CN105348440A - Oblongifolin C molecularly imprinted polymer, and preparation method and application thereof - Google Patents
Oblongifolin C molecularly imprinted polymer, and preparation method and application thereof Download PDFInfo
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- C07C49/82—Ketones containing a keto group bound to a six-membered aromatic ring containing hydroxy groups
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Abstract
The invention discloses an Oblongifolin C molecularly imprinted polymer. The polymer is prepared by the following steps: (1) mixing 0.125 mmol of Oblongifolin C, 0.5-1 mmol of a functional monomer, 1.25-5 mmol of a crosslinking agent, 0.03-0.12 mmol of an initiator and 0.5-6 mL of a pore forming agent, conducting ultrasonic degassing, charging nitrogen, removing oxygen, sealing, and polymerizing in a constant temperature water bath at 40-80 DEG C for 12-24 h to obtain a polymer containing Oblongifolin C; and (2) grinding and sieving the polymer obtained in the step (1), conducting backflow by using 10% methanol solution of acetic acid to remove Oblongifolin C to obtain the Oblongifolin C molecularly imprinted polymer. The invention also discloses a preparation method and application of the Oblongifolin C molecularly imprinted polymer.
Description
Technical field
The invention belongs to natural medicine technical field, be specifically related to a kind of OblongifolinC molecularly imprinted polymer and its preparation method and application.
Background technology
Molecular imprinting (molecularimprintingtechnique, MIT) be also molecular templating techniques, belongs to supramolecular chemistry research category, relate to physical chemistry, polymer chemistry, the basic subject such as molecular chemistry and supramolecular chemistry.To refer to a certain specific target molecule (template molecule, microsphere) as template, prepare the process this one's share of expenses for a joint undertaking to specific selectivity polymkeric substance.Molecular imprinting efficient analysis be separated, there is more application in the field such as environmental monitoring and analysis, chiral separation, drug surveillance and analysis, sensor, artificial enzyme and artificial receptors.Multiple action point can be formed when template molecule contacts with polymer monomer, can be memorized (molecular recognition) by this effect of polymerization process, after template molecule is removed, polymkeric substance just defines and to match with template molecule sterie configuration, have the hole of multiple interaction sites, such hole will to template molecule and the selective identification of analogue thereof.The molecularly imprinted polymer prepared based on this technology has the features such as specific recognition, anti-adverse environment ability are strong, good stability, long service life, applied range.
Separate analytical technique is that molecularly imprinted polymer one of is applied the most widely, and what molecularly imprinted polymer was more in this field is for liquid chromatography or thin-layer chromatography stationary phase, is secondly as target compound in solid extracting agent concentration and separation complex sample.But in current research, be only as an inrichment, failing purified monomer compound is its larger limitation, in Separation of Natural Products application or there is larger limitation.
As Yunnan Fructus Resina garciniae peel extract, there are 2 main component OblongifolinC (OC) and GuttiferoneK (GK) in this extraction, its structural similitude.(the J.Agric.FoodChem.2008 such as GangXu, 56 (23): 11144-11150) from the pericarp (9.0kg) of Garciniayunnanensis, separation obtains compound OblongifolinC (9.375g fruit conversion gained output, yield: 0.10%), it adopts acetone soak extraction, extract is separated through silica gel column chromatography, chloroform, ethyl acetate, acetone is wash-out successively, chloroform wash-out position is crossed silicagel column with normal hexane-acetone system and is carried out gradient elution separation, elutriant obtains OblongifolinC through the anti-phase C18 column purification of preparation liquid phase again, for GuttiferoneK, after normal hexane-acetone system gradient elution, elutriant is crossed silicagel column with sherwood oil-acetone system again and is carried out gradient elution separation, after again adopt preparation liquid phase through anti-phase C18 post, with acetonitrile-0.1% acetic acid water for moving phase wash-out, purifying obtains (9.375g conversion gained output, yield: 0.10%), this separation method is comparatively loaded down with trivial details, efficiency is low.
Summary of the invention
Object of the present invention aims to provide a kind of PPAP class natural product OblongifolinC molecularly imprinted polymer and its preparation method and application.OblongifolinC molecularly imprinted polymer of the present invention can be used for purifies and separates from the plants such as Yunnan gamboge fruit and obtains OblongifolinC and GuttiferoneK.OblongifolinC molecularly imprinted polymer of the present invention has the imprinted sites matched with OC molecular structure, so it has the affinity of height to OC molecule.
