CN105032381B - Compound molecule trace solid-phase extraction column and preparation method and application - Google Patents
Compound molecule trace solid-phase extraction column and preparation method and application Download PDFInfo
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Abstract
The invention discloses the preparation method of compound molecule trace solid-phase extraction column, and it comprises the following steps：A, molecularly imprinted polymer A is prepared by template molecule of triazine；B, molecularly imprinted polymer B is prepared by template molecule of sulfonylurea pesticide；C, compound molecule trace solid-phase extraction column is prepared.The compound molecule trace solid-phase extraction column of the present invention, be capable of simultaneous selection enrichment, extraction sample in triazine and sulfonylurea pesticide, it is easy to use, enrichment, the effect of extraction are good, and can at least it reuse 20 times, with economic and environment-friendly, low cost and other advantages, it is especially suitable for simultaneously being enriched with triazine and sulfonylurea pesticide, extracts, when the compound molecule trace solid-phase extraction column of the present invention is used for the detection of high performance liquid chromatography tandem mass spectrum method, testing result is accurately and reliably.
Description
Technical field
The present invention relates to compound molecule trace solid-phase extraction column and preparation method and application.
Background technology
Triazines and sulfonylurea herbicide are a kind of herbicides for having hexa-member heterocycle structure, since release, have just been made
Worldwide widely used for highy potent herbicide to prevent farmland weed growth, in particular for cereal crops.Triazines
Herbicide is widely used all over the world, but residual of such herbicide in crops can bring certain harm to human body,
Its safety issue remained in crops has caused increasing concern.
Molecularly imprinted polymer (MIPs) can be in the absorption complex matrices of selectivity target compound, while have one
The stability of fixed mechanical strength and acid and alkali-resistance heat, and service life is long, affinity is high, selectivity is good, is that a kind of performance is excellent
Good SPE material, molecularly imprinted solid phase extraction column can be fabricated to and used.Compared with traditional SPE, point
Sub- trace SPE cost is cheap, simple to operate, while can effectively remove sample substrate interference, improves the accurate of analysis
Property, it is more suitable for trace analysis.
At present, disclosed molecularly imprinted solid phase extraction column is mainly unimolecule trace solid-phase extraction column, such as：Chinese CN
104001486 A hydrophily Sulfa drugs molecule trace solid-phase extraction column, the A of Chinese patent CN 102974327 amino
Glycoside drug molecule trace solid-phase extraction column, the A of Chinese patent CN 102631902 Coumarins rat poison molecular engram are consolidated
Phase extraction column, the unimolecule trace solid-phase extraction column can only be extracted to certain a kind of medicine, then tested and analyzed, without
It can realize that the medicine different to two classes is extracted, detected simultaneously；In the prior art, it there are no and utilize compound molecule trace solid phase
The relevant report that extraction column is extracted, detected to medicines more than two classes or two classes simultaneously.
Therefore, it is necessary to invent a kind of new molecularly imprinted solid phase extraction column, its can simultaneously to two classes or two classes more than
Medicine extracted, detected.
The content of the invention
It is an object of the invention to provide a kind of preparation method of compound molecule trace solid-phase extraction column.
A kind of preparation method of compound molecule trace solid-phase extraction column provided by the invention, it comprises the following steps：
A, molecularly imprinted polymer A is prepared by template molecule of triazine：
Triazine dissolves in organic solvent, adds function monomer A, concussion, adds crosslinking agent and initiator, surpasses
Sound, fill N2Deoxygenation, reacted after sealing, obtain polymer A；Polymer A is crushed, 200 mesh sieves is crossed, washes away template molecule,
Obtain molecularly imprinted polymer A；
Triazine, function monomer A, crosslinking agent, the mol ratio of initiator are 1：3~5：3~5：0.5~0.8；Triazine
The molal volume of class agricultural chemicals and organic solvent ratio is 1：8~12mmol/mL；
B, molecularly imprinted polymer B is prepared by template molecule of sulfonylurea pesticide：
Sulfonylurea pesticide dissolves in organic solvent, adds function monomer B, concussion, adds crosslinking agent and initiator, surpasses
Sound, fill N2Deoxygenation, reacted after sealing, obtain polymer B；Template molecule is washed away, obtains molecularly imprinted polymer B；
Sulfonylurea pesticide, function monomer B, crosslinking agent, the mol ratio of initiator are 1：3~5：3~5：0.5~0.8；Sulphur
The molal volume of ureide derivative agricultural chemicals and organic solvent ratio is 1：150~170mmol/mL；
C, compound molecule trace solid-phase extraction column is prepared：
Using molecularly imprinted polymer A and molecularly imprinted polymer B as filler, it is filled into solid phase of the bottom equipped with sieve plate and extracts
Take in post void column, upper end adds sieve plate, compresses, produces compound molecule trace solid-phase extraction column；Molecularly imprinted polymer A and molecule
Imprinted polymer B weight ratio is 1：4~6.
