CN103884802B - A kind of memory loss shellfish poison molecular engram integral column and application thereof - Google Patents
A kind of memory loss shellfish poison molecular engram integral column and application thereof Download PDFInfo
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- CN103884802B CN103884802B CN201210554663.9A CN201210554663A CN103884802B CN 103884802 B CN103884802 B CN 103884802B CN 201210554663 A CN201210554663 A CN 201210554663A CN 103884802 B CN103884802 B CN 103884802B
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Abstract
The present invention relates to a kind of preparation method and application of memory loss property shellfish poison molecular engram integral column.By the template molecule of suitable proportioning, function monomer, crosslinking chemical, pore-foaming agent (lauryl alcohol, 1,4-butylene glycol) be dissolved in dimethyl sulfoxide (DMSO), ultrasonic mixing 5 minutes, leave standstill 20 minutes, add initiating agent (azoisobutyronitrile) and account for 0.5 ~ 1% of monomer total amount, fill nitrogen deoxygenation 5 minutes, this solution is full of quartz capillary, end-blocking, 60 ~ 70 DEG C of constant temperature are polymerized 12 ~ 14 hours, kapillary is connected on high performance liquid chromatography infusion pump, cleaning removing template molecule and pore-foaming agent, finally obtain the capillary monolithic column with good separation effect.Molecular engram integral column prepared by the present invention has good selective recognition and little resistance to mass tranfer, as high performance liquid chromatography microtrabeculae, can realize the separation to domoic acid in shellfish samples, enrichment and detection.
Description
Technical field
The invention belongs to Polymer materialspreparation technology and the application in saxitoxin compartment analysis thereof, relate to a kind of molecular engram integral column of novel saxitoxin specifically.
Background technology
Molecular imprinting refers to preparation and application technology specific target molecules (template molecule) and analogue thereof to the polymkeric substance of specific recognition.Molecularly imprinted polymer (Molecularlyimprintedpolymers, MIPs) preparation generally adopts mass polymerization, the polymerizate obtained fully washs the materials such as removing template molecule, pore-foaming agent and unreacted monomer after grinding, sieving under certain solvent condition, and then obtains polymeric material template molecule to specific identification effect.This polymkeric substance can optionally recognition template molecule in the system of complexity, and has the advantages such as anti-adverse environment ability is strong, good stability, long service life, has been widely used in sample pre-treatments stratographic analysis.
Integral post, is also called bar-shaped post, is the continuous bed Stationary liquid that a kind of organic or inorganic polymerization carries out in-situ polymerization in chromatographic column.Integral post has the advantages such as consistent internal structure, high, the easy modification of permeability and mass transfer rate be fast, can solve the problems such as chromatography column space occupancy rate is low, mass transfer rate slow, post negative pressure is high, chromatographic peak hangover is serious, be described as the 4th generation chromatographic stationary phases.
Based on the selective recognition ability of molecularly imprinted polymer and the plurality of advantages of integral post, the two is combined and prepares the development trend that molecular engram integral column (Molecularlyimprintedmonolithiccolunln, MIPMC) becomes certainty.As a kind of novel Solid-Phase Extraction material and chromatographic stationary phases, molecular engram integral column has been widely used in the fields such as environment, biology, medicine analysis.
Summary of the invention
The object of the invention is the specific enrichment and the detection that realize memory loss class shellfish poison.A kind of preparation method of class saxitoxin integral post of losing memory is provided, and the fast enriching of class shellfish poison of losing memory in high performance liquid chromatography and detection.
In capillary column, prepare molecular engram integral column, and in liquid chromatography, achieve fast enriching and the detection of memory loss class shellfish poison, the method comprises the following steps:
(1) be dissolved in dimethyl sulfoxide solution by domoic acid standard items, add function monomer, crosslinking chemical, pore-foaming agent, ultrasonic mixing, letting nitrogen in and deoxidizing, standing at least 20min is to form stable compound.
(2) add that initiating agent accounts for total solution weight 0.5% ~ 5%, ultrasonic dissolution.
(3) (2) are poured in the quartz capillary of 25 ~ 250 μm of id modified through γ-(methacryloxy)-trimethoxy silane, end-blocking.12-16 hour is polymerized in 60 ~ 70 DEG C of water-baths
(4) trace post is connected on high performance liquid chromatography, utilizes solvent washing pore-foaming agent and template molecule, just can obtain the integral post with good imprinting effect.
(5) by high performance liquid chromatography, use trace post as trapping column and analytical column, realize fast enriching and the detection of saxitoxin.
Wherein:
Template molecule accounts for the 1-20% of function monomer gross mass;
The mass ratio of function monomer and pore-foaming agent is 1-10%
Dimethylsulfoxide solvent and pore-foaming agent volume ratio are 0.1-1.2;
Azoisobutyronitrile accounts for 0.5% ~ 5% of mixed solution gross mass.
