CN105032381A - Compound molecularly imprinted solid phase extraction column as well as preparation method and application thereof - Google Patents
Compound molecularly imprinted solid phase extraction column as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a preparation method of a compound molecularly imprinted solid phase extraction column. The preparation method comprises the following steps: a, preparing a molecularly imprinted polymer A by taking a triazine pesticide as a template molecule; B, preparing a molecularly imprinted polymer B by taking a sulfonylurea pesticide as a template molecule; and C, preparing the compound molecularly imprinted solid phase extraction column. The compound molecularly imprinted solid phase extraction column is capable of selectively enriching and extracting the triazine pesticide and sulfonylurea pesticide in a sample at the same time, is simple and convenient to use and good in enrichment and extraction effect, can be repeatedly used for at least twenty times, has the advantages of economical efficiency, environmental friendliness, low cost and the like, and is quite applicable to enriching and extracting the triazine pesticide and the sulfonylurea pesticide at the same time. The compound molecularly imprinted solid phase extraction column is accurate and reliable in detection result when used for detecting based on an HPLO (High Performance Liquid Chromatography)-tandem mass spectrometry method.
Description
Technical field
The present invention relates to compound molecule trace solid-phase extraction column and preparation method thereof and application.
Background technology
Triazines and sulfonylurea herbicide are class herbicides with hexa-member heterocycle structure, since release, just have worldwide widely used to prevent farmland weed to grow, especially for cereal crops as highy potent herbicide.Triazine herbicide is widely used all over the world, but the residual meeting of this type of herbicide in crops brings certain harm to human body, and its safety issue residual in crops has caused increasing concern.
Molecularly imprinted polymer (MIPs) optionally can adsorb the target compound in complex matrices, there is the stability of certain mechanical strength and acid and alkali-resistance heat simultaneously, and long service life, affinity are high, selective good, be a kind of SPE material of function admirable, molecularly imprinted solid phase extraction column can be made into and use.Compared with traditional SPE, molecular engram solid phase extraction is with low cost, simple to operate, effectively can remove sample substrate interference simultaneously, improve the accuracy of analysis, be more suitable for trace analysis.
At present, disclosed molecularly imprinted solid phase extraction column is mainly unimolecule trace solid-phase extraction column, as: the hydrophily Sulfa drugs molecule trace solid-phase extraction column of Chinese CN104001486A, the aminoglycoside medicaments molecularly imprinted solid phase extraction column of Chinese patent CN102974327A, the Coumarins rat poison molecularly imprinted solid phase extraction column of Chinese patent CN102631902A, this unimolecule trace solid-phase extraction column can only extract a certain class medicine, then carry out detection to analyze, and the medicine different to two classes can not be realized extract simultaneously, detect; In prior art, there are no utilizing compound molecule trace solid-phase extraction column simultaneously to the relevant report that medicines more than two classes or two classes extracts, detects.
Therefore, need to invent a kind of novel molecularly imprinted solid phase extraction column, it can extract medicines more than two classes or two classes, detect simultaneously.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of compound molecule trace solid-phase extraction column.
The preparation method of a kind of compound molecule trace solid-phase extraction column provided by the invention, it comprises the following steps:
A, be that template molecule prepares molecularly imprinted polymer A with triazine:
Triazine dissolves in organic solvent, adds function monomer A, and concussion, adds crosslinking agent and initator, ultrasonic, fills N
2deoxygenation, after sealing, reaction, obtains polymer A; Polymer A is pulverized, crosses 200 mesh sieves, wash away template molecule, obtain molecularly imprinted polymer A;
The mol ratio of triazine, function monomer A, crosslinking agent, initator is 1:3 ~ 5:3 ~ 5:0.5 ~ 0.8; Triazine is 1:8 ~ 12mmol/mL with the molal volume ratio of organic solvent;
B, be that template molecule prepares molecularly imprinted polymer B with sulfonylurea pesticide:
Sulfonylurea pesticide dissolves in organic solvent, adds function monomer B, and concussion, adds crosslinking agent and initator, ultrasonic, fills N
2deoxygenation, after sealing, reaction, obtains polymer B; Wash away template molecule, obtain molecularly imprinted polymer B;
The mol ratio of sulfonylurea pesticide, function monomer B, crosslinking agent, initator is 1:3 ~ 5:3 ~ 5:0.5 ~ 0.8; Sulfonylurea pesticide is 1:150 ~ 170mmol/mL with the molal volume ratio of organic solvent;
C, prepare compound molecule trace solid-phase extraction column:
Using molecularly imprinted polymer A and molecularly imprinted polymer B as filler, be filled into bottom and be equipped with in the solid-phase extraction column void column of sieve plate, upper end adds upper sieve plate, compresses, obtains compound molecule trace solid-phase extraction column; The weight ratio of molecularly imprinted polymer A and molecularly imprinted polymer B is 1:4 ~ 6.