Further,
In step a, triazine is selected from ametryn, prometryn, cyanazine, cyprazine, Simanex, dipropetryn, methoxy
Third is net, symetryne, Gesadrual, atraton, Garagard, propazine, de- isopropyl-Atrazine, de- ethyl-Atrazine, spy
Any one or more in Ding Tong, prometon, Atrazine；Function monomer A is methacrylic acid；Crosslinking agent is trimethyl third
Alkane trimethyl acrylic ester；Initiator is azodiisobutyronitrile；Organic solvent is selected from acetonitrile or dichloromethane；
In step b, sulfonylurea pesticide is selected from chlorine sulphur is grand, any one or more in monosulfmeturon, thifensulfuronmethyl；Work(
Energy monomer B is diethylaminoethyl methacrylate；Crosslinking agent is trimethyl propane trimethyl acrylic ester；Initiator is azo
Bis-isobutyronitrile；Organic solvent is selected from acetonitrile or dichloromethane；
In step c, molecularly imprinted polymer A be selected from ametryn molecularly imprinted polymer, prometryn molecularly imprinted polymer,
Cyanazine molecularly imprinted polymer, cyprazine molecularly imprinted polymer, Simanex molecularly imprinted polymer, dipropetryn molecular engram
Polymer, the third net molecularly imprinted polymer of methoxy, symetryne molecularly imprinted polymer, Gesadrual molecularly imprinted polymer, green bristlegrass are gone
Logical 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 polymers, prometon molecule
Any one or more in imprinted polymer, Atrazine molecularly imprinted polymer；It is grand that molecularly imprinted polymer B is selected from chlorine sulphur
In molecularly imprinted polymer, monosulfmeturon molecularly imprinted polymer, thifensulfuronmethyl molecularly imprinted polymer any one or it is more
Kind.
Further, molecularly imprinted polymer A includes atrazine molecularly imprinted polymer and prometryn molecular engram polymerize
Thing；Molecularly imprinted polymer B is the grand molecularly imprinted polymer of chlorine sulphur.
Further,
In step a, triazine, function monomer A, crosslinking agent, the mol ratio of initiator are 1:4:4:0.6；Triazines
The molal volume of agricultural chemicals and organic solvent ratio is 1:10mmol/mL；
In step b, sulfonylurea pesticide, function monomer B, crosslinking agent, the mol ratio of initiator are 1:4:4:0.6；Sulphonyl
The molal volume of carbamide pesticide and organic solvent ratio is 1：160mmol/mL；
In step c, molecularly imprinted polymer A and molecularly imprinted polymer B weight ratio are 1:5.
Further, in molecularly imprinted polymer A, atrazine molecularly imprinted polymer and prometryn molecularly imprinted polymer
Weight ratio be 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；With
10% acetic acid methanol solution washes away template molecule in Soxhlet extraction device；
In step b, the concussion time is 30min；Reaction temperature is 50 DEG C~70 DEG C；Reaction time is 20h~28h；With
10% acetic acid methanol solution washes away template molecule in Soxhlet extraction device.
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 the above method.
A kind of compound molecule trace solid-phase extraction column provided by the invention, it is that the solid phase being prepared by the above method extracts
Take post.
