CN109180622A - The method of guainane type sesquiterpenoid is extracted from globe artichoke - Google Patents
The method of guainane type sesquiterpenoid is extracted from globe artichoke Download PDFInfo
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Abstract
The method that the present invention provides a kind of to extract guainane type sesquiterpenoid from globe artichoke.The method: crude extract is obtained after being extracted after globe artichoke is crushed with ethanol water；Crude extract loading to AB-8 type macroreticular resin chromatographic column is eluted, and is detected through chromatography, and the eluent containing target compound is collected, concentrated freeze-dried to obtain crude extract；Loading to RP-C18 chromatographic column is eluted after crude extract is dissolved with 50% methanol, is detected through chromatography, and the eluent containing target compound is collected, concentrated freeze-dried to obtain pre-separation object；Pre-separation object is dissolved with acetonitrile solution, liquid phase half is carried out and prepares, collect the flow point section containing target compound, it is concentrated freeze-dried to obtain quasi- extract；Quasi- extract is dissolved with methanol aqueous solution, liquid phase half is carried out and prepares, collect the flow point section containing target compound, it is concentrated freeze-dried to obtain target compound.Method provided by the present application, efficiently separates out target compound from globe artichoke, and DNA purity is high.
Description
Technical field
The present invention relates to extraction purification fields, extract guainane type sesquialter from globe artichoke in particular to one kind
The method of terpene compound.
Background technique
Globe artichoke is composite family cynara scolymus category herbaceos perennial also known as cynara scolymus, arithoke, chrysanthemum Ji, lotus lily, France hundred
It closes, scientific name Cynara scolymus L..Mediterranean, south of europe and north African are originated in, 19th century are by French be passed to
State Shanghai is now mainly distributed on the ground such as Shanghai, Zhejiang, Yunnan, Hunan, Shandong and Beijing.Globe artichoke edible value is high, has
The good reputation of " emperors of vegetables ".
Artichoke leaf extract is used for always folk medicine for a long time, and external and clinical test shows that globe artichoke mentions
Take object have anti-oxidant, antibacterial, anticancer is antitumor, liver protection, it is hypoglycemic, improve digestion and improve hypercholesterolemia and other effects,
Half times of terpene lactones of guainane is expected to be developed as treatment diseases associated with inflammation (such as rheumatism joint as its main composition
It is scorching), specificity anti-reflecting B type B virus, the relevant novel PTP-1B inhibitor of type-2 diabetes mellitus, angiogenesis inhibitors and control
It treats the novel drugs of kinds cancer and promotes WeiDongLi Capsule and treat the novel drugs of gastric ulcer.
But the prior art for its effective component isolate and purify research it is limited.How to extract from globe artichoke and more creates
The bioactive ingredients such as wooden half times of terpene compound of alkane type, it is of crucial importance to deep development globe artichoke crop.
In view of this, the present invention is specifically proposed.
Summary of the invention
The method that the purpose of the present invention is to provide a kind of to extract guainane type sesquiterpenoid from globe artichoke, from
Target compound, and fast and effective, purity is high are extracted in globe artichoke.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
A method of it extracting guainane type sesquiterpenoid from globe artichoke, the described method comprises the following steps:
A. crude extract is obtained after being extracted after globe artichoke being crushed with ethanol water；
B. the crude extract loading is to AB-8 type macroreticular resin chromatographic column, successively with water, volume fraction 20%, 50%,
80% ethanol water is eluted, and is detected through chromatography, and the eluent containing target compound is collected, concentrated freeze-dried to obtain slightly
Extract；
C. loading is successively with volume fraction to RP-C18 chromatographic column after the crude extract is dissolved with methanol aqueous solution
30%, 48% and 100% methanol aqueous solution carries out gradient elution, detects through chromatography, collects containing the target compound
Eluent, it is concentrated freeze-dried to obtain pre-separation object；
D. the pre-separation object is dissolved with acetonitrile solution, is using Venusil MP C18 chromatographic column, with mobile phase A
Acetonitrile, Mobile phase B are aqueous formic acid as mobile phase and carry out half preparation of first time liquid phase, collect the stream containing target compound
Segmentation, it is concentrated freeze-dried to obtain quasi- extract；
E. the quasi- extract is dissolved with methanol aqueous solution, using the Venusil MP C18 chromatographic column, to flow
Phase C is methanol, mobile phase D is aqueous formic acid as mobile phase and carries out second of liquid phase, half preparation, collects and contains target chemical combination
The flow point section of object, it is concentrated freeze-dried to obtain the target compound；The structural formula of the target compound are as follows:
It is made using extraction-AB-8 type macroporous resin column chromatography-RP-C18 column chromatography-acetonitrile/aqueous formic acid liquid phase half
Standby-methanol/semipreparative the method for aqueous formic acid liquid phase, effectively separates target compound from globe artichoke.