Further,
In step a, triazine is selected from that ametryn, prometryn, cyanazine, cyprazine, Simanex, dipropetryn, methoxy third are clean, symetryne, Gesadrual, atraton, Garagard, propazine, de-isopropyl-Atrazine, de-ethyl-Atrazine, Te Dingtong, prometon, in Atrazine any one or multiple; Function monomer A is methacrylic acid; Crosslinking agent is trimethyl propane trimethyl acrylic ester; Initator is azodiisobutyronitrile; Organic solvent is selected from acetonitrile or carrene;
In step b, sulfonylurea pesticide is selected from that chlorine sulphur is grand, in monosulfmeturon, thifensulfuronmethyl any one or multiple; Function monomer B is diethylaminoethyl methacrylate; Crosslinking agent is trimethyl propane trimethyl acrylic ester; Initator is azodiisobutyronitrile; Organic solvent is selected from acetonitrile or carrene;
In step c, molecularly imprinted polymer A is selected from ametryn molecularly imprinted polymer, prometryn molecularly imprinted polymer, cyanazine molecularly imprinted polymer, cyprazine molecularly imprinted polymer, Simanex molecularly imprinted polymer, dipropetryn molecularly imprinted polymer, the clean molecularly imprinted polymer of methoxy third, symetryne molecularly imprinted polymer, Gesadrual molecularly imprinted polymer, atraton molecularly imprinted polymer, Garagard molecularly imprinted polymer, propazine molecularly imprinted polymer, de-isopropyl-Atrazine molecularly imprinted polymer, de-ethyl-Atrazine molecularly imprinted polymer, Te Dingtong molecularly imprinted polymer, prometon molecularly imprinted polymer, in Atrazine molecularly imprinted polymer any one or multiple, molecularly imprinted polymer B be selected from the grand molecularly imprinted polymer of chlorine sulphur, monosulfmeturon molecularly imprinted polymer, thifensulfuronmethyl molecularly imprinted polymer any one or multiple.
Further, molecularly imprinted polymer A comprises atrazine molecularly imprinted polymer and prometryn molecularly imprinted polymer; Molecularly imprinted polymer B is the grand molecularly imprinted polymer of chlorine sulphur.
Further,
In step a, the mol ratio of triazine, function monomer A, crosslinking agent, initator is 1:4:4:0.6; Triazine is 1:10mmol/mL with the molal volume ratio of organic solvent;
In step b, the mol ratio of sulfonylurea pesticide, function monomer B, crosslinking agent, initator is 1:4:4:0.6; Sulfonylurea pesticide is 1:160mmol/mL with the molal volume ratio of organic solvent;
In step c, the weight ratio of molecularly imprinted polymer A and molecularly imprinted polymer B is 1:5.
Further, in molecularly imprinted polymer A, the weight ratio of atrazine molecularly imprinted polymer and prometryn molecularly imprinted polymer is 5:3.
Further,
In step a, the concussion time is 30min; Reaction temperature is 50 DEG C ~ 70 DEG C; Reaction time is 20h ~ 28h; In Soxhlet extraction device, template molecule is washed away with 10% acetic acid methanol solution;
In step b, the concussion time is 30min; Reaction temperature is 50 DEG C ~ 70 DEG C; Reaction time is 20h ~ 28h; In Soxhlet extraction device, template molecule is washed away with 10% acetic acid methanol solution.
Further,
In step a, the concussion time is 30min; Reaction temperature is 60 DEG C; Reaction time is 24h;
In step b, the concussion time is 30min; Reaction temperature is 60 DEG C; Reaction time is 24h.
Present invention also offers compound molecule trace solid-phase extraction column prepared by said method.
A kind of compound molecule trace solid-phase extraction column provided by the invention, it is the solid-phase extraction column prepared by said method.
Present invention also offers the sample-pretreating method of Detecting Pesticide, it comprises the following steps:
I, get the sample of homogeneous preparation, mix with acetonitrile or the saturated acetonitrile solution of n-hexane, mechanical shaking extraction, centrifugal, repeat extraction twice, merge supernatant, be acetonitrile extract;
Ii, get acetonitrile extract, be concentrated into dry, redissolve with acetonitrile, obtain sample liquid;
Iii, sampling liquid cross the compound molecule trace solid-phase extraction column of claim 8, and coutroi velocity is 0.3mL/min, drains off; With water wash, drain off; Use methyl alcohol, acetate-methanol wash-out successively, collection wash-out liquid nitrogen blows and is concentrated near doing, and adds acetonitrile and dissolves, and mixes, obtain need testing solution with 0.1% aqueous formic acid dilution.
