CN102174148B - Preparation of triazine phytocide molecular imprinting solid phase extracting material - Google Patents
Preparation of triazine phytocide molecular imprinting solid phase extracting material Download PDFInfo
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Abstract
The invention relates to a preparation method of a triazine phytocide molecular imprinting solid phase extracting material. The method comprises the following steps of: 1. dissolving butyltripolycyanamide, alpha-methacrylic acid, glycol di(methacrylic acid)ester and azodiisobutyronitrile into trichloromethane and uniformly mixing to obtain a suspending phase I; 2. dissolving polyvinyl alcohol 1788 into water to obtain a suspending phase II; and 3. stirring and reacting the suspending phase I with the suspending phase II and drying in vacuum to obtain the triazine phytocide molecular imprinting solid phase extracting material. In the invention, by adopting a substituted molecular template, the imprinting effect is ensured and the problem of template seepage is solved; the optimized use condition is more convenient for solid phase extraction and ensures favorable impurity removing function and favorable repeatability; the matrix effect is remarkably weakened; and the triazine phytocide molecular imprinting solid phase extracting material is better than the traditional main stream solid phase extracting product, not only can ensure the detecting accuracy but also can improve the detecting sensitivity and effectively protect a chromatographic column.
Description
Technical field
The invention belongs to triazine herbicide retention analysis determination techniques field, be specifically related to pre-treatment material preparation and application in the triazine herbicide retention analysis mensuration.
Technical background
Current, along with development and the agricultural workforce of China's agricultural modernization progressively shifts to the city, the cultivation tillage method is tending towards mass-producing and intensive, and agriculture production significantly increases the demand of agricultural chemicals.Wherein weedicide rate of increase in recent years is far above sterilant and sterilant developmental level, weedicide brings in the positive benefits such as agricultural modernization and crop yield growth being widely used in of China, cause also the weeds in field resistance to be on the rise, and lure conversely the further increasing of herbicide application amount into, the result causes its residue environment and accumulating in a large number in food chain, is threatening food safety and human life healthy.Triazine herbicide is one of main weedicide kind of using at present.Triazine herbicide is synthetic take cyanuric chloride as raw material, and three chlorine atoms in the molecular structure can be by replacements such as phenol, alcohol, mercaptan, thereby generates a series of triazine herbicide.Have been reported proof, this compounds may cause human cancer and birth defects, simultaneously can disturb hormonal normal function, and because the characteristics that the triazine herbicide amount of application is large, the lasting period is long, the potential hazard that triazine herbicide causes human health and environment more and more is that people pay close attention to, the multinational Endocrine Disrupting Chemicals compound list of it having been listed in the world, each Main Agricultural state has also all formulated strict limit standard to triazine herbicide in agricultural-food and the food residual.
The research of triazine herbicide residue detection is mainly concentrated at present the mensuration of water, soil and grain sample both at home and abroad, detection method commonly used has vapor-phase chromatography, liquid phase chromatography and chromatograph-mass spectrometer coupling method etc.Because food and agricultural products retention analysis field, of a great variety, matrix is complicated, therefore in the detection analytic process of whole sample, 70% ~ 80% in addition the more time be used for the determination process.And because sample pretreatment process is complicated, expend time in longer, therefore error and mistake most possibly appear in pre-treatment step in whole testing process.At present, in the sample pretreatment process of existing triazine herbicide method for detecting residue, use more and more the Solid-Phase Extraction method.Solid phase extraction techniques is a kind of technology of extraction and fractionation analyte from solution, as a supplement or replace liquid-liquid extraction and grow up.Compare with traditional liquid-liquid extraction technology, that solid phase extraction techniques has is simple to operate, low price, loading capacity are large, easily and the advantage such as detecting instrument coupling, becomes gradually a kind of universal Sample Pretreatment Technique.The aspects such as solid phase extraction techniques its essence is a column chromatography sepn process, the selection of its separating mechanism, sorbent material and solvent and high performance liquid chromatography have many similarities.Therefore, in the factor that affects Solid-Phase Extraction efficient, the selection of sorbent material is vital influence factor.The solid phase extraction filler that is used at present the triazine herbicide retention analysis has HLB filler, neutral alumina filler, ion-exchange packing, graphitized carbon black/aminopropyl filler and graphitized carbon black/PSA filler etc.Although for some specific sample, above-mentioned filler can be removed most of impurity, for the sample of different sorts matrix complexity, it is unable to do what one wishes that the solid phase extraction filler of these non-specific adsorption just seems.Just because of this, analytical work person has introduced the molecular imprinting with high degree of specificity absorption and replaces traditional sorbent material to carry out the residual analyzing and testing of triazine herbicide as solid phase extraction filler in recent years.
Molecular imprinting (Molecular Imprinting Technology, MIT) also be the molecular template technology, is an interdisciplinary science technology that comes from the subjects such as polymer chemistry, Materials science, biological chemistry.Since at the beginning of 21 century, molecular imprinting has obtained great development as emerging branches of learning and subjects.At present, the whole world has tens countries comprising Sweden, Japan, Germany, the U.S., China, up to a hundred academic institutions and group of enterprises and institutions at least in the research and development of being engaged in molecularly imprinted polymer (molecular imprinting polymers, MIPs).The molecular imprinting development so rapidly, and is main because it has three large characteristics: structure effect precordainment, specific recognition and broad applicability.Has affinity and selectivity is high, the anti-adverse environment ability strong, good stability, long service life, the characteristics such as applied range based on the molecularly imprinted polymer of this technology preparation.Molecular imprinting has obtained studying widely and using at numerous areas such as chromatographic separation, Biomimic sensor, mimetic enzyme catalysis.MIPs with high selectivity has become a flourish new problem in the applied research aspect the sample analysis such as medicine, biochemistry and environment.The diversity of stencil-chosen, so that MIPs can be widely used in the analysis of various materials, and it also is that common solid phase extraction filler can not be compared for the high selectivity of institute's amalyzing substances.In every application of molecular imprinting, molecular engram solid phase extraction (Molecularly Imprinted Solid Phase Extraction, MISPE) be hopeful most at first to carry out business-like.Present solid phase extraction filler based on molecular imprinting there is no at home to be commercially produced, and also only has a swedish company to produce several prods abroad, but can predict, and it has very large researching value and application prospect.
The principle of molecular imprinting can simply be described as four steps:
A. function monomer and template molecule form reversible mixture under certain condition;
B. the mixing solutions of mixture and linking agent polymerization reaction take place under the conditions such as light, heat forms the highly cross-linked polymkeric substance that contains template molecule, function monomer and linking agent;
C. template molecule is removed from polymkeric substance, stayed the hole that template molecule is had recognition site and the ad hoc structure of specific selectivity at polymeric matrix;
D. after template molecule enters MIPs again, because their structure and function group all is complementary with three-dimensional structure and the recognition site in " hole ", so recognition reaction will occur, other molecules are not then because molecular volume and functional group mate and can not effectively enter " hole ", thereby they are not had recognition reaction.Obviously, action site is more in " hole ", and agency part character and type are more extensive, and then the affinity between MIPs and the template molecule and selectivity are just higher.Template molecule and analog thereof show stronger affinity interaction at MIPs, and save power is strong; Opposite other compound affinities and save power are all relatively poor.
Although the molecular engram material that existing bibliographical information synthesizes/prepares has good Selective adsorption to triazine herbicide, showed the broad prospect of application of molecular imprinting in the triazine herbicide retention analysis, but still there is outstanding problem in present research and the application, mainly contains following three aspects::
A. selected template molecule is specific triazine herbicide, therefore inevitably has the template leakage problems.Threaten even the template molecule seepage of trace also can have highly sensitive chromatograph-mass spectrometer coupling to use, cause false positive results, this also is the lower pure color spectrum analysis of more employing sensitivity and the reason of less introducing mass spectroscopy in using at present.Owing to only adopting chromatogram to carry out qualitative and quantitative analysis, therefore being difficult to satisfy the requirement of lower bound component analysis demand and confirmatory analysis method.