Specifically, a first aspect of the present invention there is provided a kind of OblongifolinC molecularly imprinted polymer, and described polymkeric substance is prepared by following steps:
(1) 0.125mmolOblongifolinC, 0.5 ~ 1mmol function monomer, 1.25 ~ 5mmol linking agent, 0.03 ~ 0.12mmol initiator and 0.5 ~ 6mL pore-creating agent are mixed, ultrasonic degas, then nitrogen deoxygenation is filled, in 40 ~ 80 DEG C of water bath with thermostatic control polymerizations 12 ~ 24 hours after sealing, obtain the polymkeric substance containing OblongifolinC;
(2) step (1) resulting polymers is ground, sieves, with 10% acetic acid methanol solution return removing OblongifolinC, obtain OblongifolinC molecularly imprinted polymer;
Described function monomer is selected from acrylamide, 4-vinylpridine or methacrylic acid, described linking agent is selected from Ethylene glycol dimethacrylate or divinylbenzene, described initiator is selected from Diisopropyl azodicarboxylate or 2,2'-Azobis(2,4-dimethylvaleronitrile), and described pore-creating agent is selected from methyl-sulphoxide, acetonitrile, chloroform or toluene.
A second aspect of the present invention there is provided a kind of preparation method of OblongifolinC molecularly imprinted polymer, said method comprising the steps of:
(1) 0.125mmolOblongifolinC, 0.5 ~ 1mmol function monomer, 1.25 ~ 5mmol linking agent, 0.03 ~ 0.12mmol initiator and 0.5 ~ 6mL pore-creating agent are mixed, ultrasonic degas, then nitrogen deoxygenation is filled, in 40 ~ 80 DEG C of water bath with thermostatic control polymerizations 12 ~ 24 hours after sealing, obtain the polymkeric substance containing OblongifolinC;
(2) step (1) resulting polymers is ground, sieves, with 10% acetic acid methanol solution return removing OblongifolinC, obtain OblongifolinC molecularly imprinted polymer;
Described function monomer is selected from acrylamide, 4-vinylpridine or methacrylic acid, described linking agent is selected from Ethylene glycol dimethacrylate or divinylbenzene, described initiator is selected from Diisopropyl azodicarboxylate or 2,2'-Azobis(2,4-dimethylvaleronitrile), and described pore-creating agent is selected from methyl-sulphoxide, acetonitrile, chloroform or toluene.
In a preference, described method also comprises OblongifolinC molecularly imprinted polymer is placed in the step that vacuum drying oven is dried to constant weight.
Present invention also offers the application of OblongifolinC molecularly imprinted polymer in separating-purifying OblongifolinC and GuttiferoneK.
In a preference, described application comprises the following steps:
(1) above-mentioned OblongifolinC molecularly imprinted polymer dress post is done SPE post, with 30% methyl alcohol profit post;
(2) upper prop after Yunnan gamboge fruit 95% alcohol extracts 70% ~ 100% methyl alcohol, 70 ~ 100% ethanol or 70 ~ 100% acetonitriles being dissolved, the consumption of described solvent is 2 ~ 5 times of described crude extract consumption, and the mass ratio of described crude extract and described OblongifolinC molecularly imprinted polymer is 4:1 ~ 20:1;
(3) by 30% ~ 40% methanol-eluted fractions, until the impurity in crude extract described in high performance liquid chromatography detection display all wash-out is complete;
(4) use 50% methanol-eluted fractions, until the GuttiferoneK in crude extract described in high performance liquid chromatography detection display all wash-out is complete, merge elutriant, concentrated, dry, obtain GuttiferoneK;
(3) use 70% methanol-eluted fractions, until the OblongifolinC in crude extract described in high performance liquid chromatography detection display all wash-out is complete, merge elutriant, concentrated, dry, obtain OblongifolinC.
In another preference, the chromatographic condition that described high performance liquid chromatography detects is as follows:
Chromatographic column: Shim-packVP-ODScolumn, 250 × 4.6mm, 4.5 μm;
Moving phase: C: acetonitrile, D:0.1% formic acid water;
Gradient elution program: 0-15min, C:D=30:70 → 100:0,15-35min, C:D=100:0 → 100:0,35-50min, C:D=100:0 → 30:70,50-60min, C:D=30:70 → 30:70;
Flow velocity: 1.0mL/min;
Determined wavelength: 347nm.