Further,
In step I, described sample is plant-derived food; The w/v of sample and acetonitrile is 2:15g/ml; The time of mechanical shaking extraction is 10min; Centrifugal rotating speed is 4000r/min, and the centrifugal time is 5min;
In step I i, parallel quantitative concentrating instrument is adopted to be concentrated into dry; Temperature time concentrated is 50 DEG C;
In step I ii, the volume ratio of sample liquid, pouring scouring water, methyl alcohol, acetate-methanol, acetonitrile, 0.1% aqueous formic acid is 1:1:2:6:1:1; In acetate-methanol, the volume ratio of acetic acid and methyl alcohol is 1:9.
Present invention also offers the method that high performance liquid chromatography-tandem mass method detects sample Pesticide Residues, it comprises the following steps:
1., according to sample-pretreating method according to claim 9, need testing solution is obtained;
2., high performance liquid chromatography-tandem mass method is adopted to detect need testing solution:
High-efficient liquid phase chromatogram condition:
Chromatographic column: AgilentZORBAXRX-C8 post;
Mobile phase A: 0.1% formic acid-aqueous solution, Mobile phase B: acetonitrile;
Gradient elution program is as follows:
| Time | Mobile phase A | Mobile phase B |
| 0min～5min | 10％→60％ | 90％→40％ |
| 5min～11min | 60％ | 40％ |
| 11min～11.1min | 60％→10％ | 40％→90％ |
| 11.1min～13min | 10％ | 90％ |
Tandem mass spectrum condition: electric spray ion source.
Further,
Step 2. in, high-efficient liquid phase chromatogram condition: column temperature is 30 DEG C, flow velocity is 200 μ L/min, and sample size is 3 μ L; Tandem mass spectrum condition: ion source temperature is 450 DEG C, cation scans, multiple-reaction monitoring mode detection, and atomization gas pressure is 30psi, and heating assisted gas pressure is 30psi, and gas curtain atmospheric pressure is 35psi, and spray voltage is 5500V.
Compound molecule trace solid-phase extraction column of the present invention, can triazine simultaneously in selective enrichment, extraction sample and sulfonylurea pesticide, easy to use, enrichment, extraction effective, and can at least reuse 20 times, there is economic environmental protection, low cost and other advantages, be applicable to very much carrying out enrichment, extraction to triazine and sulfonylurea pesticide simultaneously, when the high performance liquid chromatography-tandem mass method that is used for compound molecule trace solid-phase extraction column of the present invention detects, testing result accurately and reliably.
Obviously, according to foregoing of the present invention, according to ordinary technical knowledge and the customary means of this area, not departing under the present invention's above-mentioned basic fundamental thought prerequisite, the amendment of other various ways, replacement or change can also be made.
The detailed description of the invention of form by the following examples, is described in further detail foregoing of the present invention again.But this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example.All technology realized based on foregoing of the present invention all belong to scope of the present invention.
Accompanying drawing explanation
Fig. 1 is the SEM figure of atrazine molecularly imprinted polymer of the present invention;
Fig. 2 is the SEM figure of prometryn molecularly imprinted polymer of the present invention;
Fig. 3 is the SEM figure of the grand molecularly imprinted polymer of chlorine sulphur of the present invention;
Fig. 4 is total ion current (totalioncurrent, the TIC) chromatogram of 20 kinds of agricultural chemicals in the corn sample of mark-on level 10 μ g/kg;
Fig. 5 is the grand mass spectrogram of chlorine sulphur.
Fig. 6 is the mass spectrogram of de-isopropyl-Atrazine;
Fig. 7 is the TIC of typical positive sample.
Detailed description of the invention
The raw material used in the specific embodiment of the invention, equipment are known product, obtain by buying commercially available prod.