B. molecular imprinting mainly concentrates on soil and ambient water two class matrixes in the triazine herbicide analytical applications, this mainly is because the application surface of at present prepared triazine herbicide molecular engram material is narrow, do not carry out effective working conditions optimization when the sample that is applied to the matrix composition complexity is used, thereby at utmost do not embody the effect of molecular engram material selective adsorption.
C. existing triazine phytocide molecular imprinting solid phase extracting material adopts the bulk polymerization preparation, thereby irregular this of its profile just caused the absorption ununiformity of sorbent material, thereby cause the not withstand voltage easily in use collapse of sorbent material to cause the adsorptivity loss of energy because its physical strength is low.
Summary of the invention
In order to solve the existing defects of traditional molecular imprinting, the invention provides a kind of preparation method of new triazine phytocide molecular imprinting solid phase extracting material, and optimized the working conditions of material of the present invention by the methodological study systematicness.
Concrete technical scheme is as follows:
The preparation method of triazine phytocide molecular imprinting solid phase extracting material comprises following operation steps:
1.1 the preparation of suspending phase I
Get melamine-butyl, α-methacrylic acid (MAA), ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) and Diisopropyl azodicarboxylate (AIBN), be dissolved in trichloromethane, mix, get suspending phase I;
Wherein melamine-butyl is imprinted templates, and α-methacrylic acid (MAA) is function monomer, and ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) is linking agent, and Diisopropyl azodicarboxylate (AIBN) is initiator, and trichloromethane is solvent;
The molar ratio of imprinted templates and function monomer is 1:3, and the molar ratio of linking agent and function monomer is 4:1～6:1, and the molar ratio of initiator and function monomer is 1:4～1:6, and the volumetric usage of solvent and linking agent molar ratio are 1:2～1:4;
1.2 the preparation of suspending phase II
It is in 80～100 ℃ the hot water that polyvinyl alcohol 1788 is dissolved in temperature, then is cooled to room temperature, gets suspending phase II;
Wherein polyvinyl alcohol 1788 is protective material, and water is disperse phase; The consumption of polyvinyl alcohol 1788 is that per 50 mL water add 1 g, and the ratio of the volumetric usage of water and suspending phase I volume is 4:1～6:1;
1.3 the preparation of mixture
With slowly impouring suspending phase of suspending phase I II, and stir, get mixing solutions; 50
oUnder the C water-bath magnetic agitation condition, with 80～120 turn/rotating speed of min mixes solution reaction 6 h, continues stirring reaction 15 h under room temperature again, adopts the method collecting precipitation of filter paper filtering at the solid particulate matter of solution bottom;
1.4 remove imprinted templates and vacuum-drying
Solid particulate matter take methyl alcohol as solvent, was removed melamine-butyl (imprinted templates) and unreacted reactant in 24 hours through the Soxhlet extraction, again through 40
oC vacuum-drying gets the triazine phytocide molecular imprinting solid phase extracting material of white powder;
The consumption that each Soxhlet is extracted methyl alcohol is 200 mL.
The using method of triazine phytocide molecular imprinting solid phase extracting material comprises following operation steps:
2.1 filling and presenting of solid-phase extraction column
Taking by weighing 50 mg triazine phytocide molecular imprinting solid phase extracting materials (MIPs), to join empty capacity be in band extraction plate (aperture 20 μ m) the polypropylene solid-phase extraction column of 10 mL, then add sieve plate (aperture 20 μ m) at filler, compress with the sieve plate insertion tool;
2.2 the activation of solid-phase extraction column
Use successively 1 mL methyl alcohol, the pre-drip washing solid-phase extraction column of 1 mL water notes preventing filling surface dry in this process;
2.3 sample preextraction
Adopting first acetonitrile to extract for all kinds of samples makes extracting liquid volume less than 2 mL again and through concentrating under reduced pressure, water recovers to be settled to 10 mL again, namely get sample preextraction liquid, for the sample that contains fat, when extracting, adopt the fat in the frozen centrifugation method removal sample preextraction liquid;
2.4 loading
In above-mentioned sample preextraction process gained solution impouring post, control solution is crossed post speed and is no more than 0.5 mL/min, after solution all flows out cylinder, with 1 mL water wash, discards whole effluent liquid and drains cylinder, keeps draining continuously state 30 min;
2.5 drip washing
With 3 * 1 mL eluent methylene chloride cylinders, drain cylinder between each drip washing, discard whole leacheates;
2.6 wash-out
With 3 * 1 mL methanol-eluted fractions, the control elution flow rate is no more than 0.2 mL/min again, is evacuated to cylinder with vacuum pump between each wash-out closely dried, collects whole elutriants;
2.7 dry up and constant volume
With elutriant on Nitrogen evaporator 40
oThe C nitrogen flushing is concentrated into dried, holds again with 1 mL moving phase, and crosses 0.22 μ m organic system syringe needle filter membrane, and clear liquid is measured for liquid chromatography-tandem mass spectrometry.
Know-why of the present invention and advantage are as follows:
1. the selection of imprinted templates molecule:
The tradition molecular imprinting selects the method for template molecule and optimization imprinted templates and function monomer reaction ratio normally to select assay as template, select again different function monomers and carry out the polymerizing condition experiment according to ratios different between them, with the type of definite final template-function monomer and the molar ratio between them.This experimental technique is except taking time and effort, a defective can't avoiding is brought to such an extent that the template leakage problems is the residue problem of assay in molecularly imprinted polymer with assay as template, and this mass spectroscopy for high sensitivity is fatal.Even template molecule has obtained in theory effectively removing in the short run polymerization experiment, can't guarantee that but template molecule in the large-batch industrial finished product can obtain the removing of " perfection leaves no trace ".
Solution to this problem of the present invention is to adopt " virtual " molecular imprinting.Why be called " virtual ", refer to that analyte molecule reality do not participate in the molecular imprinting polymerization process, replace analyte to participate in the molecular imprinting polymerization process as template molecule but choose the chemical substance close with the analyte molecular structure and property.In fact, seeking the alternate template molecule is the work of a difficulty, needs the possibility of a large amount of testing data assessment alternate template molecules, not only consuming timely more needs to expend a large amount of test spendings.In order to seek suitable alternate template molecule, the modern molecular simulation analytical technology that the present invention has introduced computer supported provides foundation for the selection of alternate template molecule.