As used in the present invention, described function monomer is preferably acrylamide; Described linking agent is preferably Ethylene glycol dimethacrylate; Described pore-creating agent is preferably methyl-sulphoxide; Described initiator is preferably Diisopropyl azodicarboxylate.
In the application method of OblongifolinC molecularly imprinted polymer provided by the invention, the described solvent for dissolving Yunnan gamboge fruit 95% alcohol extracts is preferably 80% methyl alcohol, the consumption of described solvent is preferably 5 times of described crude extract consumption, and the mass ratio of described crude extract and described OblongifolinC molecularly imprinted polymer is preferably 12:1.
The present invention take OblongifolinC as template, and preparing with the method for mass polymerization can the molecularly imprinted polymer of specific recognition OblongifolinC.And, be that the OC molecularly imprinted polymer of the present invention that Template preparation obtains can also identify other compounds with OC similar with OblongifolinC, so, utilize it to the special avidity of microsphere, OC molecularly imprinted polymer of the present invention can be applied to by the mode of gradient elution the OblongifolinC and the GuttiferoneK with its structural similitude that to purify from the plants such as Yunnan gamboge.Confirmation is studied: with OblongifolinC molecularly imprinted polymer purification OC and GK of the present invention, the loss of compound is less, and it is that yield is significantly higher than existing method from crude extract medicinal extract one step direct purification through the present inventor.
Compared to prior art, the present invention has following advantage:
(1) molecular imprinting is applied to the separation that molecular weight is greater than the natural product of 600 by the present invention first, compared with the application of other molecular imprintings in natural product, there is certain challenge, and first passage gradient elution mode and utilize the Selective recognition function isolating construction analogue compounds of molecularly imprinted polymer.
(2) of the present invention is that the molecularly imprinted polymer production cost of template molecule is low with OblongifolinC, and organic solution consumption is few, and comparatively traditional separation method environmental hazard reduces.
(3) of the present invention with OblongifolinC be the molecularly imprinted polymer of template molecule after being applied to OblongifolinC and GuttiferoneK that to purify from Yunnan gamboge 95% crude extract, by 10% acetic acid methanol eluant solution regeneration, recycling.
(4) by purification of crude extract after OblongifolinC molecularly imprinted polymer dress post in the present invention, step is simple, and efficiency is high.
The details of all respects of the present invention is able to detailed description by chapters and sections subsequently.By hereafter and the description of claim, feature of the present invention, object and advantage will be more obvious.
Accompanying drawing explanation
Fig. 1 is that the HPLC-UV of GuttiferoneK in the embodiment of the present invention 1 detects collection of illustrative plates (347nm);
Fig. 2 is that the HPLC-UV of OblongifolinC in the embodiment of the present invention 1 detects collection of illustrative plates (347nm).
Embodiment
Below in conjunction with accompanying drawing and embodiment, the invention will be further described.It should be noted that these embodiments only for explaining explanation the present invention, its content not being limited.
Embodiment 1
(1) get the raw materials ready
The each component of the present embodiment is by following consumption proportion batching: template molecule (OblongifolinC) 84mg, function monomer (acrylamide) 36mg, linking agent (Ethylene glycol dimethacrylate) 250mg, initiator (Diisopropyl azodicarboxylate) 10mg, pore-creating agent (methyl-sulphoxide) 3mL.
(2) preparation is with the molecularly imprinted polymer of template molecule OblongifolinC
The template molecule OblongifolinC got ready in step (1), acrylamide, Ethylene glycol dimethacrylate, Diisopropyl azodicarboxylate, methyl-sulphoxide are mixed, ultrasonic 10 minutes, then inflated with nitrogen deoxygenation 10 minutes, sealing, 60 DEG C of water-baths are polymerized 24 hours, obtain the molecularly imprinted polymer with template molecule OblongifolinC.
(3) molecularly imprinted polymer of template molecule is sloughed in preparation
By the grinding of the molecularly imprinted polymer with template molecule OblongifolinC obtained in step (2), cross 400 order metallic screens, with 10% acetic acid methanol solution return, till high performance liquid chromatography detection display is without template molecule, then in 60 DEG C of dryings, OblongifolinC molecularly imprinted polymer 250mg is obtained.