Present invention also offers the sample-pretreating method of Detecting Pesticide, and it comprises the following steps：
I, the sample for taking homogeneous to prepare, mix, mechanical shaking extraction, centrifuge with acetonitrile or the acetonitrile solution of n-hexane saturation, weight
Multiple extraction twice, merges supernatant, as acetonitrile extract solution；
Ii, acetonitrile extract solution is taken, be concentrated to dryness, redissolved with acetonitrile, obtain sample liquid；
Iii, sampling liquid cross the compound molecule trace solid-phase extraction column of claim 8, coutroi velocity 0.3mL/min, stream
It is dry；With water wash, drain off；Methanol, acetate-methanol is used to elute successively, collection elution liquid nitrogen, which blows to be concentrated into, closely to be done, and adds acetonitrile molten
Solution, diluted and mixed with 0.1% aqueous formic acid, obtain need testing solution.
Further,
In step i, the sample is plant-derived food；The w/v of sample and acetonitrile is 2:15g/ml；Vibration
The time of extraction is 10min；The rotating speed of centrifugation is 4000r/min, and the time of centrifugation is 5min；
In step ii, it is concentrated to dryness using parallel quantitative concentrating instrument；Temperature during concentration is 50 DEG C；
In step iii, sample liquid, leaching scouring water, methanol, acetate-methanol, acetonitrile, the volume ratio of 0.1% aqueous formic acid are
1:1:2:6:1:1；In acetate-methanol, the volume ratio of acetic acid and methanol is 1：9.
Present invention also offers the method for high performance liquid chromatography-tandem mass method detection sample Pesticide Residues, it includes
Following steps：
1., according to the sample-pretreating method described in claim 9, obtain need testing solution；
2., using high performance liquid chromatography-tandem mass method need testing solution is detected：
High-efficient liquid phase chromatogram condition：
Chromatographic column：Agilent ZORBAX RX-C8 posts；
Mobile phase A：0.1% formic acid-the 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, and flow velocity is 200 μ L/min, and sample size is 3 μ L；Series connection
Mass Spectrometry Conditions：Ion source temperature is 450 DEG C, and cation scanning, multiple-reaction monitoring pattern detection, atomization gas pressure is 30psi, is added
Hot assist gas pressure power is 30psi, and gas curtain atmospheric pressure is 35psi, spray voltage 5500V.
The present invention compound molecule trace solid-phase extraction column, be capable of simultaneous selection enrichment, extraction sample in triazine
Class agricultural chemicals and sulfonylurea pesticide, easy to use, enrichment, the effect of extraction are good, and can at least reuse 20 times, have warp
Ji environmental protection, low cost and other advantages, are especially suitable for simultaneously being enriched with triazine and sulfonylurea pesticide, extract, this hair
When bright compound molecule trace solid-phase extraction column is used for the detection of high performance liquid chromatography-tandem mass method, testing result is accurately and reliably.
Obviously, according to the above of the present invention, according to the ordinary technical knowledge and customary means of this area, do not departing from
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The embodiment of form by the following examples, the above of the present invention is remake further specifically
It is bright.But the scope that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to following example.It is all to be based on the above of the present invention
The technology realized belongs to the scope of the present invention.
Brief description of the drawings
Fig. 1 is that the SEM of atrazine molecularly imprinted polymer of the present invention schemes；
Fig. 2 is that the SEM of prometryn molecularly imprinted polymer of the present invention schemes；
Fig. 3 is that the SEM of the grand molecularly imprinted polymer of chlorine sulphur of the present invention schemes；
Fig. 4 be the μ g/kg of mark-on level 10 corn sample in 20 kinds of agricultural chemicals total ion current (total ion current,
TIC) chromatogram；
Fig. 5 is the grand mass spectrogram of chlorine sulphur.
Fig. 6 is the mass spectrogram of de- isopropyl-Atrazine；
Fig. 7 is the total ion chromatogram of typical positive sample.
Embodiment
The raw material that is used in the specific embodiment of the invention, equipment are known product, are obtained by buying commercially available prod.