The molecular simulation analytic process is as follows:
A. alternate template Molecular Selection scope: since in the triazine herbicide molecular structure except equal total triazine rings, molecular structure all has two alkylamino group, thereby can select from the alkyl derivative of cyanuric chloride from the angle of similar;
B. the molecular simulation of alternate template molecule: the molecular structure of various alkyl derivatives uses first ChemBiodraw software (Cambridge Soft) to draw structural formula, again in Chembio3D(Cambridge Soft) use in the software MM2 algorithm to calculate least energy conformation (the parameter setting: minimum rms=0.1) of molecule, rear use MM2 molecular dynamics algorithm is proceeded molecular conformation optimization (parameter setting: step=2 fs, frame=10 fs, terminate after 10000 step), and through calculating thus the interfacial angle of molecule, the structured datas such as bond angle;
C. the molecular simulation of triazine herbicide: adopt aforesaid method to calculate the molecule of triazine herbicide, draw their structured data;
D. the initial option of alternate template molecule: carry out the T method of inspection in the molecular structure data with the molecular structure data substitution triazine herbicide of each alkyl derivative and carry out verification, forgone for the material that significant difference occurs with triazine herbicide molecular structure data.Through screening, tentatively drawing the suitable as an alternative alkyl derivative of template molecule trimeric cyanamide has: melamine-butyl, dibutyl trimeric cyanamide, tributyl trimeric cyanamide, triethyl trimeric cyanamide, triisopropyl trimeric cyanamide;
E. the further optimized choice of alternate template molecule: continue to utilize Hyperchem software (Hypercube Inc., Gainsville, FL) to carry out the molecular simulation analysis above-mentioned alternate template molecule and function monomer α-methacrylic acid.Molecular configurations adopts first MM+ molecular mechanics algorithm to be optimized, and adopts PM3 semiempirical mechanics algorithm to carry out further molecular conformation optimization again, adopts at last ab initio 3-21G Ab initio method to calculate the minimum energy of conformation;
F. possible reaction is undertaken by the Hyperchem software simulation between template molecule and the function monomer, and the calculations incorporated energy.At first adopt Amebr MM molecular mechanics algorithm that template molecule and function monomer mixture are optimized, adopt PM3 semiempirical mechanics algorithm to carry out further Optimum configuration, last ab initio 3-21G adopts the Ab initio method to calculate the minimum energy of conformation again.Template molecule all is set as 1:1 with the reaction ratio of monomer in this computation process;
The formula of calculations incorporated energy is as follows:
Δ E=[E
Possible reaction product– E
The alternate template molecule-E
Function monomer]
More easily carry out because Δ E shows more greatly the reaction of template molecule and function monomer, the reaction product character that produces is more stable.Show by top calculating, the Δ E value of melamine-butyl is maximum, therefore, selects melamine-butyl to participate in the preparation of molecularly imprinted polymer as the alternate template of triazine herbicide.
2. the reaction ratio optimization of imprinted templates molecule and function monomer: take melamine-butyl as template, change the ratio of function monomer α-methacrylic acid, continue the process by aforementioned part f, calculate the melamine-butyl of different ratios and the Δ E of function monomer α-methacrylic acid, calculation result shows that the reaction of melamine-butyl and function monomer α-methacrylic acid carries out according to the ratio of 1:2.But in order to ensure the pre-reaction effect, use function monomer according to 1.5 times of consumptions in experiment, namely reaction ratio is adjusted into 1:3;
Technical superiority: 1. adopt alternative molecular template that the Computer-aided Molecular simulation technique filters out on textural property near replaced molecule, thereby having guaranteed to use substitutes molecular template and still can reach required specific selectivity.This process approach can be saved the great many of experiments cost accurately and reliably.2. adopt alternative molecular template not only can guarantee same trace effect, more safe and reliable when using molecularly imprinted polymer to be applied to the highly sensitive mass spectrometric detection thereby the while needn't be worried to substitute the template leakage problems of molecular template.3. water molecules produces non-covalent strong bonding action when adopting virtual molecular imprinting method to avoid polymerization affects recognition site.
3. molecularly imprinted polymer prepares global optimization and the selection of polymerization reaction system
At present, the polymerization processs that use in a large number in the preparation of triazine herbicide molecular engram material are mainly take bulk polymerization as main.
The required device of mass polymerization is simple, process condition is easy to control, universality is strong, one of main method of preparation MIPs, but the method last handling process more loaded down with trivial details, yield poorly, utilization ratio, scale operation has any problem, template molecule is removed difficulty and other performance of being difficult for regulating MIPs.Therefore, the range of application of the MIPs of mass polymerization preparation is restricted.Suspension polymerization is that the used monomer of polymerization is dissolved in organic solvent, stirs in the water that is dissolved with stablizer or other intensive polar solvent high speed and forms suspension liquid, add initiator after, initiated polymerization, thus obtain the polymkeric substance of certain regular profile.
Because triazines material polarity is strong, the solubleness in water is larger, in the preextraction process as with the method for traditional saltouing, liquid-liquid extraction and anhydrous sodium sulfate drying etc. dewater, all can cause the determinand loss, and need inevitably to consider the water content factor of food and agricultural products in the sample pretreatment process, therefore it is very necessary preparing the imprinted polymer that can carry out effective molecular recognition in water (or containing water), and this will reduce the difficulty of pre-treatment preextraction process and the loss of being avoided determinand.During the molecular recognition of its material in carrying out water (or containing water) of the imprinted polymer for preparing take water as solvent medium in theory, good Selective adsorption will be had.And for the imprinted polymer polymerization process, adopting virtual molecular imprinting method can avoid when polymerization water molecules to produce non-covalent strong bonding action affects recognition site (for determinand).In order to offset the electrostatic interaction impact that produces when suspending phase is pure water, the present invention has selected to increase the concentration of function monomer α-methacrylic acid simultaneously; And in water, add polyvinyl alcohol as the reaction protective material, to reduce α-methacrylic acid and the function monomer-solubleness of template molecule reaction product in water;
Technical superiority: 1. adopt suspension polymerization process, the particle diameter of polymkeric substance can be controlled by ratio and the stirring velocity of regulating suspending phase I and suspending phase II.2. adopt water as disperse phase, can be so that prepared molecularly imprinted polymer at water or contain when carrying out molecular recognition in the material of water, have good Selective adsorption, be suitable for very much the molecular recognition process after the triazine herbicide preextraction.3. adopt polyvinyl alcohol as protective material, to reduce α-methacrylic acid and the function monomer-solubleness of template molecule reaction product in water, improved reaction yield.
4. the outward appearance of molecularly imprinted polymer characterizes
The polymkeric substance glomeration particle that suspension polymerization that the present invention adopts is prepared, record through electron microscope look polymer microballoon size distribution at 40-60 μ m, this is most sizes that are used for the filler particles of Solid-Phase Extraction purposes, because the filler of this size can be avoided high solvent resistance, also larger adsorptive power can be arranged.Resolution is of a size of 100 μ m molecularly imprinted polymer electron micrographs and sees Fig. 1;
The prepared molecularly imprinted polymer of the present invention can have been seen inner microvoid structure with height by electron microscope, and this can significantly increase the specific surface area of molecularly imprinted polymer, improves adsorptive power, and can reduce the solvent resistance.Resolution is of a size of 500 nm molecularly imprinted polymer section electron micrographs and sees Fig. 2;
Technical superiority: 1. the prepared molecularly imprinted polymer size distribution of the present invention is at 40-60 μ m, this is most sizes that are used for the filler particles of Solid-Phase Extraction purposes, because the filler of this size can be avoided high solvent resistance, also larger adsorptive power can be arranged.2. the prepared molecularly imprinted polymer of the present invention can have been seen inner microvoid structure with height by electron microscope, and this can significantly increase the specific surface area of molecularly imprinted polymer, improves adsorptive power, and can reduce the solvent resistance.
5. the optimization of molecule seal polymkeric substance working conditions
Whole extraction process and the conventional Solid-Phase Extraction of molecularly imprinted polymer during as the Solid-Phase Extraction material is similar, mainly contains to comprise pre-treatment, application of sample, the removal of impurity and 4 steps of wash-out.The present invention has carried out careful condition optimizing to each key step of molecular engram solid phase extraction.