The application of OblongifolinC template molecule imprinted polymer in OblongifolinC and GuttiferoneK extracts:
Take 45g Yunnan gamboge fruit powder, 225mL alcohol solvent, reflux three times, each 3 hours, concentrate drying obtained 23.3g crude extract.
Get the molecularly imprinted polymer dress post of the OblongifolinC that 250mg obtains according to the method for the present embodiment, with 30% methyl alcohol profit post.Get 1.0g Yunnan gamboge fruit crude extract, with 2ml80% methyl alcohol ultrasonic dissolution upper prop, after upper prop solution flows to end, with 30% methyl alcohol drip washing, until high performance liquid chromatography detection display impurity all wash-out is complete.Use 50% methanol aqueous solution wash-out again, until high performance liquid phase detection display GuttiferoneK all wash-out is complete, merge elutriant, concentrate drying obtains the GuttiferoneK (1.0g medicinal extract conversion gained output, yield 1.0%) that 10mg purity is 85%.Use 70% methanol-eluted fractions again, until high performance liquid chromatography detection display OblongifolinC all wash-out is complete, merge elutriant, concentrate drying obtains the OblongifolinC (1.0g medicinal extract conversion gained output, yield 2.1%) that 21mg purity is 89%.
The chromatographic condition that described high performance liquid chromatography detects is as follows:
Chromatographic column: Shim-packVP-ODScolumn, 250 × 4.6mm, 4.5 μm;
Moving phase: C: acetonitrile, D:0.1% formic acid water;
Gradient elution program: 0-15min, C:D=30:70 → 100:0,15-35min, C:D=100:0 → 100:0,35-50min, C:D=100:0 → 30:70,50-60min, C:D=30:70 → 30:70;
Flow velocity: 1.0mL/min;
Determined wavelength: 347nm.
The regeneration of OblongifolinC molecularly imprinted polymer: by the molecularly imprinted polymer after above-mentioned test, with 10% acetic acid methanol solution and methyl alcohol wash-out successively, namely renewable for extraction next time.
The relative peak area data of the HPLC-UV color atlas (Fig. 1) of compound GuttiferoneK are as follows:
Table 1
| Peak number | Retention time | Area | Relative peak area % | Highly | Integral type |
| 1 | 13.059 | 4979 | 0.45 | 511 | BB |
| 2 | 14.280 | 6536 | 0.39 | 672 | BB |
| 3 | 32.717 | 56805 | 5.13 | 4107 | BB |
| 4 | 33.312 | 24826 | 2.24 | 2213 | BV |
| 5 | 33.708 | 974989 | 88.04 | 63544 | VB |
| 6 | 36.274 | 39257 | 3.54 | 2539 | BB |
The nuclear magnetic data of compound GuttiferoneK is as follows:
H
1-NMR (400MHzinCD
3and C OD)
13-NMR (101MHzinCD
3oD) data are shown in Table 2.
The H of table 2 compound GuttiferoneK
1-NMR, C
13-NMR data
The relative peak area data of the HPLC-UV color atlas (Fig. 2) of compound OblongifolinC are as follows:
Table 3
| Peak number | Retention time | Area | Relative peak area % | Highly | Integral type |
| 1 | 32.774 | 30318 | 0.74 | 2253 | BB |
| 2 | 33.331 | 30703 | 0.75 | 1744 | BV |
| 3 | 33.718 | 104607 | 2.56 | 7553 | VB |
| 4 | 36.281 | 11567 | 0.28 | 1046 | bb |
| 5 | 40.917 | 3903151 | 95.66 | 155116 | BB |
The nuclear magnetic data of compound OblongifolinC is as follows:
H
1-NMR (400MHzinCD
3and C OD)
13-NMR (101MHzinCD
3oD) data are shown in Table 4.
The H of table 4 compound OblongifolinC
1-NMR, C
13-NMR data
Embodiment 2
(1) get the raw materials ready
The each component of the present embodiment is by following consumption proportion batching: template molecule (OblongifolinC) 84mg, function monomer (4-vinylpridine) 65.4mg, linking agent (divinylbenzene) 325mg, initiator (2,2'-Azobis(2,4-dimethylvaleronitrile)) 12mg, pore-creating agent (acetonitrile) 0.5mL.