Embodiment 1
1.1, instrument and reagent
AB5500 high performance liquid chromatography-triple quadrupole rods tandem mass spectrometry instrument, is furnished with electric spray ion source (ESI) and Analyst1.6.2 work station (American AB SCIEX company); Parallel quantitative concentrating instrument, B-740 recirculated cooling system, V-700/701 vavuum pump (BUCHI company of Switzerland); Nitrogen evaporator (Biotage company of Sweden); Supercentrifuge (Beckman company of the U.S.);
Acetonitrile, methyl alcohol (chromatographically pure, Fisher company), n-hexane (chromatographically pure, Honeywell company), formic acid (chromatographically pure, German CNW company); Acetic acid (chromatographically pure, the general scientific instrument Co., Ltd of Town in Shanghai);
20 kinds of standard sample of pesticide: ametryn (ametryn), prometryn (prometryn), cyanazine (cyanazine), cyprazine (cyprazine), Simanex (simazine), dipropetryn (dipropetryn), methoxy third clean (methoprotryne), symetryne (simetryn), Gesadrual (simeton), atraton (atraton), Garagard (terbuthylazine), propazine (propazine), de-isopropyl-Atrazine (atrazine-desisopropyl), de-ethyl-Atrazine (atrazine-desethyl), Te Dingtong (terbumeton), prometon (prometon), Atrazine (atrazine), chlorine sulphur grand (chlorsulfuron), monosulfmeturon (monosulfuron), (purity is all greater than 97% to thifensulfuronmethyl (thifensulfuron-methyl), be purchased from German Dr.Ehrenstorfer company),
Experimental water is Milli-Q water (ultra-pure water of distilled water after Millipore, Bedford, MA, USA produce water purification machine process).
1.2, the preparation of standard liquid
Single standard storing solution: the standard sample of pesticide accurately taking 5mg (being accurate to 0.01mg) respectively, be placed in 10mL volumetric flask, dissolve with methyl alcohol and dilute constant volume, being made into the standard reserving solution that mass concentration is 500 μ g/mL, at-18 DEG C, lucifuge stores.
Mixed standard solution: accurately draw each pesticide standard stock solution of 1mL in 50mL volumetric flask, use dilution in acetonitrile constant volume, be mixed with the hybrid standard working solution that each pesticide composition is 10 μ g/mL, keep in Dark Place under 4 DEG C of conditions.During use with methanol dilution to finite concentration, carry out the detection of high performance liquid chromatography-tandem mass method.
1.3, the preparation of compound molecule trace SPE (CMISPE) post
The preparation of a, triazine molecularly imprinted polymer:
In the acetonitrile that prometryn (0.2mmol) or atrazine (0.2mmol) are dissolved in certain volume or carrene (2mL), add function monomer MAA (methacrylic acid wherein, 0.8mmol), concussion 30min, then a certain amount of crosslinking agent TRIM (trimethyl propane trimethyl acrylic ester, 0.8mmol) and initiator A IBN (azodiisobutyronitrile, 20mg) is added, ultrasonic 5min, fills N
2deoxygenation 10min, put into water-bath after sealing and react 24h at 60 DEG C, reacted rear taking-up polymer, after grinding, cross 200 mesh sieves, in Soxhlet extraction device, wash away template with 10% acetic acid methanol solution, obtain atrazine molecularly imprinted polymer, prometryn molecularly imprinted polymer; Its ESEM testing result is shown in Fig. 1 and Fig. 2 respectively.
The preparation of b, sulfonylurea pesticide molecularly imprinted polymer:
Chlorine sulphur grand (0.2mmol) is dissolved in the acetonitrile (32mL) of certain volume, add function monomer DEMEA (diethylaminoethyl methacrylate, 0.8mmol), concussion 30min, then a certain amount of crosslinking agent TRIM (0.8mmol) and initiator A IBN20mg is added, ultrasonic, fill N
2deoxygenation, puts into water-bath and react 24h at 60 DEG C after sealing, reacted rear taking-up polymer, washed away template with 10% acetic acid methanol in Soxhlet extraction device, obtains the grand molecularly imprinted polymer of chlorine sulphur; Its ESEM testing result is shown in Fig. 3.
The preparation of c, compound molecule trace solid-phase extraction column (CMISPE post):
Take the grand molecularly imprinted polymer mix suspending of 50mg atrazine molecularly imprinted polymer, 30mg prometryn molecularly imprinted polymer and 400mg chlorine sulphur in acetonitrile, being filled into bottom is equipped with in the solid-phase extraction column void column of sieve plate, upper end adds sieve plate again, compresses and makes CMISPE post.