Preferably, the production method of the AB-8 type macroreticular resin chromatographic column are as follows: first use AB-8 type macroreticular resin particle
Soaked in absolute ethyl alcohol 20-24h is pre-processed, and column is then filled, and is rinsed with water to no alcohol taste.
Pretreatment is the separating effect in order to effectively ensure macroporous resin column chromatography, avoids the interference of impurity.
Preferably, the RP-C18 chromatographic column the preparation method comprises the following steps: by RP-C18 particle be put into methanol carry out ultrasound at
Reason, and be stirred continuously, remove bubble removing；Then the RP-C18 particle and methanol handled well are filled into column together.
It is further preferred that the specification of the RP-C18 particle are as follows: 40-60 μm,。
C18 chromatographs the selection of column packing and preparation method, also for target compound is maximum from complicated ingredient
Degree is effectively separated.The impurity that filler can be effectively removed with methanol ultrasonic treatment filler, removes bubble removing, guarantees column
The effect of chromatography.
Preferably, the specification of the Venusil MP C18 chromatographic column are as follows: 10mm × 250mm, 5 μm.
It is further preferred that the volume fraction of aqueous formic acid is 0.1% in the Mobile phase B.
It is further preferred that the semipreparative gradient of first time liquid phase are as follows: initial -40 minutes, the mobile phase A with
The volume ratio of the Mobile phase B is 18:82；- 55 minutes 45 minutes, the volume ratio of the mobile phase A and the Mobile phase B was
100:0。
Preferably, the volume fraction of aqueous formic acid is 0.1% in the mobile phase D.
It is further preferred that the semipreparative gradient of second of liquid phase are as follows: -40 minutes initial, the mobile phase C
Volume ratio with the mobile phase D is 42:58；- 55 minutes 45 minutes, the volume ratio of the mobile phase C and the mobile phase D was
100:0。
Suitable chromatographic column, mobile phase and gradient, can be fast and efficiently by the target compound in complicated ingredient
It is further separated out and, so that separation becomes efficiently controllable.
Optionally, in the step A: the solid-liquid ratio of the globe artichoke and the ethanol water is 1:8-10, the second
The volume fraction of alcohol solution is 70%, extraction time 70-72h.
The optimization of leach extraction method is on the one hand to improve and extracts yield, on the other hand to guarantee the selectivity for having certain,
The structure of target compound can't be destroyed simultaneously.
Compared with prior art, the invention has the benefit that
(1) target compound has been isolated from globe artichoke；
(2) extracting method provided by the present application is simple, effective, is suitable for promoting and applying.
(3) target compound DNA purity is high.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described.