A. the impact of adsorption solvent medium: solvent medium can have influence on the interaction between template molecule and the monomer, and therefore prediction is in carrying out the Solid-Phase Extraction process, and the adsorption solvent medium also can produce certain impact.For this reason, acetonitrile, methylene dichloride, first alcohol and water four kinds of different adsorption solvent media have been tested to the impact of absorption property.Its result shows that its adsorptivity is best when water during as adsorption medium, and this is consistent with existing bibliographical information, and the optimum solvent of namely carrying out molecular recognition is the employed solvent medium of synthetic polymer.But during take water as medium, exist certain non-specific adsorption yet, and be in the adsorption medium at methyl alcohol and acetonitrile, its absorption property is not good, and the adsorptivity of methyl alcohol is the poorest.And in methylene dichloride, not only absorption property is stronger, its non-specific adsorption also a little less than.Thereby, can select methylene dichloride as eluent, methyl alcohol is as eluent.
B. the impact of ethane nitrile content in the adsorption liquid: have a significant impact because of the absorption property of different adsorption solvent media to molecularly imprinted polymer, and owing in the sample extraction process, using acetonitrile as extraction agent, therefore be necessary to investigate in the upper prop liquid ethane nitrile content to the impact of Polymer adsorption performance.Test-results shows when the ethane nitrile content in the upper prop liquid is lower than 30%, and polymkeric substance can be finished the absorption of analyte more than 85%.Therefore, when carrying out pretreatment process, after finishing, sample extraction should remove as much as possible the acetonitrile in the extracting solution.In the actual operation, sample extracting solution in being concentrated on the Rotary Evaporators below 2 mL, and is held to 10 mL with water again, namely with the content of acetonitrile in the control upper prop liquid.
C. the consumption of sorbent material: the molecularly imprinted polymer of most literature report all have a very large loading capacity, but these loading capacities often all are the results of Staticadsorption experiment, for using molecularly imprinted polymer as the Solid-Phase Extraction material, not have more actual reference significance, the present invention has carried out the mensuration of dynamic adsorption capacity, to determine the consumption of sorbent material in the molecular engram solid phase extraction process.In view of the high sensitivity of tandem mass spectrum, and the needs of Limited Doses during practical measurement, the sample of 4 different concns measured in the test at the adsorption effect of 5 different amounts sorbent materials.Experimental result shows, when the use of sorbent material during greater than 30 mg, most determinand can be by active adsorption, and adsorption rate does not become positive correlation with the concentration of determinand, this may be because because certain action time that adsorption process needs, thereby the imprinted polymer of small amount can't be finished absorption at short notice, thereby cause penetrating of determinand, therefore in order to reach the fully absorption of determinand, answer the selected adsorbent loadings greater than 40 mg in the experiment, consider experiment effect, the selected adsorbent loadings is 50 mg in the experiment.
D. drain the impact of step: the moisture that molecular engram material remains in the cylinder in absorption determinand process from the aqueous solution can have influence on next process consumingly, thereby when using molecular engram material to carry out the molecular engram solid phase extraction process, the method that can take to drain after sample solution flows out reduces in the cylinder residual moisture to the impact of follow-up drip washing and wash-out.Thereby the present invention has carried out the time of draining to the impact of the analyte rate of recovery.Experimental result shows, in implementing the molecular engram solid phase extraction process, the moisture of thoroughly draining in the cylinder will help to improve the rate of recovery.Therefore in experimentation, select to drain cylinder 30 min.
E. the impact of eluent consumption: carried out the optimization of elution volume, namely investigate the content with determinand in the drip washing solvent streams fluid, its result shows, elution volume reaches 3 mL and occurred the determinand significantly sacrificing when above, for fear of this loss, therefore selected the elution volume of 3 mL.
F. the impact of eluting solvent consumption: the present invention has selected methyl alcohol as eluting solvent and has investigated the impact of elution volume on the rate of recovery, the result shows when elution volume reaches 3 mL can guarantee enough yields, and therefore selecting in actual use elution volume is 3 mL.
Technical superiority: each key step when the present invention uses carrying out molecular engram solid phase extraction has been carried out careful condition optimizing, can very easily molecularly imprinted polymer be used for Solid-Phase Extraction, and can guarantee under this working conditions, not only to have good removal impurity function, also have good circulation ratio.
6. the wearing quality of molecularly imprinted polymer is investigated
Although molecular engram material has the ability of specific adsorption, but what this specific character was significantly different from the immune affinitive material that has equally specific adsorption is, in case behind the employing solvent elution, the adsorptive power of immunity affinitive material can significance be lost, and as long as molecular engram material is with behind the determinand desorption, just can carry out new absorption-elution process, so the molecular engram solid phase extraction material possesses the ability that is repeatedly used.In order to investigate the wearing quality of MISPE post, the present invention has carried out repeated determination experiment.Experimental result shows reusing and still has the satisfied rate of recovery after 3 times;
Technical superiority: as long as the molecularly imprinted polymer that the present invention is prepared just can come into operation again with behind the determinand desorption, therefore uses the prepared molecularly imprinted polymer of the present invention can save the great many of experiments spending.
7. the contrast of molecularly imprinted polymer and other Solid-Phase Extraction material properties
The effect of desirable Solid-Phase Extraction in pre-treatment simultaneously is the sensitivity that the determinand in can enriched sample improves detection method, and one side is farthest to eliminate the interference that non-determinand brings.In mass spectrometric detection, the interference main manifestations that non-determinand brings is the impact of matrix effect, for the decontamination effect improving of molecularly imprinted polymer relatively and common solid-phase extraction column for the triazines material, we represent (atrazine, Atz take ten kinds of triazine herbicides as analyte; Simazine, Smz; Terbuthylazine, Tbz; Put out only Pmn; Simetryn, Smn; Dipropetryn, Dpn; Ametryn, Amn; Atraton, Ato; Te Dingtong, Tmo; Prometon Pmo), has compared Oasis HLB solid-phase extraction column (60 mg, 3 mL), N-Aluminum solid-phase extraction column (1g, 3mL), ENVI-Carb/NH
2Composite solid-phase extraction column (0.5g/0.5g, 6mL), the powers of the matrix effect of four kinds of ENVI-Carb/PSA composite solid-phase extraction columns (0.5g/0.5g, 6mL) etc. the Solid-Phase Extraction material that is mainly used in the triazine herbicide analysis when carrying out enrichment and purify representative sample group sample.The selection foundation of representative sample group and representativeness are to formulate according to the regulation of the international food code CAC of the council, and the 1 representative sample group for checking retention analysis process sees the following form
The implication of matrix effect is the percent value of adding determinand peak area in the peak area of determinand and the standard liquid in the blank extract, and this numerical value more shows that close to 1 matrix effect is more weak.Matrix effect contrast the results are shown in Table 2-14.Take table 2 as example, can find out HLB, N-aluminum, Carb/PSA and Carb/NH
2The matrix effect of four kinds of solid-phase extraction columns is larger, namely departs from 1 far, and adopts matrix effect that the MISPE method carries out sample determination by comparison closer to 1, shows and adopts MISPE can well weaken matrix effect.Table 3-14 can find out that also the molecular engram solid phase extraction material compares other Solid-Phase Extraction material and significantly weakened matrix effect.
Therefore adopt the present invention when detecting, not only can guarantee accuracy in detection, also can improve the sensitivity of detection; Also can reach in addition and effectively protect chromatographic column to prolong the purpose in the work-ing life of chromatographic column.