1.4, the sample-pretreating method of Detecting Pesticide
Extract:
The sample 2.0g accurately taking homogeneous preparation is placed in 50mL centrifuge tube, add 15mL acetonitrile, vortex mixing 1min, the centrifugal 5min of mechanical shaking extraction 10min, 4000r/min, repeats extraction twice again, merge supernatant (for the sample that the fat contents such as corn are higher, add the saturated acetonitrile solution of 20mL n-hexane (take off layer solution after the mixing of n-hexane, acetonitrile and be the saturated acetonitrile solution of n-hexane) mechanical shaking extraction, discard n-hexane layer), obtain acetonitrile extract.Acetonitrile extract parallel quantitative concentrating instrument (temperature is adjusted to 50 DEG C) is concentrated into dry, redissolves, get wherein 1mL sample liquid with 2mL acetonitrile, to be clean.
SPE:
With 3mL acetonitrile activation CMISPE post, acetonitrile is not made to drain off; Join in solid-phase extraction column by 1mL sample liquid, coutroi velocity is about 0.3mL/min, drains off; Use 1mL water wash, drain off; Use 2 × 1mL methyl alcohol and 6 × 1mL acid alcohol solution (acetic acid: methyl alcohol (V/V)=1:9) wash-out successively, collection wash-out liquid nitrogen blows and is concentrated near doing, add 1.0mL acetonitrile to dissolve, mixing is diluted to 2.0mL with mobile phase 0.1% aqueous formic acid 1:1, as need testing solution, upper machine analysis.
Regeneration:
With 4mL acid alcohol solution (acetic acid: methyl alcohol (V/V)=1:9) drip washing CMISPE post more than 3 times, after washing out to driftlessness thing, pillar can reuse more than 20 times.
1.5, high performance liquid chromatography-tandem mass method is adopted to detect need testing solution:
High-efficient liquid phase chromatogram condition:
Chromatographic column: AgilentZORBAXRX-C8 post (150mm × 2.1mmi.d, 5 μm) (i.d represents the internal diameter of chromatographic column); Mobile phase A: 0.1% formic acid-aqueous solution, Mobile phase B: acetonitrile; Gradient elution program: in table 1; Flow velocity: 200 μ L/min; Column temperature: 30 DEG C; Sample size: 3 μ L.
Table 1, gradient elution program of the present invention
| Time | Mobile phase A | Mobile phase B |
| 0min～5min | 10％→60％ | 90％→40％ |
| 5min～11min | 60％ | 40％ |
| 11min～11.1min | 60％→10％ | 40％→90％ |
| 11.1min～13min | 10％ | 90％ |
Tandem mass spectrum condition:
Electric spray ion source (ESI); Ion source temperature: 450 DEG C; Cation scans; Multiple-reaction monitoring (MRM) mode detection; Atomization gas (Gas1) pressure: 30psi; Heating assisted gas (Gas2) pressure: 30psi; Gas curtain gas (CUR) pressure: 35psi; Spray voltage (IS): 5500V; All the other mass spectrometry parameters are in table 2 (get the mixed standard solution of 20ng/mL, adopt the mode of pin pump sample introduction, determine the mass spectrometry parameters of each compound).
The Mass Spectrometer Method condition of table 2,20 kinds of herbicides and retention time
Note: * is quota ion
Compound molecule trace solid-phase extraction column of the present invention, has the following advantages:
(1), the present invention first by triazine herbicide molecularly imprinted polymer and sulfonylurea weedicide molecularly imprinted polymer mixing be filled in solid-phase extraction column, realize the Sync enrichment of 17 kinds of triazines and 3 kinds of sulfonylurea pesticides;
(2), compound molecule trace solid-phase extraction column of the present invention, using two kinds of molecularly imprinted polymer mixing as filler, current universal solid-phase extraction column poor selectivity, problem that matrix effect is large can be solved, the problem that current molecular trace solid-phase extraction column bound species is few can also be solved, can selective cleaning, trace triazine in enriched sample and sulfonylurea pesticide, be 62.7% ~ 117.4% to the rate of recovery of two class herbicides totally 20 kinds of materials, meet the requirement that trace residue detects;
(3), compared with universal solid-phase extraction column, compound molecule trace solid phase column of the present invention, pretreatment process is simple, step is few, enrichment purification effective, organic reagent use amount is few, economic environmental protection, and at least can reuse 20 times after regeneration.
In order to beneficial effect of the present invention is described, the invention provides following test example:
The screening of test example 1 Extraction solvent
Relatively acetonitrile, methyl alcohol, acetone, ethyl acetate 4 kinds of solvents add the extraction efficiency that concentration is 10 μ g/kg in corn sample.
Found that, using methyl alcohol as Extraction solvent, the rate of recovery of its correspondence is 43% ~ 78%; Be 45% ~ 88% during using acetone as Extraction solvent; Be 58.4% ~ 109.8% during using ethyl acetate as Extraction solvent; And pigment content is higher in acetone, methanol extract liquid, the boiling point of ethyl acetate is higher, concentrated consuming time too of a specified duration, is all not suitable for use in Extraction solvent.