Fig. 1 is the spectrogram that embodiment 1 carries out that first time liquid phase half prepares preparation；
Fig. 2 is the spectrogram that embodiment 1 carries out that second of liquid phase half prepares preparation；
Fig. 3 is the purity testing figure for the target compound that embodiment 1 is prepared；
Fig. 4 is the mass spectrogram for the target compound that the application is prepared；
Fig. 5 is the H spectrogram for the target compound that the application is prepared；
Fig. 6 is the C spectrogram for the target compound that the application is prepared；
Fig. 7 is the H-H COSY spectrogram for the target compound that the application is prepared；
Fig. 8 is the hsqc spectrum figure for the target compound that the application is prepared；
Fig. 9 is the HMBC spectrogram for the target compound that the application is prepared.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
Globe artichoke is shone dry doubling to be crushed with pulverizer, at room temperature, globe artichoke powder is pressed into solid-liquid ratio 1:10
It is added in the ethanol water that volume fraction is 70% and extracts 72 hours, to be stirred continuously during extraction, then extracting solution is first used
Filter cloth vacuum filtration, then be filtered by vacuum with filter paper, filtrate decompression is concentrated to give crude extract.The solid content of crude extract is 0.1g/mL.
For 24 hours with soaked in absolute ethyl alcohol by AB-8 type macroreticular resin, it must be stirred continuously during immersion.Later, by resin particle
It is filled together with dehydrated alcohol column (column volume 1L), and is washed till efflux with dehydrated alcohol and adds water (1:5) not muddy, then use pure water
It rinses to no alcohol taste.By the pure water in pillar put to it is equal with resin when, 1L crude extract is slowly added into chromatographic column, arrive column
The resin of sub- lower layer starts to receive the efflux (light green color, brown color) for having obvious color, feed liquor phase after all being caught color
Analysis, in order to avoid sample is excessive, macroporous resin adsorption is incomplete, overload phenomenon occurs.After sample is adsorbed completely, successively use
The second that the ethanol water and volume fraction that ethanol water that water, volume fraction are 20%, volume fraction are 50% are 80%
Alcohol solution is eluted, and is eluted to colourless (constant always), while with clean vial picking out flow point (every 400mL connects one
Flow point).The flow point picked out is numbered as A1, A2, A3 ....The flow point of collection is crossed into 0.45 μm of membrane filtration, is then carried out high
Effect liquid phase chromatogram detection.Liquid chromatographic detection condition: column model is Venusil MP C18,4.6mm × 250mm, 5 μm；
Detection wavelength is 205nm, and mobile phase A is methanol, and Mobile phase B is 0.1% formic acid water, 20 μ L of sample volume.Gradient are as follows: just
Begin, the volume ratio of mobile phase A and Mobile phase B is 5:95；It is 100% methanol to variation in 30 minutes.It is closed according to testing result
And active component A 27-29 is selected further to separate and purify, it is concentrated freeze-dried to obtain crude extract.
By RP-C18 filler (40-60 μm,) be put into methanol and be ultrasonically treated, it is needed not in ultrasonic procedure
Disconnected stirring, removes bubble removing.Then the RP-C18 filler and methanol handled well are filled into column (column volume 1.2L) together.Crude extract is used
The methanol aqueous solution that volume fraction is 50% dissolves, then slow loading.It is successively 30% methanol, 48% methanol with volume fraction
It is eluted with 100% methanol.Eluent is received while elution, every 400mL connects a flow point, and number consecutively are as follows: C1,
C2,C3…….It is merged according to HPLC testing result, takes C11-15, be concentrated under reduced pressure after merging, freeze-drying obtains pre-separation
Object.
The acetonitrile solution that pre-separation object volume fraction is 18% is dissolved, LC3000 type high performance liquid chromatography is used
Instrument, Venusil MP C18 chromatographic column (10mm × 250mm, 5 μm) by acetonitrile, Mobile phase B of mobile phase A are volume fraction
0.1% aqueous formic acid is as mobile phase, gradient are as follows: -40 minutes initial, the mobile phase A and the Mobile phase B
Volume ratio is 18:82；- 55 minutes 45 minutes, the volume ratio of the mobile phase A and the Mobile phase B was 100:0.Control sample introduction
80 μ L, flow velocity 3mL/min, Detection wavelength 205nm are measured, first time liquid phase half is carried out and prepares, collect the stream containing target compound
It is segmented at (see Fig. 1, No. 6 peaks), it is concentrated freeze-dried to obtain quasi- extract.