The preparation process of the described quartz capillary column through γ-(methacryloxy)-trimethoxy silane modification is: first, adopt 0.05-0.5mol/l sodium hydrate aqueous solution to rinse kapillary 1-15 minute; Then distilled water flushing 3-15 minute is used; Then kapillary 1-15 minute is rinsed with 0.05-0.5mol/l aqueous hydrochloric acid solution, then distilled water flushing kapillary 3-15 minute is used, then kapillary 10-60 minute is blown with nitrogen, γ-(methacryloxy)-trimethoxy silane methanol solution of perfusion 0.1% is to being full of in kapillary, and in the sealing of two sections, kapillary, 60-90 DEG C of water-bath polyase 13-8 hours, then uses washed with methanol kapillary 30 minutes, then kapillary 10-60 minute is blown with 50 DEG C of nitrogen, stand-by.
Wherein be preferably:
(1) described function monomer is the potpourri of methacrylic acid, Methacrylamide, and its thing mass ratio is 10:0,9:1,8:2,7:3,6:4,5:5,4:6,3:7,2:8,1:9: or 0:10.
(2) pore-foaming agent described in is the potpourri of lauryl alcohol, BDO, and its thing mass ratio is 10:0,9:1,8:2,7:3,6:4,5:5,4:6,3:7,2:8,1:9: or 0:10.
(3) shared by described template molecule, the ratio of monomer quality is 1%, 2%, 5%, 10%, 20%, capillary column through modifying through γ-(methacryloxy)-trimethoxy silane used is 25 μm, 50 μm, 75 μm, 100 μm, 150 μm, 200 μm, 250 μm, polymerization temperature is 60 ~ 70 DEG C of water-baths, trace integral post length 5cm-20cm.
The invention has the advantages that: the molecular engram integral column having prepared memory loss class shellfish poison in quartz capillary column, analysis speed is fast, and solvent load is few, and molecular recognition performance is good.
Accompanying drawing explanation
Fig. 1 is embodiment 1 integral post scanning electron microscope (SEM) photograph.
Fig. 2 is embodiment 1 trace integral post post pressure and flow velocity relation figure (good linear relationship describes integral material consistent internal structure).
Fig. 3 is the saxitoxin (APS) extracted in the reservation murine toxin that embodiment 1 molecular blotting column (MIP) more non-molecular blotting column (NIP) can not only be stronger, the saxitoxin (APS) extracted in murine toxin can also be carried out chiral resolution, degree of separation (R)=2.1.
Embodiment
Example 1
(1) first by template molecule domoic acid 0.1 μm of ol, Methacrylamide 1.5 μm of ol, methacrylic acid 0.5 μm of ol, lauryl alcohol, 1,4-butylene glycol 17mg, piperazine bisacrylamide 1.1 μm of ol, dimethyl sulfoxide is solvent, azoisobutyronitrile 0.04mg, ultrasonic dissolution mixes, letting nitrogen in and deoxidizing, is poured in the quartz capillary of the 25 μm of internal diameters modified through γ-(methacryloxy)-trimethoxy silane, end-blocking.Be polymerized 12 hours in 60 ~ 70 DEG C of water-baths.After polyreaction completes, employing high pressure liquid chromatography pump rinses unreacted monomer and template molecule.
Example 2
(2) other are with embodiment 1, can obtain MIP2 to add pore-foaming agent (lauryl alcohol, BDO, mass ratio 10:0).
Example 3
(3) other are with embodiment 1, can obtain MIP3 to add pore-foaming agent (lauryl alcohol, BDO, mass ratio 9:1).
Example 4
(4) other are with embodiment 1, can obtain MIP4 to add pore-foaming agent (lauryl alcohol, BDO, mass ratio 8:2).
Example 5
(5) other are with embodiment 1, can obtain MIP5 to add pore-foaming agent (lauryl alcohol, BDO, mass ratio 7:3).
Example 6
(6) other are with embodiment 1, can obtain MIP6 to add pore-foaming agent (lauryl alcohol, BDO, mass ratio 6::4).
Example 7
(7) other are with embodiment 1, can obtain MIP7 to add pore-foaming agent (lauryl alcohol, BDO, mass ratio 5:5).
