FURTHER SESQUITERPENE LACTONES FROM INULA SALSOLOIDES", 《PHYTOCHEMISTRY》 * |
| JABER S. MOSSA ET AL.: "Sesquiterpene Lactones and Thymol Esters from Vicoa pentanema", 《J. NAT. PROD.》 * |
| NAISHENG BAI ET AL.: "Sesquiterpene Lactones from Inula britannica and Their Cytotoxic and Apoptotic Effects on Human Cancer Cell Lines", 《J. NAT. PROD.》 * |
| XIAO-JIA HUA ET AL.: "Two New Sesquiterpenes from Inula salsoloides and Their Inhibitory Activities against NO Production", 《HELVETICA CHIMICA ACTA》 * |
| YONG-MING ZHAO ET AL.: "Chemical Constituents of Plants from the Genus Inula", 《CHEMISTRY & BIODIVERSITY》 * |
| 扈晓佳: "四种药用植物的化学成分及活性研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 李玉平 等: "25种菊科植物提取物对3种植物病原菌的药效试验", 《西北农林科技大学学报(自然科学版)》 * |
| 李玉平 等: "几种菊科植物杀菌活性的初步研究", 《西北农林科技大学学报(自然科学版)》 * |
| 林坤 等: "蓼子朴乙醇提取物对小菜蛾的生物活性研究", 《中国农学通报》 * |
| 耿红梅 等: "旋覆花属植物黄酮类化合物的研究概况", 《时珍国医国药》 * |
| 陈杰 等: "沙旋复花中的化学成分", 《植物学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106619611A (en) * | 2016-10-21 | 2017-05-10 | 新疆维吾尔自治区药物研究所 | Application of eupatolide for preparing liver cancer and colon cancer resisting medicine |
| CN106619611B (en) * | 2016-10-21 | 2020-01-10 | 新疆维吾尔自治区药物研究所 | Application of eupatorium lactone in preparing medicine for resisting liver cancer and colon cancer |
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