(2) preparation is with the molecularly imprinted polymer of template molecule OblongifolinC
The template molecule OblongifolinC got ready in step (1), 4-vinylpridine, divinylbenzene, 2,2'-Azobis(2,4-dimethylvaleronitrile), acetonitrile are mixed, ultrasonic 10 minutes, then inflated with nitrogen deoxygenation 10 minutes, sealing, 40 DEG C of water-baths are polymerized 12 hours, obtain the molecularly imprinted polymer with template molecule OblongifolinC.
(3) molecularly imprinted polymer of template molecule is sloughed in preparation
By the grinding of the molecularly imprinted polymer with template molecule OblongifolinC obtained in step (2), cross 400 order metallic screens, with 10% acetic acid methanol solution return, till high performance liquid chromatography detection display is without template molecule, then in 60 DEG C of dryings, molecularly imprinted polymer 268mg is obtained.
The application of OblongifolinC template molecule imprinted polymer in OblongifolinC and GuttiferoneK extracts:
Take 45g Yunnan gamboge fruit powder, 225mL alcohol solvent, reflux three times, each 3 hours, concentrate drying obtained 23.3g crude extract.
Get 268mg and fill post according to the molecularly imprinted polymer that the method for the present embodiment is obtained, with 30% methyl alcohol profit post.Get 1.5g Yunnan gamboge fruit crude extract, note with on 4.5ml90% methyl alcohol ultrasonic dissolution, after upper prop solution flows to end, with 40% methyl alcohol drip washing, until high performance liquid chromatography detection display impurity all wash-out is complete.Use 50% methanol aqueous solution wash-out again, until high performance liquid chromatography detection display GuttiferoneK is complete by whole wash-out, merge elutriant, concentrate drying obtains the GuttiferoneK (1.5g medicinal extract conversion gained output, yield 1.2%) that 18mg purity is 87%.Use 70% methanol-eluted fractions again, until high performance liquid chromatography detection display OblongifolinC is complete by whole wash-out, merge elutriant, concentrate drying obtains the OblongifolinC (1.5g medicinal extract conversion gained output, yield 2.1%) that 31mg purity is 91%.
The chromatographic condition of the high performance liquid chromatography detection of the present embodiment is with embodiment one.
The regeneration of Oblongifolin molecularly imprinted polymer: by the molecularly imprinted polymer after above-mentioned test, with 10% acetic acid methanol solution and methyl alcohol wash-out successively, namely renewable for extraction next time.
Embodiment 3
(1) get the raw materials ready
The each component of the present embodiment is by following consumption proportion batching: template molecule (OblongifolinC) 84mg, function monomer (methacrylic acid) 75mg, linking agent (Ethylene glycol dimethacrylate) 250mg, initiator (Diisopropyl azodicarboxylate) 20mg, pore-creating agent (chloroform) 5mL.
(2) preparation is with the molecularly imprinted polymer of template molecule OblongifolinC
The template molecule OblongifolinC got ready in step (1), methacrylic acid, Ethylene glycol dimethacrylate, Diisopropyl azodicarboxylate, chloroform are mixed, ultrasonic 10 minutes, then inflated with nitrogen deoxygenation 10 minutes, sealing, 40 DEG C of water-baths are polymerized 20 hours, obtain the molecularly imprinted polymer with template molecule OblongifolinC.
(3) molecularly imprinted polymer of template molecule is sloughed in preparation
By the grinding of the molecularly imprinted polymer with template molecule OblongifolinC obtained in step (2), cross 400 order metallic screens, with 10% acetic acid methanol solution return, till high performance liquid chromatography detects without template molecule, then in 60 DEG C of dryings, molecularly imprinted polymer 230mg is obtained.
The application of OblongifolinC template molecule imprinted polymer in OblongifolinC and GuttiferoneK extracts:
Take 45g Yunnan gamboge fruit powder, 225mL alcohol solvent, reflux three times, each 3 hours, concentrate drying obtained 23.3g crude extract.