When the present invention adopts acetonitrile as Extraction solvent, extraction efficiency is 79% ~ 116%, and extraction effect is also even more ideal.
The screening of test example 2 purification method
Purification method comprises liquid-liquid extraction and molecular engram solid phase extraction.The sample such as corn, soybean fat content is more, and extract can be caused muddy, and n-hexane is usually used in extracting degreasing, therefore for this type of sample, after extracting, select n-hexane to carry out liquid-liquid extraction, most of oil lipid chaff interference can be removed, then thorough to purify after post.Considering that n-hexane has certain solubility in acetonitrile, for ensureing the rate of recovery, thus selecting the saturated n-hexane of acetonitrile to purify.
The screening test of test example 3 SPE
Acetonitrile, methyl alcohol, acid alcohol (acetic acid: methyl alcohol (V/V)=1:9) and water 4 kinds of different adsorption solvent are investigated to the impact of absorption property, adopt above-mentioned 4 kinds of different eluent solvent compound molecule imprinting solid phase extraction small columns after loading, detect the rate of recovery situation of 20 kinds of pesticides in leacheate.
Result shows, when water and acetonitrile are as adsorbing medium, better to most of herbicide absorption property, in leacheate, almost driftlessness detects thing, but can cause the partial loss of monosulfmeturon and thifensulfuronmethyl in Acetonitrile, thus this research adopts water as eluent.Because the hydrogen bond action constant of methyl alcohol is much larger than acetonitrile, so the absorption property of imprinted polymer in METHANOL MEDIUM is minimum, thus have selected methyl alcohol as eluting solvent, adding acetic acid can destroy Hyarogen-bonding further, makes target compound wash-out complete.Test compares eluting power when methyl alcohol, acid alcohol and both conbined usage, found that the elute effect using methyl alcohol, acid alcohol can reach best successively, and investigate the impact of elution volume on the rate of recovery, result shows, as use methyl alcohol 2mL, during acid alcohol 6mL, target compound wash-out is complete, can ensure enough rate of recovery.
The screening of test example 4 high performance liquid chromatography-tandem mass method testing conditions
Sulfonylurea herbicide is faintly acid, and mobile phase through acid or glacial acetic acid acidifying, and adds the ionization that 0.1% formic acid is more conducive to compound in water, can improve compound peak shape.Select herein with acetonitrile-0.1% formic acid for mobile phase, the separation completing 20 kinds of agricultural chemicals in 13min detects, and peak shape is sharp-pointed, separating degree is better.
In the corn sample of mark-on level 10 μ g/kg, total ion current (totalioncurrent, the TIC) chromatogram of 20 kinds of agricultural chemicals, is shown in Fig. 4.
The mass spectrogram that chlorine sulphur is grand, is shown in Fig. 5; The mass spectrogram of de-isopropyl-Atrazine, is shown in Fig. 6.
Test example 5, the range of linearity and quantitative limit
Determining, under optimum analysis condition, the mixed standard solution of variable concentrations to be carried out HPLC-MS/MS analysis.Be abscissa respectively with testing concentration, peak area is that ordinate carries out linear regression.
Result shows: 20 kinds of herbicides are good in 0.5 ~ 20ng/mL concentration range internal linear relation, and coefficient correlation is 0.9942 ~ 0.9999, can meet the requirement of quantitative analysis.Be limited to 0.057 ~ 0.63 μ g/kg with detecting of S/N >=3 defining method, be quantitatively limited to 0.19 ~ 2.1 μ g/kg with S/N >=10 defining method.The linear relationship of 20 kinds of triazines and sulfonylurea herbicide, the range of linearity and quantitative limit are in table 3.
The coefficient correlation of 20 kinds of herbicides, quantitative limit and the rate of recovery (n=6) in table 3, corn
In equation of linear regression, e4 represents 4 powers of 10, and other are similar.
The veracity and precision of test example 6 method
In blank corn sample, add mixed standard solution makes final concentration be 5 μ g/kg, 10 μ g/kg, 20 μ g/kg, carries out the pre-treatment of sample, each concentration level replication 6 times according to 1.4; Average recovery rate and precision the results are shown in Table 2.
Result shows, the average recovery rate of each compound is between 62.7% ~ 117.4%, and relative standard deviation (RSD) is 1.7% ~ 13.9%, meets the technical requirement that food Physico-chemical tests method confirms.