Embodiment 1
1.1st, instrument and reagent
AB5500 high performance liquid chromatography-triple quadrupole rods tandem mass spectrometries instrument, equipped with electric spray ion source (ESI) and
Analyst1.6.2 work stations (American AB SCIEX companies)；Parallel quantitative concentrating instrument, B-740 recirculated cooling systems, V-
700/701 vavuum pump (BUCHI companies of Switzerland)；Nitrogen evaporator (Biotage companies of Sweden)；(U.S. Beckman is public for supercentrifuge
Department)；
Acetonitrile, methanol (chromatographically pure, Fisher companies), n-hexane (chromatographically pure, Honeywell companies), formic acid (chromatogram
It is pure, German CNW companies)；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), ring
Third Tianjin (cyprazine), Simanex (simazine), dipropetryn (dipropetryn), methoxy third net (methoprotryne),
Symetryne (simetryn), Gesadrual (simeton), atraton (atraton), Garagard (terbuthylazine), put out
Tianjin (propazine), de- isopropyl-Atrazine (atrazine-desisopropyl), de- ethyl-Atrazine
(atrazine-desethyl), Te Dingtong (terbumeton), prometon (prometon), Atrazine (atrazine), chlorine
Sulphur grand (chlorsulfuron), monosulfmeturon (monosulfuron), thifensulfuronmethyl (thifensulfuron-methyl) are (pure
Degree is all higher than 97%, is purchased from German Dr.Ehrenstorfer companies)；
Experimental water is Milli-Q water (distilled water through Millipore, Bedford, MA, after USA production water purification machine processing
Ultra-pure water).
1.2nd, the preparation of standard liquid
Single standard storing solution：The standard sample of pesticide of 5mg (being accurate to 0.01mg) is accurately weighed respectively, is placed in 10mL capacity
In bottle, dissolved with methanol and dilute constant volume, be made into the standard reserving solution that mass concentration is 500 μ g/mL, lucifuge is stored up at -18 DEG C
Deposit.
Mixed standard solution：Accurate each pesticide standard stock solutions of 1mL of drawing are determined into 50mL volumetric flasks with dilution in acetonitrile
Hold, be configured to the hybrid standard working solution that each pesticide composition is 10 μ g/mL, be kept in dark place under the conditions of 4 DEG C.First during use
Alcohol is diluted to finite concentration, carries out high performance liquid chromatography-tandem mass method detection.
1.3rd, the preparation of the SPE of compound molecule trace (CMISPE) post
A, the preparation of triazine molecularly imprinted polymer：
Prometryn (0.2mmol) or atrazine (0.2mmol) are dissolved in the acetonitrile or dichloromethane (2mL) of certain volume
In, function monomer MAA (methacrylic acid, 0.8mmol) is added thereto, is shaken 30min, is then added a certain amount of crosslinking agent
TRIM (trimethyl propane trimethyl acrylic ester, 0.8mmol) and initiator A IBN (azodiisobutyronitrile, 20mg), ultrasound
5min, fill N2Deoxygenation 10min, it is put into after sealing in water-bath and 24h is reacted at 60 DEG C, is taken out polymer after the completion of reaction, grind
200 mesh sieves are crossed after mill, template is washed away in Soxhlet extraction device with 10% acetic acid methanol solution, atrazine molecular engram is obtained and gathers
Compound, prometryn molecularly imprinted polymer；Its ESEM testing result is shown in Fig. 1 and Fig. 2 respectively.
B, the preparation of sulfonylurea pesticide molecularly imprinted polymer：
Chlorine sulphur grand (0.2mmol) is dissolved in the acetonitrile (32mL) of certain volume, adds function monomer DEMEA (methyl-props
Olefin(e) acid lignocaine ethyl ester, 0.8mmol), 30min is shaken, a certain amount of crosslinking agent TRIM (0.8mmol) is then added and triggers
Agent AIBN 20mg, ultrasound, fill N2Deoxygenation, it is put into after sealing in water-bath and 24h is reacted at 60 DEG C, is taken out after the completion of reaction poly-
Compound, template is washed away in Soxhlet extraction device with 10% acetic acid methanol, obtain the grand molecularly imprinted polymer of chlorine sulphur；It scans electricity
Mirror testing result is shown in Fig. 3.