Get 230mg and fill post according to the molecularly imprinted polymer that the method for the present embodiment is obtained, with 30% methyl alcohol profit post.Get 3.0g Yunnan gamboge fruit crude extract, with 12ml70% EtOH Sonicate dissolve upper prop, after upper prop solution flows to end, with 30% methyl alcohol drip washing, until high performance liquid chromatography detection display impurity all wash-out is complete.Use 50% methanol aqueous solution wash-out again, until high performance liquid chromatography detection display GuttiferoneK all wash-out is complete, merge elutriant, concentrate drying obtains the GuttiferoneK (1.0g medicinal extract conversion gained output, yield 1.0%) that 46mg purity is 88%.Use 70% methanol-eluted fractions again, until high performance liquid phase display OblongifolinC is complete by whole wash-out, merge this elutriant, concentrate drying obtains the OblongifolinC (3.0g medicinal extract conversion gained output, yield 2.2%) that 67mg purity is 95%.
The chromatographic condition of the high performance liquid chromatography detection of the present embodiment is with embodiment one.
The regeneration of Oblongifolin molecularly imprinted polymer: by the molecularly imprinted polymer after above-mentioned test, with 10% acetic acid methanol solution and methyl alcohol wash-out successively, namely renewable for extraction next time.
Embodiment 4
(1) get the raw materials ready
The each component of the present embodiment is by following consumption proportion batching: template molecule (OblongifolinC) 84mg, function monomer (acrylamide) 72mg, linking agent (divinylbenzene) 650mg, initiator (2,2'-Azobis(2,4-dimethylvaleronitrile)) 18mg, pore-creating agent (toluene) 6mL.
(2) preparation is with the molecularly imprinted polymer of template molecule OblongifolinC
The template molecule OblongifolinC got ready in step (1), acrylamide, divinylbenzene, 2,2'-Azobis(2,4-dimethylvaleronitrile), toluene are mixed, ultrasonic 10 minutes, then inflated with nitrogen deoxygenation 10 minutes, sealing, 80 DEG C of water-baths are polymerized 24 hours, obtain the molecularly imprinted polymer with template molecule OblongifolinC.
(3) molecularly imprinted polymer of template molecule is sloughed in preparation
By the grinding of the molecularly imprinted polymer with template molecule OblongifolinC obtained in step (2), cross 400 order metallic screens, with 10% acetic acid methanol solution return, till high performance liquid phase detects without template molecule, then in 60 DEG C of dryings, molecularly imprinted polymer 247mg is obtained.
The application of OblongifolinC template molecule imprinted polymer in OblongifolinC and GuttiferoneK extracts:
Take 45g Yunnan gamboge fruit powder, 225mL alcohol solvent, reflux three times, each 3 hours, concentrate drying obtained 23.3g crude extract.
Get 247mg and fill post according to the molecularly imprinted polymer that the method for the present embodiment is obtained, with 30% methyl alcohol profit post.
Get 5.0g Yunnan gamboge fruit crude extract, with 25ml acetonitrile ultrasonic dissolution upper prop, after upper prop solution flows to end, with 40% methyl alcohol drip washing, until high performance liquid chromatography detection display impurity all wash-out is complete.Use 50% methanol aqueous solution wash-out again, until high performance liquid chromatography detection display GuttiferoneK all wash-out is complete, close this elutriant, concentrate drying obtains the GuttiferoneK (5.0g medicinal extract conversion gained output, yield 1.0%) that 49mg purity is 87%.Use 70% methanol-eluted fractions again, until high performance liquid chromatography detection display OblongifolinC all wash-out is complete, merge elutriant, concentrate drying obtains the OblongifolinC (5.0g medicinal extract conversion gained output, yield 1.4%) that 71mg purity is 95%.
The chromatographic condition of the high performance liquid chromatography detection of the present embodiment is with embodiment one.
The regeneration of Oblongifolin molecularly imprinted polymer: by the molecularly imprinted polymer after above-mentioned test, with 10% acetic acid methanol solution and methyl alcohol wash-out successively, namely renewable for extraction next time.
Many aspects involved in the present invention have been done and have as above been set forth.It is to be understood, however, that put before not departing from spirit of the present invention, those skilled in the art can carry out equivalent change and modification to it, and described change and modification fall into the coverage of the application's claims equally.