The methanol aqueous solution that quasi- extract volume fraction is 50% is dissolved, LC3000 type high performance liquid chromatography is used
Instrument, Venusil MP C18 chromatographic column (10mm × 250mm, 5 μm) by methanol, mobile phase D of mobile phase C are volume fraction
0.1% aqueous formic acid is as mobile phase, gradient are as follows: and it is -40 minutes initial, the mobile phase C and mobile phase D's
Volume ratio is 42:58；- 55 minutes 45 minutes, the volume ratio of the mobile phase C and mobile phase D was 100:0, controlled sample introduction
Amount, 50 μ L, flow velocity 3mL/min, Detection wavelength 205nm carry out second of liquid phase half and prepare, collect the stream containing target compound
It is segmented at (see Fig. 2, No. 2 peaks), it is concentrated freeze-dried to obtain target compound.
Carry out purity testing to target compound: with methyl alcohol-formic acid aqueous solution, (volume ratio of methanol and aqueous formic acid is
42:58, the volume fraction of aqueous formic acid is 0.1%) to be used as mobile phase, with LC3000 type high performance liquid chromatograph, Venusil
MP C18 (4.6mm × 250mm, 5 μm), detector: UV205nm, coutroi velocity 1mL/min.Measuring purity is 97.2% (see figure
3), extracting yield is 54.5mg/kg.
Embodiment 2
Globe artichoke is shone dry doubling to be crushed with pulverizer, at room temperature, globe artichoke powder is added by solid-liquid ratio 1:8
Enter and extracted 70 hours in the ethanol water that volume fraction is 70%, is stirred continuously during extraction, then extracting solution is first with filter
Cloth vacuum filtration, then be filtered by vacuum with filter paper, filtrate decompression is concentrated to give crude extract.
By AB-8 type macroreticular resin soaked in absolute ethyl alcohol 20h, must be stirred continuously during immersion.Later, by resin particle
It is filled together with dehydrated alcohol column (column volume 1L), and is washed till efflux with dehydrated alcohol and adds water (1:5) not muddy, then use pure water
It rinses to no alcohol taste.By the pure water in pillar put to it is equal with resin when, 1L crude extract is slowly added into chromatographic column, arrive column
The resin of sub- lower layer starts to receive the efflux (light green color, brown color) for having obvious color, feed liquor phase after all being caught color
Analysis, in order to avoid sample is excessive, macroporous resin adsorption is incomplete, overload phenomenon occurs.After sample is adsorbed completely, successively use
The second that the ethanol water and volume fraction that ethanol water that water, volume fraction are 20%, volume fraction are 50% are 80%
Alcohol solution is eluted, and is eluted to colourless (constant always), while with clean vial picking out flow point (every 400mL connects one
Flow point).The flow point picked out is numbered as A1, A2, A3 ....The flow point of collection is crossed into 0.45 μm of membrane filtration, is then carried out high
Effect liquid phase chromatogram detection.Liquid chromatographic detection condition: column model is Venusil MP C18,4.6mm × 250mm, 5 μm；
Detection wavelength is 205nm, and mobile phase A is methanol, and Mobile phase B is 0.1% formic acid water, 20 μ L of sample volume.Gradient are as follows: just
Begin, the volume ratio of mobile phase A and Mobile phase B is 5:95；It is 100% methanol to variation in 30 minutes.It is closed according to testing result
And active component A 27-29 is selected further to separate and purify, it is concentrated freeze-dried to obtain crude extract.