C, the preparation of compound molecule trace solid-phase extraction column (CMISPE posts)：
Weigh 50mg atrazines molecularly imprinted polymer, 30mg prometryns molecularly imprinted polymer and grand point of 400mg chlorine sulphurs
Sub- imprinted polymer mix suspending is filled into solid-phase extraction column void column of the bottom equipped with sieve plate, upper end adds in acetonitrile
CMISPE posts are made in sieve plate, compression.
1.4th, the sample-pretreating method of Detecting Pesticide
Extraction：
The accurate sample 2.0g for weighing homogeneous preparation is placed in 50mL centrifuge tubes, is added 15mL acetonitriles, is vortexed and mixes 1min,
Mechanical shaking extraction 10min again, 4000r/min centrifuge 5min, repeat extraction twice, merge supernatant (for fat contents such as corns
Higher sample, (it is n-hexane to remove layer solution after n-hexane, acetonitrile mixing to the acetonitrile solution of addition 20mL n-hexane saturations
The acetonitrile solution of saturation) mechanical shaking extraction, discard n-hexane layer), obtain acetonitrile extract solution.The parallel quantitative concentration of acetonitrile extract solution
Instrument (temperature adjustment is to 50 DEG C) is concentrated to dryness, and is redissolved with 2mL acetonitriles, is taken wherein 1mL sample liquids, to be clean.
SPE：
CMISPE posts are activated with 3mL acetonitriles, acetonitrile is drained off；1mL sample liquids are added in solid-phase extraction column, controlling stream
Speed about 0.3mL/min, drains off；With 1mL water wash, drain off；Successively with 2 × 1mL methanol and 6 × 1mL acid alcohol solution (acetic acid：First
Alcohol (V/V)=1：9) elute, collection elution liquid nitrogen, which blows to be concentrated into, closely to be done, and adds 1.0mL acetonitriles to dissolve, with the formic acid of mobile phase 0.1%
The aqueous solution 1:1 dilution is mixed to 2.0mL, as need testing solution, upper machine analysis.
Regeneration：
With 4mL acid alcohol solution (acetic acid：Methanol (V/V)=1：9) CMISPE posts are eluted more than 3 times, is washed out to without object
Afterwards, pillar may be reused more than 20 times.
1.5th, need testing solution is detected using high performance liquid chromatography-tandem mass method：
High-efficient liquid phase chromatogram condition：
Chromatographic column：(i.d represents the interior of chromatographic column to Agilent ZORBAX RX-C8 posts (150mm × 2.1mm i.d, 5 μm)
Footpath)；Mobile phase A：0.1% formic acid-the aqueous solution, Mobile phase B：Acetonitrile；Gradient elution program：It is shown in Table 1；Flow velocity：200 μ L/min,；
Column temperature：30℃；Sample size：3μL.
The gradient elution program of table 1, 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℃；Cation scans；Multiple-reaction monitoring (MRM) pattern is examined
Survey；Atomization gas (Gas1) pressure：30psi；Heating auxiliary gas (Gas2) pressure：30psi；Gas curtain gas (CUR) pressure：35psi；Spray
Mist voltage (IS):5500V；Remaining mass spectrometry parameters is shown in Table 2 and (20ng/mL mixed standard solution is taken, using the side of pin pump sample introduction
Formula, determine the mass spectrometry parameters of each compound).