Matrix effect in table 2 Chinese cabbage behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 63.0% | 48.9% | 145.8% | 60.1% | 98.6% |
| Ato | 48.0% | 42.0% | 48.9% | 48.8% | 67.1% |
| Tmo | 61.3% | 21.6% | 102.2% | 17.3% | 87.1% |
| Pmn | 43.6% | 46.7% | 38.6% | 37.2% | 70.7% |
| Tbz | 45.0% | 42.5% | 120.7% | 12.0% | 93.4% |
| Pmo | 33.4% | 49.7% | 40.5% | 23.1% | 100.1% |
| Smn | 47.3% | 45.1% | 42.0% | 59.9% | 81.2% |
| Dpn | 26.1% | 22.2% | 26.0% | 24.6% | 88.2% |
| Amn | 44.4% | 81.5% | 43.6% | 48.7% | 97.3% |
| Smz | 67.3% | 34.7% | 47.1% | 87.6% | 105.2% |
Matrix effect in table 3 pears behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 54.2% | 32.7% | 66.0% | 28.6% | 95.9% |
| Ato | 44.7% | 14.0% | 36.5% | 41.3% | 86.8% |
| Tmo | 50.9% | 3.9% | 42.6% | 8.9% | 69.5% |
| Pmn | 52.5% | 15.2% | 37.7% | 51.3% | 78.8% |
| Tbz | 46.5% | 16.6% | 48.8% | 13.4% | 89.0% |
| Pmo | 46.1% | 24.1% | 51.8% | 24.9% | 94.5% |
| Smn | 55.2% | 29.2% | 51.6% | 23.6% | 91.6% |
| Dpn | 41.7% | 14.1% | 36.8% | 6.6% | 80.7% |
| Amn | 54.9% | 11.5% | 44.8% | 13.5% | 78.3% |
| Smz | 55.4% | 24.2% | 56.5% | 42.9% | 96.2% |
Matrix effect in table 4 Radix Dauci Sativae behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 55.2% | 23.6% | 41.9% | 25.8% | 69.0% |
| Ato | 43.8% | 64.5% | 29.0% | 36.8% | 73.8% |
| Tmo | 44.0% | 31.9% | 30.6% | 10.0% | 73.0% |
| Pmn | 52.6% | 57.2% | 36.6% | 48.2% | 83.4% |
| Tbz | 47.7% | 54.4% | 42.6% | 14.9% | 67.5% |
| Pmo | 55.8% | 65.3% | 47.8% | 22.5% | 93.0% |
| Smn | 52.1% | 68.2% | 41.4% | 23.3% | 95.7% |
| Dpn | 37.5% | 27.8% | 17.5% | 8.1% | 64.7% |
| Amn | 53.6% | 48.7% | 30.0% | 15.9% | 84.8% |
| Smz | 54.4% | 83.2% | 53.1% | 38.5% | 92.2% |
Matrix effect in table 5 grape behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 48.8% | 70.8% | 47.7% | 11.6% | 98.8% |
| Ato | 43.1% | 21.0% | 16.8% | 22.4% | 87.6% |
| Tmo | 26.5% | 47.9% | 28.1% | 2.1% | 88.5% |
| Pmn | 53.1% | 23.8% | 21.3% | 42.2% | 82.4% |
| Tbz | 30.7% | 37.5% | 36.3% | 3.5% | 93.3% |
| Pmo | 51.6% | 41.0% | 42.2% | 9.1% | 94.6% |
| Smn | 47.5% | 38.3% | 34.4% | 8.9% | 94.7% |
| Dpn | 15.7% | 40.9% | 37.7% | 1.6% | 88.7% |
| Amn | 36.5% | 40.8% | 39.6% | 3.9% | 87.4% |
| Smz | 43.2% | 47.2% | 41.5% | 23.6% | 99.3% |
Matrix effect in table 6 citrus behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 44.0% | 77.2% | 98.6% | 109.0% | 104.7% |
| Ato | 32.3% | 61.5% | 65.2% | 93.6% | 80.3% |
| Tmo | 35.1% | 62.0% | 75.4% | 64.7% | 80.4% |
| Pmn | 46.7% | 51.8% | 73.6% | 86.6% | 75.6% |
| Tbz | 37.3% | 61.2% | 81.5% | 67.8% | 92.0% |
| Pmo | 47.3% | 81.7% | 94.5% | 87.7% | 81.3% |
| Smn | 44.4% | 76.7% | 88.4% | 89.4% | 91.2% |
| Dpn | 25.4% | 56.3% | 70.5% | 53.3% | 81.2% |
| Amn | 41.0% | 63.6% | 80.2% | 81.0% | 81.4% |
| Smz | 47.2% | 84.5% | 91.7% | 93.0% | 106.4% |
Matrix effect in table 7 sunflower seeds behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 66.3% | 35.7% | 104.1% | 4.7% | 82.3% |
| Ato | 75.0% | 51.6% | 68.9% | 16.1% | 81.1% |
| Tmo | 66.7% | 49.5% | 73.1% | 0.8% | 70.3% |
| Pmn | 45.6% | 49.1% | 81.3% | 16.5% | 76.2% |
| Tbz | 39.0% | 38.5% | 37.6% | 1.1% | 87.6% |
| Pmo | 35.8% | 64.8% | 33.8% | 7.7% | 80.5% |
| Smn | 25.4% | 66.9% | 95.0% | 4.8% | 85.9% |
| Dpn | 22.4% | 41.9% | 57.1% | 1.1% | 73.9% |
| Amn | 46.4% | 51.1% | 79.2% | 1.6% | 75.7% |
| Smz | 42.4% | 67.8% | 95.3% | 12.8% | 86.7% |
Matrix effect behind the different Solid-Phase Extraction column purifications of table 8 rice is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 43.9% | 30.7% | 70.5% | 5.6% | 89.6% |
| Ato | 38.9% | 38.3% | 51.6% | 10.0% | 85.7% |
| Tmo | 54.1% | 8.8% | 25.8% | 0.7% | 84.9% |
| Pmn | 51.5% | 49.0% | 49.4% | 16.6% | 84.7% |
| Tbz | 21.2% | 17.1% | 39.5% | 1.4% | 87.6% |
| Pmo | 49.7% | 36.9% | 52.1% | 4.8% | 95.3% |
| Smn | 41.0% | 38.7% | 50.3% | 4.4% | 92.9% |
| Dpn | 38.0% | 5.4% | 20.3% | 0.3% | 79.7% |
| Amn | 27.0% | 24.1% | 37.9% | 2.0% | 88.6% |
| Smz | 53.8% | 53.3% | 71.9% | 9.7% | 95.3% |
Matrix effect in table 9 tealeaves behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 42.8% | 63.8% | 90.8% | 14.9% | 81.8% |
| Ato | 31.7% | 55.3% | 62.8% | 18.4% | 77.7% |
| Tmo | 21.8% | 19.8% | 41.7% | 2.5% | 73.1% |
| Pmn | 37.1% | 76.7% | 93.6% | 33.1% | 65.3% |
| Tbz | 28.7% | 26.4% | 50.1% | 5.0% | 77.2% |
| Pmo | 44.4% | 46.2% | 73.2% | 9.8% | 93.6% |
| Smn | 39.3% | 59.8% | 79.7% | 11.3% | 86.2% |
| Dpn | 19.2% | 9.9% | 21.6% | 1.6% | 78.4% |
| Amn | 34.6% | 35.3% | 62.4% | 4.1% | 72.1% |
| Smz | 46.2% | 74.9% | 90.6% | 28.3% | 81.8% |
Matrix effect in table 10 pork behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 38.3% | 81.4% | 90.9% | 7.5% | 95.4% |
| Ato | 31.9% | 51.9% | 61.3% | 12.3% | 87.0% |
| Tmo | 20.3% | 41.8% | 37.2% | 1.6% | 87.1% |
| Pmn | 44.8% | 58.2% | 54.0% | 23.5% | 86.2% |
| Tbz | 25.3% | 54.3% | 58.6% | 2.5% | 92.7% |
| Pmo | 38.3% | 67.5% | 72.8% | 6.1% | 93.5% |
| Smn | 39.2% | 67.9% | 74.7% | 5.7% | 94.9% |
| Dpn | 10.5% | 33.7% | 27.0% | 1.2% | 83.1% |
| Amn | 31.6% | 51.4% | 58.5% | 3.0% | 90.8% |
| Smz | 41.3% | 76.4% | 83.7% | 12.3% | 95.2% |
Matrix effect in table 11 pork liver behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 48.3% | 17.4% | 115.0% | 9.5% | 90.8% |
| Ato | 31.9% | 20.2% | 78.6% | 13.9% | 82.6% |
| Tmo | 22.5% | 6.1% | 85.4% | 4.0% | 60.8% |
| Pmn | 45.8% | 36.1% | 74.4% | 22.3% | 87.4% |
| Tbz | 32.6% | 5.4% | 87.3% | 5.2% | 78.4% |
| Pmo | 38.8% | 15.4% | 97.5% | 8.8% | 87.4% |