By RP-C18 filler (40-60 μm,) be put into methanol and be ultrasonically treated, it is needed not in ultrasonic procedure
Disconnected stirring, removes bubble removing.Then the RP-C18 filler and methanol handled well are filled into column (column volume 1.2L) together.Crude extract is used
The methanol aqueous solution that volume fraction is 50% dissolves, then slow loading.It is successively 30% methanol, 48% methanol with volume fraction
It is eluted with 100% methanol.Eluent is received while elution, every 400mL connects a flow point, and number consecutively are as follows: C1,
C2,C3…….It is merged according to HPLC testing result, takes C11-15, be concentrated under reduced pressure after merging, freeze-drying obtains pre-separation
Object.
The acetonitrile solution that pre-separation object volume fraction is 18% is dissolved, LC3000 type high performance liquid chromatography is used
Instrument, Venusil MP C18 chromatographic column (10mm × 250mm, 5 μm) by acetonitrile, Mobile phase B of mobile phase A are volume fraction
0.1% aqueous formic acid is as mobile phase, gradient are as follows: -40 minutes initial, the mobile phase A and the Mobile phase B
Volume ratio is 18:82；- 55 minutes 45 minutes, the volume ratio of the mobile phase A and the Mobile phase B was 100:0.Control sample introduction
80 μ L, flow velocity 3mL/min, Detection wavelength 205nm are measured, first time liquid phase half is carried out and prepares, collect the stream containing target compound
Segmentation, it is concentrated freeze-dried to obtain quasi- extract.
The methanol aqueous solution that quasi- extract volume fraction is 50% is dissolved, LC3000 type high performance liquid chromatography is used
Instrument, Venusil MP C18 chromatographic column (10mm × 250mm, 5 μm) by methanol, mobile phase D of mobile phase C are volume fraction
0.1% aqueous formic acid is as mobile phase, gradient are as follows: and it is -40 minutes initial, the mobile phase C and mobile phase D's
Volume ratio is 42:58；- 55 minutes 45 minutes, the volume ratio of the mobile phase C and mobile phase D was 100:0, controlled sample introduction
Amount, 50 μ L, flow velocity 3mL/min, Detection wavelength 205nm carry out second of liquid phase half and prepare, collect the stream containing target compound
Segmentation, it is concentrated freeze-dried to obtain target compound.
Embodiment 3
Globe artichoke is shone dry doubling to be crushed with pulverizer, at room temperature, globe artichoke powder is added by solid-liquid ratio 1:9
Enter and extracted 71 hours in the ethanol water that volume fraction is 70%, is stirred continuously during extraction, then extracting solution is first with filter
Cloth vacuum filtration, then be filtered by vacuum with filter paper, filtrate decompression is concentrated to give crude extract.
By AB-8 type macroreticular resin soaked in absolute ethyl alcohol 22h, must be stirred continuously during immersion.Later, by resin particle
It is filled together with dehydrated alcohol column (column volume 1L), and is washed till efflux with dehydrated alcohol and adds water (1:5) not muddy, then use pure water
It rinses to no alcohol taste.By the pure water in pillar put to it is equal with resin when, 1L crude extract is slowly added into chromatographic column, arrive column
The resin of sub- lower layer starts to receive the efflux (light green color, brown color) for having obvious color, feed liquor phase after all being caught color
Analysis, in order to avoid sample is excessive, macroporous resin adsorption is incomplete, overload phenomenon occurs.After sample is adsorbed completely, successively use
The second that the ethanol water and volume fraction that ethanol water that water, volume fraction are 20%, volume fraction are 50% are 80%
Alcohol solution is eluted, and is eluted to colourless (constant always), while with clean vial picking out flow point (every 400mL connects one
Flow point).The flow point picked out is numbered as A1, A2, A3 ....The flow point of collection is crossed into 0.45 μm of membrane filtration, is then carried out high
Effect liquid phase chromatogram detection.Liquid chromatographic detection condition: column model is Venusil MP C18,4.6mm × 250mm, 5 μm；
Detection wavelength is 205nm, and mobile phase A is methanol, and Mobile phase B is 0.1% formic acid water, 20 μ L of sample volume.Gradient are as follows: just
Begin, the volume ratio of mobile phase A and Mobile phase B is 5:95；It is 100% methanol to variation in 30 minutes.It is closed according to testing result
And active component A 27-29 is selected further to separate and purify, it is concentrated freeze-dried to obtain crude extract.