The Mass Spectrometer Method condition and retention time of table 2,20 kind of herbicide
Note：* it is quota ion
The compound molecule trace solid-phase extraction column of the present invention, has advantages below：
(1), the present invention is first by triazine herbicide molecularly imprinted polymer and sulfonylurea weedicide molecularly imprinted poly-
Compound mixing is filled into solid-phase extraction column, is enriched with while realizing 17 kinds of triazines and 3 kinds of sulfonylurea pesticides；
(2), compound molecule trace solid-phase extraction column of the invention, two kinds of molecularly imprinted polymers are mixed as filler,
Can solve the problems, such as that current universal solid-phase extraction column poor selectivity, matrix effect are big, can also solve current molecular trace
The problem of solid-phase extraction column bound species are few, trace triazine and sulphur that can be in selective cleaning, enriched sample
Ureide derivative agricultural chemicals, to two class herbicides totally 20 kinds of materials the rate of recovery be 62.7%~117.4%, meet trace residue detection
It is required that；
(3), compared with universal solid-phase extraction column, compound molecule trace solid phase column of the invention, pretreatment process is simple,
Step is few, the effect of enrichment purification is good, and organic reagent usage amount is few, economic and environment-friendly, and at least may be reused after regenerating
20 times.
In order to illustrate beneficial effects of the present invention, the present invention provides tests below example：
The screening of the Extraction solvent of test example 1
Compare 4 kinds of acetonitrile, methanol, acetone, ethyl acetate solvents and the extraction that concentration is 10 μ g/kg is added in corn sample
Efficiency.
As a result find, using methanol as Extraction solvent, its corresponding rate of recovery is 43%~78%；Extraction is used as using acetone
It is 45%~88% during solvent；As 58.4%~109.8% during using ethyl acetate as Extraction solvent；Moreover, acetone, methanol carry
Take pigment content in liquid higher, the boiling point of ethyl acetate is higher, and concentration is time-consuming too long, is unsuitable for being used as Extraction solvent.
When the present invention uses acetonitrile as Extraction solvent, extraction efficiency is 79%~116%, and extraction effect is also more managed
Think.
The screening of the purification method of test example 2
Purification method includes liquid-liquid extraction and molecular engram solid phase extraction.The sample fat content such as corn and soybean is more, meeting
Cause extract solution muddy, n-hexane is usually used in extracting degreasing, therefore is directed to such sample, selects n-hexane to carry out liquid after extracted
Liquid extracts, and can remove most grease class chaff interference, then thorough to purify after post.Have in view of n-hexane in acetonitrile
Certain solubility, to ensure the rate of recovery, thus purified from the n-hexane of acetonitrile saturation.
The screening test of the SPE of test example 3
Acetonitrile, methanol, acid alcohol (acetic acid are investigated：Methanol (V/V)=1：9) with 4 kinds of different adsorption solvents of water to adsorptivity
Can influence, use above-mentioned 4 kinds different eluent solvent compound molecule imprinting solid phase extraction small columns after loading, in detection leacheate
The rate of recovery situation of 20 kinds of pesticides.
As a result show, it is preferable to most of herbicide absorption property when water and acetonitrile are as adsorbing medium, in leacheate
Almost without target detection thing, but the partial loss of monosulfmeturon and thifensulfuronmethyl can be caused in Acetonitrile, thus this research
Using water as eluent.Because the hydrogen bond action constant of methanol is much larger than acetonitrile, so imprinted polymer is in METHANOL MEDIUM
Absorption property it is minimum, thus have selected methanol as eluting solvent, Hyarogen-bonding can further be destroyed by adding acetic acid, be made
Target compound elution is complete.Experiment compares methanol, acid alcohol and both eluting powers when being used in combination, as a result find according to
It is secondary to reach optimal elution effect using methanol, acid alcohol, and investigated influence of the elution volume to the rate of recovery, as a result table
Bright, as use methanol 2mL, acid alcohol 6mL, target compound elution is complete, it is ensured that enough rate of recovery.
The screening of the high performance liquid chromatography-tandem mass method testing conditions of test example 4
Sulfonylurea herbicide is in faintly acid, and mobile phase is acidified through acid or glacial acetic acid, and adds 0.1% formic acid more in water
Be advantageous to the ionization of compound, compound peak shape can be improved.Select herein using the formic acid of acetonitrile -0.1% as mobile phase,
The separation detection of 20 kinds of agricultural chemicals is completed in 13min, and peak shape is sharp, separating degree is preferable.
The total ion current (total ion current, TIC) of 20 kinds of agricultural chemicals in the μ g/kg of mark-on level 10 corn sample
Chromatogram, see Fig. 4.