Claims (9)
1. a memory loss shellfish poison molecular engram integral column, is characterized in that:
Described molecular engram integral column take domoic acid as template molecule, Methacrylamide and/or methacrylic acid are function monomer, lauryl alcohol and/or BDO are pore-foaming agent, and dimethyl sulfoxide is solvent, azoisobutyronitrile is initiating agent, piperazine bisacrylamide is crosslinking chemical, and ultrasonic dissolution mixes, letting nitrogen in and deoxidizing, be poured in the quartz capillary of 25 ~ 250 μm of internal diameters modified through γ-(methacryloxy)-trimethoxy silane, end-blocking; 12-16 hour is polymerized in 60 ~ 70 DEG C of water-baths;
Polyreaction completes post-flush and removes unreacted function monomer and template molecule.
2. molecular engram integral column according to claim 1, is characterized in that:
The molar ratio of methacrylic acid and Methacrylamide is 10:0 to 0:10;
The mass ratio of lauryl alcohol and BDO is 10:0 to 0:10.
3. molecular engram integral column according to claim 1, is characterized in that:
The molar ratio of methacrylic acid and Methacrylamide is 10:0,9:1,8:2,7:3,6:4,5:5,4:6,3:7,2:8,1:9 or 0:10;
Template molecule accounts for 1%, 2%, 5%, 10% or 20% of function monomer gross mass;
The mass ratio of lauryl alcohol and BDO is 10:0,9:1,8:2,7:3,6:4,5:5,4:6,3:7,2:8,1:9 or 0:10.
4. molecular engram integral column according to claim 1, is characterized in that:
Molecular engram integral column, in the quartz capillary column modified through γ-(methacryloxy)-trimethoxy silane of internal diameter 25-250 μm, is prepared from, trace integral post length 5cm-10cm in 60 ~ 70 DEG C of water-baths.
5. molecular engram integral column according to claim 1, is characterized in that:
The preparation process of the described quartz capillary column through γ-(methacryloxy)-trimethoxy silane modification is: first, adopt 0.05-0.5mol/l sodium hydrate aqueous solution to rinse kapillary 1-15 minute; Then distilled water flushing 3-15 minute is used; Then kapillary 1-15 minute is rinsed with 0.05-0.5mol/l aqueous hydrochloric acid solution, then distilled water flushing kapillary 3-15 minute is used, then kapillary 10-60 minute is blown with nitrogen, γ-(methacryloxy)-trimethoxy silane methanol solution of perfusion 0.1% is to being full of in kapillary, and in kapillary sealing two ends, 60-90 DEG C of water-bath polyase 13-8 hours, then uses washed with methanol kapillary 30 minutes, then kapillary 10-60 minute is blown with 50 DEG C of nitrogen, stand-by.
6. molecular engram integral column according to claim 1, is characterized in that: after polyreaction completes, is connected in high performance liquid chromatography by the trace integral post of preparation, adopts high pressure liquid chromatography pump to rinse unreacted function monomer and template molecule.
7. an application for molecular engram integral column described in claim 1, the trace integral post of preparation is used for losing memory in shellfish samples the extraction of class shellfish poison or enrichment.
8. apply according to claim 7, it is characterized in that: the trace integral post of preparation is connected in high performance liquid chromatography, and in liquid chromatography, realize the separation of memory loss class shellfish poison, enrichment and detection.
9. apply according to claim 7 or 8, it is characterized in that: described memory loss class shellfish poison is domoic acid.
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| CN104198630B (en) * | 2014-08-28 | 2015-09-30 | 河南科技大学 | The preparation method of Amitraz molecular engram integral column and application |
| CN104759117B (en) * | 2015-03-25 | 2016-05-25 | 上海交通大学 | Domoic acid molecular imprinted solid phase extraction cartridge and its preparation method and application |
| CN106512963A (en) * | 2015-11-11 | 2017-03-22 | 大连出入境检验检疫局检验检疫技术中心 | Tetrodotoxin molecularly imprinted monolithic column preparation method, monolithic column and application of monolithic column |
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| WO2002008735A1 (en) * | 2000-07-21 | 2002-01-31 | Glaxo Group Limited | Raman technique for detecting molecular interactions in molecularly imprinted polymers |
| CN101690886A (en) * | 2009-06-19 | 2010-04-07 | 武汉大学 | Molecular imprinting monolithic column, preparation method and application thereof |
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| KR20120027894A (en) * | 2010-09-14 | 2012-03-22 | 한국해양연구원 | Antibody production and detecting method for detection of domoic acid from phytoplankton |
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| WO2002008735A1 (en) * | 2000-07-21 | 2002-01-31 | Glaxo Group Limited | Raman technique for detecting molecular interactions in molecularly imprinted polymers |
| CN101690886A (en) * | 2009-06-19 | 2010-04-07 | 武汉大学 | Molecular imprinting monolithic column, preparation method and application thereof |
| CN101852791A (en) * | 2010-05-21 | 2010-10-06 | 天津出入境检验检疫局动植物与食品检测中心 | Method for preparing norfloxacin molecular imprinted monolithic column in ionic liquid |
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