10 kinds of different samples are selected according to GB GB2763-2014 " Pesticide MRL " and Hong Kong " in food the remaining regulations of pesticide ", application this method detects it, the results are shown in Table 4, in corn, glutinous corn, brown rice, pineapple, lemon 5 kinds of samples, detect 6 kinds of residues of pesticides altogether respectively.
The chromatogram of typical positive sample, is shown in Fig. 7.
Table 4, do not plant the testing result of sample
| Sample | Thifensulfuronmethyl | De-isopropyl-Atrazine | De-ethyl-Atrazine | Ametryn | Cyprazine | Chlorine sulphur is grand |
| Corn | ND | ND | ND | 0.6205 | ND | 3.0828 |
| Glutinous corn | 2.8964 | 3.6423 | ND | ND | ND | ND |
| Wheat | ND | ND | ND | ND | ND | ND |
| Brown rice | ND | ND | ND | 0.2487 | ND | ND |
| Pineapple | ND | ND | ND | 0.4899 | 0.2993 | ND |
| Strawberry | ND | ND | ND | ND | ND | ND |
| Sugarcane | ND | ND | ND | ND | ND | ND |
| Lemon | ND | ND | 2.0021 | ND | ND | ND |
| Apple | ND | ND | ND | ND | ND | ND |
| Car li son | ND | ND | ND | ND | ND | ND |
ND: do not detect (notdetected).
In sum, compound molecule trace solid-phase extraction column of the present invention, can triazine simultaneously in selective enrichment, extraction sample and sulfonylurea pesticide, easy to use, enrichment, extraction effective, and can at least reuse 20 times, there is economic environmental protection, low cost and other advantages, be applicable to very much carrying out enrichment, extraction to triazine and sulfonylurea pesticide simultaneously, when the high performance liquid chromatography-tandem mass method that is used for compound molecule trace solid-phase extraction column of the present invention detects, testing result accurately and reliably.
Claims (10)
1. a preparation method for compound molecule trace solid-phase extraction column, is characterized in that: it comprises the following steps:
A, be that template molecule prepares molecularly imprinted polymer A with triazine:
Triazine dissolves in organic solvent, adds function monomer A, and concussion, adds crosslinking agent and initator, ultrasonic, fills N
2deoxygenation, after sealing, reaction, obtains polymer A; Polymer A is pulverized, crosses 200 mesh sieves, wash away template molecule, obtain molecularly imprinted polymer A;
The mol ratio of triazine, function monomer A, crosslinking agent, initator is 1:3 ~ 5:3 ~ 5:0.5 ~ 0.8; Triazine is 1:8 ~ 12mmol/mL with the molal volume ratio of organic solvent;
B, be that template molecule prepares molecularly imprinted polymer B with sulfonylurea pesticide:
Sulfonylurea pesticide dissolves in organic solvent, adds function monomer B, and concussion, adds crosslinking agent and initator, ultrasonic, fills N
2deoxygenation, after sealing, reaction, obtains polymer B; Wash away template molecule, obtain molecularly imprinted polymer B;
The mol ratio of sulfonylurea pesticide, function monomer B, crosslinking agent, initator is 1:3 ~ 5:3 ~ 5:0.5 ~ 0.8; Sulfonylurea pesticide is 1:150 ~ 170mmol/mL with the molal volume ratio of organic solvent;
C, prepare compound molecule trace solid-phase extraction column:
Using molecularly imprinted polymer A and molecularly imprinted polymer B as filler, be filled into bottom and be equipped with in the solid-phase extraction column void column of sieve plate, upper end adds upper sieve plate, compresses, obtains compound molecule trace solid-phase extraction column; The weight ratio of molecularly imprinted polymer A and molecularly imprinted polymer B is 1:4 ~ 6.