The grand mass spectrogram of chlorine sulphur, 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
It is determined that under optimum analysis condition, the mixed standard solution of various concentrations is subjected to HPLC-MS/MS analyses.Respectively
Using testing concentration as abscissa, peak area is that ordinate carries out linear regression.
As a result show：20 kinds of herbicides linear relationship in 0.5~20ng/mL concentration ranges is good, and coefficient correlation is
0.9942~0.9999, the requirement of quantitative analysis can be met.The detection for determining method with S/N >=3 is limited to 0.057~0.63 μ
G/kg, determine that quantifying for method is limited to 0.19~2.1 μ g/kg with S/N >=10.The line of 20 kinds of triazines and sulfonylurea herbicide
Sexual intercourse, the range of linearity and quantitative limit are shown 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 10 4 powers, and other are similar.
The veracity and precision of the method for test example 6
Mixed standard solution is added in blank corn sample makes final concentration of 5 μ g/kg, 10 μ g/kg, 20 μ g/kg, according to
1.4 carry out the pre-treatment of samples, each concentration level replication 6 times；Average recovery rate and precision the results are shown in Table 2.
As a result show, the average recovery rate of each compound is between 62.7%~117.4%, relative standard deviation (RSD)
For 1.7%~13.9%, meet the technical requirements of food Physico-chemical tests method confirmation.
According to national standard GB 2763-2014《Pesticide MRL》And Hong Kong《The remaining rule of pesticide in food
Example》Select 10 kinds of different samples, it detected using this method, the results are shown in Table 4, respectively corn, glutinous corn,
6 kinds of residues of pesticides are detected altogether in 5 kinds of brown rice, pineapple, lemon samples.
The chromatogram of typical positive sample, is shown in Fig. 7.
Table 4, the testing result for not planting 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 | ND | ND | ND | ND | ND | ND |
ND：Do not detect (not detected).
In summary, compound molecule trace solid-phase extraction column of the invention, it is capable of enrichment, the extraction sample of simultaneous selection
In triazine and sulfonylurea pesticide, easy to use, enrichment, the effect of extraction are good, and can at least reuse 20
It is secondary, there are economic and environment-friendly, low cost and other advantages, be especially suitable for simultaneously being enriched with triazine and sulfonylurea pesticide,
Extraction, when compound molecule trace solid-phase extraction column of the invention is used for the detection of high performance liquid chromatography-tandem mass method, testing result
Accurately and reliably.
Claims (7)
- A kind of 1. preparation method of compound molecule trace solid-phase extraction column, it is characterised in that：It comprises the following steps：A, molecularly imprinted polymer A is prepared by template molecule of triazine：Triazine dissolves in organic solvent, adds function monomer A, concussion, adds crosslinking agent and initiator, ultrasound, fill N2 Deoxygenation, reacted after sealing, obtain polymer A；Polymer A is crushed, 200 mesh sieves is crossed, washes away template molecule, obtain molecule Imprinted polymer A；Triazine, function monomer A, crosslinking agent, the mol ratio of initiator are 1:4:4:0.6；Triazine with it is organic molten The molal volume ratio of agent is 1:10 mmol/mL；B, molecularly imprinted polymer B is prepared by template molecule of sulfonylurea pesticide：Sulfonylurea pesticide dissolves in organic solvent, adds function monomer B, concussion, adds crosslinking agent and initiator, ultrasonic, Fill N2Deoxygenation, reacted after sealing, obtain polymer B；Template molecule is washed away, obtains molecularly imprinted polymer B；Sulfonylurea pesticide, function monomer B, crosslinking agent, the mol ratio of initiator are 1:4:4:0.6；Sulfonylurea pesticide is with having The molal volume ratio of solvent is 1：160 mmol/mL；C, compound molecule trace solid-phase extraction column is prepared：Using molecularly imprinted polymer A and molecularly imprinted polymer B as filler, the solid-phase extraction column that bottom is equipped with sieve plate is filled into In void column, upper end adds