| Smn | 45.0% | 19.4% | 97.1% | 8.7% | 88.2% |
| Dpn | 19.2% | 4.1% | 75.2% | 2.9% | 60.5% |
| Amn | 36.4% | 10.8% | 87.9% | 5.4% | 73.8% |
| Smz | 44.9% | 28.7% | 98.8% | 16.0% | 91.9% |
Matrix effect in table 12 milk behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 48.7% | 31.4% | 97.3% | 8.4% | 91.2% |
| Ato | 31.9% | 28.8% | 66.4% | 13.6% | 89.7% |
| Tmo | 23.0% | 6.9% | 65.0% | 3.6% | 85.0% |
| Pmn | 46.0% | 44.7% | 71.3% | 19.2% | 92.7% |
| Tbz | 32.6% | 13.4% | 81.4% | 4.3% | 91.5% |
| Pmo | 38.8% | 24.8% | 82.9% | 8.2% | 95.3% |
| Smn | 45.0% | 31.3% | 82.9% | 7.2% | 92.1% |
| Dpn | 19.7% | 6.0% | 70.2% | 2.9% | 80.7% |
| Amn | 35.4% | 18.0% | 72.9% | 4.8% | 55.9% |
| Smz | 44.6% | 48.5% | 89.4% | 13.8% | 93.8% |
Matrix effect in table 13 egg behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 12.7% | 54.7% | 109.3% | 40.4% | 95.8% |
| Ato | 18.4% | 48.0% | 67.9% | 55.2% | 86.0% |
| Tmo | 32.3% | 19.1% | 74.7% | 12.5% | 78.5% |
| Pmn | 41.9% | 65.3% | 72.8% | 66.4% | 89.0% |
| Tbz | 35.5% | 26.8% | 81.3% | 19.9% | 80.4% |
| Pmo | 29.8% | 42.5% | 91.5% | 39.2% | 95.9% |
| Smn | 24.5% | 53.4% | 91.7% | 36.3% | 95.2% |
| Dpn | 20.8% | 11.4% | 68.2% | 8.5% | 63.1% |
| Amn | 46.4% | 29.9% | 81.6% | 23.9% | 88.9% |
| Smz | 40.6% | 68.8% | 94.3% | 55.6% | 94.1% |
Matrix effect in table 14 peeled shrimp behind the different Solid-Phase Extraction column purifications is (n=3, sample concentration: 5 μ g/kg) relatively
| The material title | HLB | N-Aluminum | Carb/PSA | Carb/NH 2 | MISPE |
| Atz | 35.5% | 59.5% | 107.8% | 6.8% | 92.1% |
| Ato | 31.3% | 48.4% | 67.5% | 13.1% | 84.0% |
| Tmo | 50.6% | 13.0% | 68.4% | 1.5% | 90.4% |
| Pmn | 42.8% | 48.4% | 74.9% | 19.6% | 90.8% |
| Tbz | 14.4% | 21.0% | 79.6% | 2.1% | 91.3% |
| Pmo | 39.9% | 37.7% | 87.3% | 5.9% | 93.0% |
| Smn | 26.5% | 47.7% | 85.8% | 4.8% | 94.2% |
| Dpn | 35.0% | 6.3% | 54.6% | 0.7% | 81.4% |
| Amn | 64.8% | 24.7% | 78.5% | 2.3% | 84.1% |
| Smz | 41.0% | 65.8% | 92.9% | 12.2% | 90.8% |
Technical superiority: 1. significantly weakened matrix effect; thereby when detecting, not only can guarantee accuracy in detection, the sensitivity 2. that also can improve detection is applicable to all vegetalitas and animality sample 3. and also can reaches in addition the purposes 4. of effectively protecting chromatographic column to prolong the work-ing life of chromatographic column and be better than present main flow Solid-Phase Extraction product aspect the matrix effect reducing.
Description of drawings
Fig. 1 is of a size of 100 μ m molecularly imprinted polymer electron micrographs for differentiating.
Fig. 2 is of a size of 500 nm molecularly imprinted polymer section electron micrographs for differentiating.
Embodiment
Below in conjunction with embodiment the present invention is further described.
Atrazine, simazine, terbuthylazine, put out clean, simetryn, dipropetryn, ametryn, atraton, Te Dingtong, prometon, Provado, pyridine worm miaow, reference material, purity is equal〉98%, available from Dr. Ehrenstorfer company (Germany); Melamine-butyl, AR, purity〉97%, available from J﹠amp; KCHEMICA(lark prestige China).Ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA), α-methacrylic acid (Methacrylic acid, MAA) is available from Sigma-Aldrich.Diisopropyl azodicarboxylate (AIBN), analytical pure is available from Tianjin Development Zone sea light chemical pharmaceutical factory.Acetonitrile, toluene, methyl alcohol, methylene dichloride, formic acid is high-efficiency liquid chromatographic-grade, available from TEDIA company (U.S.).Hydrochloric acid, primary ammonium phosphate, sodium-chlor, ammonium acetate, anhydrous sodium sulphate, trichloromethane, polyvinyl alcohol, AR, Virahol, top grade is pure, available from Chemical Reagent Co., Ltd., Sinopharm Group.
Oasis HLB solid-phase extraction column (60 mg, 3 mL) available from Waters company (U.S.), LC-Alumina-N neutral alumina solid-phase extraction column (1g, 3mL), ENVI-Carb/NH2 composite solid-phase extraction column (0.5g/0.5g, 6mL), the floride-free polypropylene SPE void column (12 mL) of polyethylene sieve plate (20 μ m) is put in advance available from the Sigma-Aldrich(U.S. in ENVI-Carb/PSA composite solid-phase extraction column (0.5g/0.5g, 6mL) and bottom).
Chinese cabbage, pears, Radix Dauci Sativae, grape, citrus, sunflower seeds, rice, tealeaves, pork, pork liver, milk, egg, peeled shrimp are all available from market.Through liquid chromatography-tandem mass spectrometry analysis confirmation negative sample.
Embodiment 1:
The preparation of triazine phytocide molecular imprinting solid phase extracting material comprises following operation steps:
1, the preparation of suspending phase I
Get melamine-butyl 0.8 g, α-methacrylic acid (MAA) 1.1 g, ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) 9.6 mL and Diisopropyl azodicarboxylate (AIBN) 0.33 g, be dissolved in 24 mL trichloromethanes, mix, get suspending phase I;
Wherein melamine-butyl is imprinted templates, and α-methacrylic acid (MAA) is function monomer, and ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) is linking agent, and Diisopropyl azodicarboxylate (AIBN) is initiator, and trichloromethane is solvent;
The molar ratio of imprinted templates and function monomer is 1:3, and the molar ratio of linking agent and function monomer is 4:1, and the molar ratio of initiator and function monomer is 1:6, and the volumetric usage of solvent and linking agent mole number ratio are 1:2.