2. preparation method according to claim 1, is characterized in that:
In step a, triazine is selected from that ametryn, prometryn, cyanazine, cyprazine, Simanex, dipropetryn, methoxy third are clean, symetryne, Gesadrual, atraton, Garagard, propazine, de-isopropyl-Atrazine, de-ethyl-Atrazine, Te Dingtong, prometon, in Atrazine any one or multiple; Function monomer A is methacrylic acid; Crosslinking agent is trimethyl propane trimethyl acrylic ester; Initator is azodiisobutyronitrile; Organic solvent is selected from acetonitrile or carrene;
In step b, sulfonylurea pesticide is selected from that chlorine sulphur is grand, in monosulfmeturon, thifensulfuronmethyl any one or multiple; Function monomer B is diethylaminoethyl methacrylate; Crosslinking agent is trimethyl propane trimethyl acrylic ester; Initator is azodiisobutyronitrile; Organic solvent is selected from acetonitrile or carrene;
In step c, molecularly imprinted polymer A is selected from ametryn molecularly imprinted polymer, prometryn molecularly imprinted polymer, cyanazine molecularly imprinted polymer, cyprazine molecularly imprinted polymer, Simanex molecularly imprinted polymer, dipropetryn molecularly imprinted polymer, the clean molecularly imprinted polymer of methoxy third, symetryne molecularly imprinted polymer, Gesadrual molecularly imprinted polymer, atraton molecularly imprinted polymer, Garagard molecularly imprinted polymer, propazine molecularly imprinted polymer, de-isopropyl-Atrazine molecularly imprinted polymer, de-ethyl-Atrazine molecularly imprinted polymer, Te Dingtong molecularly imprinted polymer, prometon molecularly imprinted polymer, in Atrazine molecularly imprinted polymer any one or multiple, molecularly imprinted polymer B be selected from the grand molecularly imprinted polymer of chlorine sulphur, monosulfmeturon molecularly imprinted polymer, thifensulfuronmethyl molecularly imprinted polymer any one or multiple.
3. preparation method according to claim 2, is characterized in that: molecularly imprinted polymer A comprises atrazine molecularly imprinted polymer and prometryn molecularly imprinted polymer; Molecularly imprinted polymer B is the grand molecularly imprinted polymer of chlorine sulphur.
4. the preparation method according to claims 1 to 3 any one, is characterized in that:
In step a, the mol ratio of triazine, function monomer A, crosslinking agent, initator is 1:4:4:0.6; Triazine is 1:10mmol/mL with the molal volume ratio of organic solvent;
In step b, the mol ratio of sulfonylurea pesticide, function monomer B, crosslinking agent, initator is 1:4:4:0.6; Sulfonylurea pesticide is 1:160mmol/mL with the molal volume ratio of organic solvent;
In step c, the weight ratio of molecularly imprinted polymer A and molecularly imprinted polymer B is 1:5.
5. preparation method according to claim 4, is characterized in that: in molecularly imprinted polymer A, and the weight ratio of atrazine molecularly imprinted polymer and prometryn molecularly imprinted polymer is 5:3.
6. the preparation method according to claims 1 to 3 any one, is characterized in that:
In step a, the concussion time is 30min; Reaction temperature is 50 DEG C ~ 70 DEG C; Reaction time is 20h ~ 28h; In Soxhlet extraction device, template molecule is washed away with 10% acetic acid methanol solution;
In step b, the concussion time is 30min; Reaction temperature is 50 DEG C ~ 70 DEG C; Reaction time is 20h ~ 28h; In Soxhlet extraction device, template molecule is washed away with 10% acetic acid methanol solution.
7. preparation method according to claim 6, is characterized in that:
In step a, the concussion time is 30min; Reaction temperature is 60 DEG C; Reaction time is 24h;
In step b, the concussion time is 30min; Reaction temperature is 60 DEG C; Reaction time is 24h.
8. a compound molecule trace solid-phase extraction column, is characterized in that: it is the solid-phase extraction column prepared by the method for claim 1 ~ 7 any one.
9. the sample-pretreating method of Detecting Pesticide, is characterized in that: it comprises the following steps:
I, get the sample of homogeneous preparation, mix with acetonitrile or the saturated acetonitrile solution of n-hexane, mechanical shaking extraction, centrifugal, repeat extraction twice, merge supernatant, be acetonitrile extract;
Ii, get acetonitrile extract, be concentrated into dry, redissolve with acetonitrile, obtain sample liquid;
Iii, sampling liquid cross the compound molecule trace solid-phase extraction column of claim 8, and coutroi velocity is 0.3mL/min, drains off; With water wash, drain off; Use methyl alcohol, acetate-methanol wash-out successively, collection wash-out liquid nitrogen blows and is concentrated near doing, and adds acetonitrile and dissolves, and mixes, obtain need testing solution with 0.1% aqueous formic acid dilution.
10. high performance liquid chromatography-tandem mass method detects the method for sample Pesticide Residues, it is characterized in that: it comprises the following steps:
1., according to sample-pretreating method according to claim 9, need testing solution is obtained;
2., high performance liquid chromatography-tandem mass method is adopted to detect need testing solution:
High-efficient liquid phase chromatogram condition:
Chromatographic column: AgilentZORBAXRX-C8 post;
Mobile phase A: 0.1% formic acid-aqueous solution, Mobile phase B: acetonitrile;
Gradient elution program is as follows:
Tandem mass spectrum condition: electric spray ion source.
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