By RP-C18 filler (40-60 μm,) be put into methanol and be ultrasonically treated, it is needed not in ultrasonic procedure
Disconnected stirring, removes bubble removing.Then the RP-C18 filler and methanol handled well are filled into column (column volume 1.2L) together.Crude extract is used
The methanol aqueous solution that volume fraction is 50% dissolves, then slow loading.It is successively 30% methanol, 48% methanol with volume fraction
It is eluted with 100% methanol.Eluent is received while elution, every 400mL connects a flow point, and number consecutively are as follows: C1,
C2,C3…….It is merged according to HPLC testing result, takes C11-15, be concentrated under reduced pressure after merging, freeze-drying obtains pre-separation
Object.
The acetonitrile solution that pre-separation object volume fraction is 18% is dissolved, LC3000 type high performance liquid chromatography is used
Instrument, Venusil MP C18 chromatographic column (10mm × 250mm, 5 μm) by acetonitrile, Mobile phase B of mobile phase A are volume fraction
0.1% aqueous formic acid is as mobile phase, gradient are as follows: -40 minutes initial, the mobile phase A and the Mobile phase B
Volume ratio is 18:82；- 55 minutes 45 minutes, the volume ratio of the mobile phase A and the Mobile phase B was 100:0.Control sample introduction
80 μ L, flow velocity 3mL/min, Detection wavelength 205nm are measured, first time liquid phase half is carried out and prepares, collect the stream containing target compound
Segmentation, it is concentrated freeze-dried to obtain quasi- extract.
The methanol aqueous solution that quasi- extract volume fraction is 50% is dissolved, LC3000 type high performance liquid chromatography is used
Instrument, Venusil MP C18 chromatographic column (10mm × 250mm, 5 μm) by methanol, mobile phase D of mobile phase C are volume fraction
0.1% aqueous formic acid is as mobile phase, gradient are as follows: and it is -40 minutes initial, the mobile phase C and mobile phase D's
Volume ratio is 42:58；- 55 minutes 45 minutes, the volume ratio of the mobile phase C and mobile phase D was 100:0, controlled sample introduction
Amount, 50 μ L, flow velocity 3mL/min, Detection wavelength 205nm carry out second of liquid phase half and prepare, collect the stream containing target compound
Segmentation, it is concentrated freeze-dried to obtain target compound.
LC-MS measurement and magnetic resonance detection are carried out to the compound monomer that embodiment 1-3 is obtained.
As shown in figure 4, m/z265.1475 [M+H]+, m/z287.1295 [M+Na]+, thus it is speculated that its molecular formula is C15H20O4。
The peak of H spectrum (see Fig. 5) and C spectrum (see Fig. 6) is belonged to:
1In H-NMR, the peak at δ 4.90 is water peak；Peak at δ 3.31 is methanol peak；It is two groups of dd at δ 5.00,6.14
The proton comprising there are two is learnt by integral area in peak, it is inferred that being olefinic proton signals；δ 1.19 is one group of methyl；High-Field
Partial signal should be multiple groups-CH2Group proton signal.The signal of High-Field part should be-CH2Group proton signal, and
It and include multiple groups-CH in compound2Group.13In C-NMR, the peak near δ 48 is solvent peak (deuterated methanol), is removed molten
Agent peak shares 15 groups of carbon signals in spectrogram, and the conclusion is consistent with mass spectrum conclusion.