The amount of each composition such as following table among the suspending phase I:
| The material title | Consumption |
| Melamine-butyl | 0.8 g closes 4 mmol |
| α-methacrylic acid | 1.1 g closes 12 mmol |
| Ethylene glycol bisthioglycolate (methacrylic acid) ester | 9.6 mL closes 48 mmol |
| Diisopropyl azodicarboxylate | 0.33 g closes 2 mmol |
| Trichloromethane | 24 mL |
2, the preparation of suspending phase II
It is in 80 ℃ the hot water that 2.4 g polyvinyl alcohol 1788 are dissolved in 120 mL temperature, then is cooled to room temperature, gets suspending phase II;
Wherein polyvinyl alcohol 1788 is protective material, and water is disperse phase; The consumption of polyvinyl alcohol 1788 and the volume ratio of water are 1:50, and the ratio of the volumetric usage of water and suspending phase I volume is 4:1;
3, the preparation of mixture
With slowly impouring suspending phase of suspending phase I II, and stir, get mixing solutions; 50
oUnder the C water-bath magnetic agitation condition, with 80 turn/rotating speed of min mixes solution reaction 6 h, continues stirring reaction 15 h under room temperature again, adopts the method collecting precipitation of filter paper filtering at the solid particulate matter of solution bottom;
4, remove template molecule and vacuum-drying
Solid particulate matter take methyl alcohol as solvent, was removed melamine-butyl (imprinted templates) and unreacted reactant in 24 hours through the Soxhlet extraction, again through 40
oC vacuum-drying gets the triazine phytocide molecular imprinting solid phase extracting material of white powder.
The consumption that each Soxhlet is extracted methyl alcohol is 200 mL.
Embodiment 2:
The preparation of triazine phytocide molecular imprinting solid phase extracting material comprises following operation steps:
1, the preparation of suspending phase I
Get melamine-butyl 1.5 g, α-methacrylic acid (MAA) 2.1 g, ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) 24 mL and Diisopropyl azodicarboxylate (AIBN) 0.79 g, be dissolved in the 40mL trichloromethane, mix, get suspending phase I;
Wherein melamine-butyl is imprinted templates, and α-methacrylic acid (MAA) is function monomer, and ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) is linking agent, and Diisopropyl azodicarboxylate (AIBN) is initiator, and trichloromethane is solvent;
The molar ratio of imprinted templates and function monomer is 1:3, and the molar ratio of linking agent and function monomer is 5:1, and the molar ratio of initiator and function monomer is 1:5, and the volumetric usage of solvent and linking agent mole number ratio are 1:3.
The amount of each composition such as following table among the suspending phase I:
| The material title | Consumption |
| Melamine-butyl | 1.5 g closes 8 mmol |
| α-methacrylic acid | 2.1 g closes 24 mmol |
| Ethylene glycol bisthioglycolate (methacrylic acid) ester | 24 mL close 120 mmol |
| Diisopropyl azodicarboxylate | 0.79 g closes 4.8 mmol |
| Trichloromethane | 40 mL |
2, the preparation of suspending phase II
It is in 80 ℃ the hot water that 6.2 g polyvinyl alcohol 1788 are dissolved in 310 mL temperature, then is cooled to room temperature, gets suspending phase II;
Wherein polyvinyl alcohol 1788 is protective material, and water is disperse phase; The consumption of polyvinyl alcohol 1788 and the volume ratio of water are 1:50, and the ratio of the volumetric usage of water and suspending phase I volume is 5:1;
3, the preparation of mixture
With slowly impouring suspending phase of suspending phase I II, and stir, get mixing solutions; 50
oUnder the C water-bath magnetic agitation condition, with 90 turn/rotating speed of min mixes solution reaction 6 h, continues stirring reaction 15 h under room temperature again, adopts the method collecting precipitation of filter paper filtering at the solid particulate matter of solution bottom;
4, remove template molecule and vacuum-drying
Solid particulate matter take methyl alcohol as solvent, was removed melamine-butyl (imprinted templates) and unreacted reactant in 24 hours through the Soxhlet extraction, again through 40
oC vacuum-drying gets the triazine phytocide molecular imprinting solid phase extracting material of white powder.
The consumption that each Soxhlet is extracted methyl alcohol is 200 mL.
Embodiment 3:
The preparation of triazine phytocide molecular imprinting solid phase extracting material comprises following operation steps:
1, the preparation of suspending phase I
Get melamine-butyl 1.5 g, α-methacrylic acid (MAA) 2.1 g, ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) 29 mL and Diisopropyl azodicarboxylate (AIBN) 0.99 g, be dissolved in 36 mL trichloromethanes, mix, get suspending phase I;
Wherein melamine-butyl is imprinted templates, and α-methacrylic acid (MAA) is function monomer, and ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) is linking agent, and Diisopropyl azodicarboxylate (AIBN) is initiator, and trichloromethane is solvent;
The molar ratio of imprinted templates and function monomer is 1:3, and the molar ratio of linking agent and function monomer is 6:1, and the molar ratio of initiator and function monomer is 1:4, and the volumetric usage of solvent and linking agent mole number ratio are 1:4.
The amount of each composition such as following table among the suspending phase I:
| The material title | Consumption |
| Melamine-butyl | 1.5 g closes 8 mmol |
| α-methacrylic acid | 2.1 g closes 24 mmol |
| Ethylene glycol bisthioglycolate (methacrylic acid) ester | 29 mL close 144 mmol |
| Diisopropyl azodicarboxylate | 0.99g close 6 mmol |
| Trichloromethane | 36 mL |
2, the preparation of suspending phase II
It is in 80 ℃ the hot water that 7.4 g polyvinyl alcohol 1788 are dissolved in 370 mL temperature, then is cooled to room temperature, gets suspending phase II;
Wherein polyvinyl alcohol 1788 is protective material, and water is disperse phase; The consumption of polyvinyl alcohol 1788 and the volume ratio of water are 1:50, and the ratio of the volumetric usage of water and suspending phase I volume is 6:1;
3, the preparation of mixture
With slowly impouring suspending phase of suspending phase I II, and stir, get mixing solutions; 50
oUnder the C water-bath magnetic agitation condition, with 100 turn/rotating speed of min mixes solution reaction 6 h, continues stirring reaction 15 h under room temperature again, adopts the method collecting precipitation of filter paper filtering at the solid particulate matter of solution bottom;
4, remove template molecule and vacuum-drying
Solid particulate matter take methyl alcohol as solvent, was removed melamine-butyl (imprinted templates) and unreacted reactant in 24 hours through the Soxhlet extraction, again through 40
oC vacuum-drying gets the triazine phytocide molecular imprinting solid phase extracting material of white powder.
The consumption that each Soxhlet is extracted methyl alcohol is 200 mL.
The triazine phytocide molecular imprinting solid phase extracting material application note of the present invention's preparation is as follows:
To analyze the sample in the representative sample group, namely Chinese cabbage, pears, Radix Dauci Sativae, grape, citrus, sunflower seeds, rice, tealeaves, pork, pork liver, milk, egg, peeled shrimp are example.Analyze ten kinds of modal triazine herbicides, namely atrazine, simazine, terbuthylazine, put out clean, simetryn, dipropetryn, ametryn, atraton, Te Dingtong, prometon.