Claims (6)
1. an OblongifolinC molecularly imprinted polymer, is characterized in that, described polymkeric substance is prepared by following steps:
(1) 0.125mmolOblongifolinC, 0.5 ~ 1mmol function monomer, 1.25 ~ 5mmol linking agent, 0.03 ~ 0.12mmol initiator and 0.5 ~ 6mL pore-creating agent are mixed, ultrasonic degas, then nitrogen deoxygenation is filled, in 40 ~ 80 DEG C of water bath with thermostatic control polymerizations 12 ~ 24 hours after sealing, obtain the polymkeric substance containing OblongifolinC;
(2) step (1) resulting polymers is ground, sieves, with 10% acetic acid methanol solution return removing OblongifolinC, obtain OblongifolinC molecularly imprinted polymer;
Described function monomer is selected from acrylamide, 4-vinylpridine or methacrylic acid, described linking agent is selected from Ethylene glycol dimethacrylate or divinylbenzene, described initiator is selected from Diisopropyl azodicarboxylate or 2,2'-Azobis(2,4-dimethylvaleronitrile), and described pore-creating agent is selected from methyl-sulphoxide, acetonitrile, chloroform or toluene.
2. a preparation method for OblongifolinC molecularly imprinted polymer as claimed in claim 1, is characterized in that, said method comprising the steps of:
(1) 0.125mmolOblongifolinC, 0.5 ~ 1mmol function monomer, 1.25 ~ 5mmol linking agent, 0.03 ~ 0.12mmol initiator and 0.5 ~ 6mL pore-creating agent are mixed, ultrasonic degas, then nitrogen deoxygenation is filled, in 40 ~ 80 DEG C of water bath with thermostatic control polymerizations 12 ~ 24 hours after sealing, obtain the polymkeric substance containing OblongifolinC;
(2) step (1) resulting polymers is ground, sieves, with 10% acetic acid methanol solution return removing OblongifolinC, obtain OblongifolinC molecularly imprinted polymer;
Described function monomer is selected from acrylamide, 4-vinylpridine or methacrylic acid, described linking agent is selected from Ethylene glycol dimethacrylate or divinylbenzene, described initiator is selected from Diisopropyl azodicarboxylate or 2,2'-Azobis(2,4-dimethylvaleronitrile), and described pore-creating agent is selected from methyl-sulphoxide, acetonitrile, chloroform or toluene.
3. method as claimed in claim 2, is characterized in that, described method also comprises OblongifolinC molecularly imprinted polymer is placed in the step that vacuum drying oven is dried to constant weight.
4. the application of OblongifolinC molecularly imprinted polymer in separating-purifying OblongifolinC and GuttiferoneK as claimed in claim 1.
5. apply as claimed in claim 4, it is characterized in that, described application comprises the following steps:
(1) OblongifolinC molecularly imprinted polymer dress post as claimed in claim 1 is done SPE post, with 30% methyl alcohol profit post;
(2) upper prop after Yunnan gamboge fruit 95% alcohol extracts 70% ~ 100% methyl alcohol, 70 ~ 100% ethanol or 70 ~ 100% acetonitriles being dissolved, the consumption of described solvent is 2 ~ 5 times of described crude extract consumption, and the mass ratio of described crude extract and described OblongifolinC molecularly imprinted polymer is 4:1 ~ 20:1;
(3) by 30% ~ 40% methanol-eluted fractions, until the impurity in crude extract described in high performance liquid chromatography detection display all wash-out is complete;
(4) use 50% methanol-eluted fractions, until the GuttiferoneK in crude extract described in high performance liquid chromatography detection display all wash-out is complete, merge elutriant, concentrated, dry, obtain GuttiferoneK;
(3) use 70% methanol-eluted fractions, until the OblongifolinC in crude extract described in high performance liquid chromatography detection display all wash-out is complete, merge elutriant, concentrated, dry, obtain OblongifolinC.
6. apply as claimed in claim 5, it is characterized in that, the chromatographic condition that described high performance liquid chromatography detects is as follows:
Chromatographic column: Shim-packVP-ODScolumn, 250 × 4.6mm, 4.5 μm;
Moving phase: C: acetonitrile, D:0.1% formic acid water;
Gradient elution program: 0-15min, C:D=30:70 → 100:0,15-35min, C:D=100:0 → 100:0,35-50min, C:D=100:0 → 30:70,50-60min, C:D=30:70 → 30:70;
Flow velocity: 1.0mL/min;
Determined wavelength: 347nm.
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