sieve plate, compresses, produces compound molecule trace solid-phase extraction column；Molecularly imprinted polymer A and molecular engram The weight ratio of polymer B is 1:5；In step a, triazine be selected from ametryn, prometryn, cyanazine, cyprazine, Simanex, dipropetryn, methoxy third it is net, Symetryne, Gesadrual, atraton, Garagard, propazine, de- isopropyl-Atrazine, de- ethyl-Atrazine, Te Dingtong, Any one or more in prometon, Atrazine；Function monomer A is methacrylic acid；Crosslinking agent is trimethyl propane three Methacrylate；Initiator is azodiisobutyronitrile；Organic solvent is selected from acetonitrile or dichloromethane；In step b, sulfonylurea pesticide is selected from chlorine sulphur is grand, any one or more in monosulfmeturon, thifensulfuronmethyl；Function list Body B is diethylaminoethyl methacrylate；Crosslinking agent is trimethyl propane trimethyl acrylic ester；Initiator is that azo two is different Butyronitrile；Organic solvent is selected from acetonitrile or dichloromethane；In step c, molecularly imprinted polymer A is selected from ametryn molecularly imprinted polymer, prometryn molecularly imprinted polymer, cyanogen grass Tianjin molecularly imprinted polymer, cyprazine molecularly imprinted polymer, Simanex molecularly imprinted polymer, the polymerization of dipropetryn molecular engram Thing, the third net molecularly imprinted polymer of methoxy, symetryne molecularly imprinted polymer, Gesadrual molecularly imprinted polymer, atraton point Sub- imprinted polymer, Garagard molecularly imprinted polymer, propazine molecularly imprinted polymer, de- isopropyl-Atrazine molecule Imprinted polymer, de- ethyl-Atrazine molecularly imprinted polymer, Te Dingtong molecularly imprinted polymers, prometon molecular engram Any one or more in polymer, Atrazine molecularly imprinted polymer；Molecularly imprinted polymer B is selected from the grand molecule of chlorine sulphur Any one or more in imprinted polymer, monosulfmeturon molecularly imprinted polymer, thifensulfuronmethyl molecularly imprinted polymer.
- 2. preparation method according to claim 1, it is characterised in that：Molecularly imprinted polymer A prints including atrazine molecule Mark polymer and prometryn molecularly imprinted polymer；Molecularly imprinted polymer B is the grand molecularly imprinted polymer of chlorine sulphur.
- 3. preparation method according to claim 1, it is characterised in that：In molecularly imprinted polymer A, atrazine molecular engram The weight of polymer and prometryn molecularly imprinted polymer ratio is 5:3.
- 4. according to the preparation method described in claims 1 to 3 any one, it is characterised in that：In step a, the concussion time is 30min；Reaction temperature is 50 DEG C~70 DEG C；Reaction time is 20h~28h；With for 10% second Sour methanol solution washes away template molecule in Soxhlet extraction device；In step b, the concussion time is 30min；Reaction temperature is 50 DEG C~70 DEG C；Reaction time is 20h~28h；With for 10% second Sour methanol solution washes away template molecule in Soxhlet extraction device.
- 5. preparation method according to claim 4, it is characterised 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.
- A kind of 6. compound molecule trace solid-phase extraction column, it is characterised in that：It is by the method for Claims 1 to 5 any one The solid-phase extraction column being prepared.
- 7. the sample-pretreating method of Detecting Pesticide, it is characterised in that：It comprises the following steps：I, the sample for taking homogeneous to prepare, mixed with acetonitrile or the acetonitrile solution of n-hexane saturation, mechanical shaking extraction, centrifugation, repetition carries Take twice, merge supernatant, as acetonitrile extract solution；Ii, acetonitrile extract solution is taken, be concentrated to dryness, redissolved with acetonitrile, obtain sample liquid；Iii, sampling liquid cross the compound molecule trace solid-phase extraction column of claim 6, coutroi velocity 0.3mL/min, drain off；With Water wash, drain off；Eluted successively with methanol, acetate-methanol, collect elution liquid nitrogen and blow and be concentrated near dry, add acetonitrile to dissolve, with for The dilution of 0.1% aqueous formic acid mixes, and obtains need testing solution.
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