1. sample preextraction
Chinese cabbage, pears, Radix Dauci Sativae, grape, citrus: in 100 mL polypropylene centrifuge tubes, take by weighing the sample after 5.0 g pulverize, add 20 mL acetonitrile high-speed homogenizations and extract 1 min, with centrifugal 5 min of 8000 r/min, solution changes in the clean eggplant-shape bottle, change in the residue extraction of ocean eddies 5 min on the vortex vortex mixer, repeated centrifugation step over to after cleaning the homogenate cutter head with 20 mL acetonitriles in addition, merge solution in eggplant-shape bottle, 40
oThe C water-bath is evaporated to that liquid is less than 2 mL in the bottle, takes out, and is washed till for three times in the 10 mL scale test tubes with 6mL moisture, and is settled to scale;
Tealeaves: the sample after taking by weighing 1.0 g pulverize in 100 mL polypropylene centrifuge tubes adds and extracts by above-mentioned steps behind the 5 mL water soaking 30min and hold again stand-by;
Sunflower seeds, rice: in 100 mL polypropylene centrifuge tubes, take by weighing the sample after 5.0 g pulverize, add 20 mL acetonitrile extraction of ocean eddies, 5 min, solution filter is to clean eggplant-shape bottle, residue repeats to extract once with 20 mL acetonitriles, and solution is incorporated in the above-mentioned eggplant-shape bottle, and 40
oIt is closely dried that the C water-bath is evaporated in the bottle liquid, takes out, and is washed till for three times in the 10 mL scale test tubes with 6mL moisture, and is settled to scale;
Meat, liver, egg, peeled shrimp: in 100 mL polypropylene centrifuge tubes, take by weighing the sample after 5.0 g pulverize, add 20 mL acetonitrile high-speed homogenizations and extract 1 min, with 8000 r/min frozen centrifugations (18
oC) 5 min, solution filter are to clean eggplant-shape bottle, and other changes in the residue after cleaning the homogenate cutter head with 20 mL acetonitriles, extraction of ocean eddies 5 min on the vortex vortex mixer, and the repeated centrifugation step merges solution in eggplant-shape bottle, 40
oThe C water-bath is evaporated to that liquid is less than 2 mL in the bottle, takes out, and is washed till for three times in the 10 mL scale test tubes with 6mL moisture, and is settled to scale;
Milk: in 100 mL polypropylene centrifuge tubes, take by weighing the sample of 2.0 g, add 20 mL acetonitrile extraction of ocean eddies 5min, with 8000 r/min frozen centrifugations (18
oC) 5 min, solution filter is to clean eggplant-shape bottle, in 40
oThe C water-bath is evaporated to that liquid is less than 2 mL in the bottle, takes out, and is washed till for three times in the 10 mL scale test tubes with 6mL moisture, and is settled to scale.
2. molecular engram solid phase extraction purifies:
The use operation steps of triazine phytocide molecular imprinting solid phase extracting material is as follows:
A, taking by weighing 50 mg triazine phytocide molecular imprinting solid phase extracting materials (MIPs), to join empty capacity be in band extraction plate (aperture 20 μ m) the polypropylene solid-phase extraction column of 10 mL, then add aperture 20 μ m sieve plates at filler, compress with the sieve plate insertion tool;
B, use 1 mL methyl alcohol successively, the pre-drip washing solid-phase extraction column of 1 mL water notes preventing filling surface dry in this process;
C, with in the aforesaid cabbages leaves extracting solution impouring post, coutroi velocity is no more than 0.5 mL/min, after solution all flows out, with 1 mL water wash, discards whole effluent liquid, then drain cylinder, and state 30 min is drained in maintenance continuously;
D, with 3 * 1 mL methylene dichloride (CH
2Cl
2) the drip washing cylinder, be evacuated to cylinder with vacuum pump between each drip washing closely dried, with 3 * 1 mL methanol-eluted fractions, coutroi velocity is no more than 0.2 mL/min again, is evacuated to cylinder with vacuum pump between each wash-out closely dried;
E, with elutriant on Nitrogen evaporator 40
oThe C nitrogen flushing is concentrated into dried, holds again with 1 mL moving phase, and crosses 0.22 μ m organic system syringe needle filter membrane, and clear liquid is measured for liquid chromatography-tandem mass spectrometry.
3. instrument detects
Take Agilent 1200-API 4000 Q-trap type Liquid Chromatography-Tandem Mass Spectrometry systems (U.S. Applied Biosystem company) as example:
Liquid-phase condition: chromatographic column is Waters Xterra MS C
18, specification is 2.1 mm*100 mm, 3.5 μ m.Mobile phase A is acetonitrile; B is 0.05% aqueous formic acid (V/V); Gradient elution: 0～5 min, 20～70% A; 5.1～10 min, 70%A; 10.1～20 min, 20% A.Flow velocity: 200 μ L/min; Sample size: 10 μ L; Column temperature: 40 ℃;
Mass spectrum condition: electron spray(ES) voltage (IS): 5500 V; Atomization gas pressure (GS1): 60 psi; Gas curtain atmospheric pressure (CUR): 20 psi; Assisted gas flow velocity (GS2): 60 psi; Ion source temperature (TEM): 450
oC; Collision gas (CAD): medium tenacity (Medium); Residence time: 200 ms; Holotype MRM monitoring.
The qualitative, quantitative ion pair of table 17 triazine herbicide and DP, CE value
Claims (1)
1. the preparation method of triazine phytocide molecular imprinting solid phase extracting material is characterized in that comprising following operation steps:
1.1 the preparation of suspending phase I
Get melamine-butyl, α-methacrylic acid (MAA), ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) and Diisopropyl azodicarboxylate (AIBN), be dissolved in trichloromethane, mix, get suspending phase I;
Wherein melamine-butyl is imprinted templates, and α-methacrylic acid (MAA) is function monomer, and ethylene glycol bisthioglycolate (methacrylic acid) ester (EDGMA) is linking agent, and Diisopropyl azodicarboxylate (AIBN) is initiator, and trichloromethane is solvent;
The molar ratio of imprinted templates and function monomer is 1:3, and the molar ratio of linking agent and function monomer is 4:1～6:1, and the molar ratio of initiator and function monomer is 1:4～1:6, and the volumetric usage of solvent and linking agent molar ratio are 1:2～1:4;
1.2 the preparation of suspending phase II
It is in 80～100 ℃ the hot water that polyvinyl alcohol 1788 is dissolved in temperature, then is cooled to room temperature, gets suspending phase II; Wherein polyvinyl alcohol 1788 is protective material, and water is disperse phase; The consumption of polyvinyl alcohol 1788 is that every 50mL water adds 1g, and the ratio of the volumetric usage of water and suspending phase I volume is 4:1～6:1;
1.3 the preparation of mixture
With slowly impouring suspending phase of suspending phase I II, and stir, get mixing solutions; Under 50 ° of C water-bath magnetic agitation conditions, with 80～120 turn/rotating speed of min mixes solution reaction 6h, continues stirring reaction 15h under room temperature again, adopts the method collecting precipitation of filter paper filtering at the solid particulate matter of solution bottom;
1.4 remove imprinted templates and vacuum-drying
Solid particulate matter take methyl alcohol as solvent, is removed melamine-butyl and unreacted reactant in 24 hours through the Soxhlet extraction, again through 40 ° of C vacuum-dryings, get the triazine phytocide molecular imprinting solid phase extracting material of white powder;
The consumption that each Soxhlet is extracted methyl alcohol is 200mL.
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| CN103433009B (en) * | 2013-08-30 | 2015-11-18 | 江苏大学 | A kind of prometryn molecularly imprinted solid phase extraction column and using method |
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| CN106188397B (en) * | 2016-08-10 | 2018-05-25 | 昆明理工大学 | A kind of preparation method of triazine herbicide molecularly imprinted polymer |
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| CN108593823B (en) * | 2018-04-04 | 2020-10-09 | 桂林理工大学 | Method for separating and enriching triazine pesticides in large-volume water sample |
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