The peak of H spectrum and C spectrum belong to as follows:
1H-NMR:2.88 (1H, m, H-1), 1.72 (1H, f, J=10.5Hz, H-2a), 2.02 (1H, m, H-2b, H-5),
3.66 (1H, m, H-3), 1.82 (1H, m, H-4), 4.02 (1H, t, J=9.5Hz, H-6), 2.78 (1H, m, H-7), 3.73 (1H,
M, H-8), 2.72 (1H, m, H-9a), 2.16 (1H, dd, J=12.5Hz, 8.9Hz, H-9b), 6.14 (2H, dd, J=25.0Hz,
3.0Hz, H-13), 5.00 (2H, d, J=16.8Hz, H-14), 1.20 (3H, d, J=6.6Hz, H-15).
13C-NMR:43.90(C-1),39.47(C-2),78.93(C-3),47.65(C-4),52.15(C-5),83.44
(C-6),54.55(C-7),74.14(C-8),46.83(C-9),145.11(C-10),140.43(C-11),172.21(C-
12),123.18(C-13),115.45(C-14),18.68(C-15)。
In conjunction with H-H COSY (see Fig. 7), HSQC (see Fig. 8), the connection position of different fragments can determine by HMBC (see Fig. 9)
Point, comprehensive all nuclear magnetic spectrograms and mass spectral results, target compound are accredited as grosheimol (guainane type sequiterpene chemical combination
Object), i.e. 3 α, 8 beta-dihydroxies-guaiaci lignum -10 (14), 11 (13)-diene β H-6 β, 12- lactone of -1 β, 3 β, 5 β, 6 α, 7.Structure
Formula are as follows:
The method provided by the present application that guainane type sesquiterpenoid is extracted from globe artichoke, separates from globe artichoke
Target compound is extracted, method simple practical is suitable for scale application, and product purity is high, for the depth of globe artichoke
Development and utilization have positive effect.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from of the invention
Many other change and modification can be made in the case where spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. a kind of method for extracting guainane type sesquiterpenoid from globe artichoke, which is characterized in that the method includes
Following steps:
A. crude extract is obtained after being extracted after globe artichoke being crushed with ethanol water；
B. the crude extract loading successively uses water, volume fraction 20%, 50%, 80% to AB-8 type macroreticular resin chromatographic column
Ethanol water carry out gradient elution, detected through chromatography, collect the eluent containing target compound, it is concentrated freeze-dried obtain it is thick
Extract；
C. loading is to RP-C18 chromatographic column after the crude extract is dissolved with methanol aqueous solution, be successively 30% with volume fraction,
48% and 100% methanol aqueous solution carries out gradient elution, detects through chromatography, collects the elution for containing the target compound
Liquid, it is concentrated freeze-dried to obtain pre-separation object；
D. the pre-separation object is dissolved with acetonitrile solution, using Venusil MP C18 chromatographic column, using mobile phase A as second
Nitrile, Mobile phase B are aqueous formic acid as mobile phase and carry out half preparation of first time liquid phase, collect the flow point containing target compound
Section, it is concentrated freeze-dried to obtain quasi- extract；
E. the quasi- extract is dissolved with methanol aqueous solution, is using the Venusil MP C18 chromatographic column, with mobile phase C
Methanol, mobile phase D are aqueous formic acid as mobile phase and carry out second of liquid phase, half preparation, collect the stream containing target compound
Segmentation, it is concentrated freeze-dried to obtain the target compound；The structural formula of the target compound are as follows:
2. the method according to claim 1, wherein the production method of the AB-8 type macroreticular resin chromatographic column
Are as follows: AB-8 type macroreticular resin particle is pre-processed with soaked in absolute ethyl alcohol 20-24h first, column is then filled, is rinsed with water to nothing
Alcohol taste.
3. the method according to claim 1, wherein the RP-C18 chromatographic column the preparation method comprises the following steps: by RP-
C18 particle, which is put into methanol, to be ultrasonically treated, and is stirred continuously, and bubble removing is removed；Then described RP-C18 will handled well
Grain and methanol fill column together.
4. according to the method described in claim 3, it is characterized in that, the specification of the RP-C18 particle are as follows: 40-60 μm,
5. the method according to claim 1, wherein the specification of the Venusil MP C18 chromatographic column are as follows:
10mm × 250mm, 5 μm.
6. according to the method described in claim 5, it is characterized in that, the volume fraction of aqueous formic acid is in the Mobile phase B
0.1%.
7. according to the method described in claim 6, it is characterized in that, the semipreparative gradient of first time liquid phase are as follows: just
Begin -40 minutes, the volume ratio of the mobile phase A and the Mobile phase B is 18:82；- 55 minutes 45 minutes, the mobile phase A with
The volume ratio of the Mobile phase B is 100:0.
8. the method according to claim 1, wherein the volume fraction of aqueous formic acid is in the mobile phase D
0.1%.
9. according to the method described in claim 8, it is characterized in that, the semipreparative gradient of second of liquid phase are as follows: just
Begin -40 minutes, the volume ratio of the mobile phase C and the mobile phase D are 42:58；- 55 minutes 45 minutes, the mobile phase C with
The volume ratio of the mobile phase D is 100:0.
10. -9 described in any item methods according to claim 1, which is characterized in that in the step A: the globe artichoke and institute
The solid-liquid ratio for stating ethanol water is 1:8-10, and the volume fraction of the ethanol water is 70%, extraction time 70-72h.
Priority Applications (1)
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109942528A (en) * | 2019-04-09 | 2019-06-28 | 湖南文理学院 | A kind of method that ionic liquid separates cynaropicrin in leaf of Cynara scolymus L |
| CN111943802A (en) * | 2020-08-19 | 2020-11-17 | 陕西源邦生物技术有限公司 | Method for preparing guaiazulene from eucalyptus globulus oil by-product |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101222931A (en) * | 2005-07-14 | 2008-07-16 | 因德纳有限公司 | Cynara scolymus extracts, the use thereof and formulations containing them |
| CN105315147A (en) * | 2014-08-03 | 2016-02-10 | 江苏康缘药业股份有限公司 | Guaiane-type sesquiterpenoids compound, and preparation method and application thereof |
| WO2016169573A1 (en) * | 2015-04-20 | 2016-10-27 | Elsebai Mahmoud Fahmi | Sesquiterpene lactones as potent and broad spectrum antiviral compounds against all genotypes of hepatitis c virus (hcv) |
-
2018
- 2018-11-30 CN CN201811457225.4A patent/CN109180622B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101222931A (en) * | 2005-07-14 | 2008-07-16 | 因德纳有限公司 | Cynara scolymus extracts, the use thereof and formulations containing them |
| CN105315147A (en) * | 2014-08-03 | 2016-02-10 | 江苏康缘药业股份有限公司 | Guaiane-type sesquiterpenoids compound, and preparation method and application thereof |
| WO2016169573A1 (en) * | 2015-04-20 | 2016-10-27 | Elsebai Mahmoud Fahmi | Sesquiterpene lactones as potent and broad spectrum antiviral compounds against all genotypes of hepatitis c virus (hcv) |
Non-Patent Citations (2)
| Title |
|---|
| GIANCARLO CRAVOTTO ET AL.: "Chemical and biological modification of cynaropicrin and grosheimin:a structure–bitterness relationship study", 《JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE》 * |
| LUCIANA SCOTTI ET AL.: "Quantitative elucidation of the structure–bitterness relationship of cynaropicrin and grosheimin derivatives", 《FOOD CHEMISTRY 》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109942528A (en) * | 2019-04-09 | 2019-06-28 | 湖南文理学院 | A kind of method that ionic liquid separates cynaropicrin in leaf of Cynara scolymus L |
| CN111943802A (en) * | 2020-08-19 | 2020-11-17 | 陕西源邦生物技术有限公司 | Method for preparing guaiazulene from eucalyptus globulus oil by-product |
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| CN109180622B (en) | 2022-09-20 |
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