CN114062576B - Molecularly imprinted electrostatic spinning fiber membrane, preparation method thereof, microfluidic chip and method for detecting zearalenone - Google Patents
Molecularly imprinted electrostatic spinning fiber membrane, preparation method thereof, microfluidic chip and method for detecting zearalenone Download PDFInfo
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- CN114062576B CN114062576B CN202111529848.XA CN202111529848A CN114062576B CN 114062576 B CN114062576 B CN 114062576B CN 202111529848 A CN202111529848 A CN 202111529848A CN 114062576 B CN114062576 B CN 114062576B
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- fiber membrane
- zearalenone
- electrostatic spinning
- molecularly imprinted
- pillow
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- 239000000835 fiber Substances 0.000 title claims abstract description 116
- 239000012528 membrane Substances 0.000 title claims abstract description 107
- 238000010041 electrostatic spinning Methods 0.000 title claims abstract description 84
- MBMQEIFVQACCCH-QBODLPLBSA-N zearalenone Chemical compound O=C1O[C@@H](C)CCCC(=O)CCC\C=C\C2=CC(O)=CC(O)=C21 MBMQEIFVQACCCH-QBODLPLBSA-N 0.000 title claims abstract description 77
- MBMQEIFVQACCCH-UHFFFAOYSA-N trans-Zearalenon Natural products O=C1OC(C)CCCC(=O)CCCC=CC2=CC(O)=CC(O)=C21 MBMQEIFVQACCCH-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 238000000034 method Methods 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 229920002239 polyacrylonitrile Polymers 0.000 claims abstract description 38
- 238000001179 sorption measurement Methods 0.000 claims abstract description 20
- 238000001514 detection method Methods 0.000 claims abstract description 11
- 238000006243 chemical reaction Methods 0.000 claims description 43
- 239000000243 solution Substances 0.000 claims description 35
- 229920000344 molecularly imprinted polymer Polymers 0.000 claims description 31
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 238000009987 spinning Methods 0.000 claims description 29
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 27
- 229920003229 poly(methyl methacrylate) Polymers 0.000 claims description 26
- 239000004926 polymethyl methacrylate Substances 0.000 claims description 26
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 22
- 239000000758 substrate Substances 0.000 claims description 20
- 238000002156 mixing Methods 0.000 claims description 19
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims description 15
- 238000002347 injection Methods 0.000 claims description 15
- 239000007924 injection Substances 0.000 claims description 15
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- 239000003480 eluent Substances 0.000 claims description 13
- 238000006116 polymerization reaction Methods 0.000 claims description 13
- KYITYFHKDODNCQ-UHFFFAOYSA-M sodium;2-oxo-3-(3-oxo-1-phenylbutyl)chromen-4-olate Chemical compound [Na+].[O-]C=1C2=CC=CC=C2OC(=O)C=1C(CC(=O)C)C1=CC=CC=C1 KYITYFHKDODNCQ-UHFFFAOYSA-M 0.000 claims description 13
- 229960002647 warfarin sodium Drugs 0.000 claims description 13
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 claims description 10
- OKKRPWIIYQTPQF-UHFFFAOYSA-N Trimethylolpropane trimethacrylate Chemical compound CC(=C)C(=O)OCC(CC)(COC(=O)C(C)=C)COC(=O)C(C)=C OKKRPWIIYQTPQF-UHFFFAOYSA-N 0.000 claims description 9
- OZAIFHULBGXAKX-UHFFFAOYSA-N 2-(2-cyanopropan-2-yldiazenyl)-2-methylpropanenitrile Chemical compound N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 claims description 8
- 239000007791 liquid phase Substances 0.000 claims description 6
- 238000004949 mass spectrometry Methods 0.000 claims description 5
- 239000012046 mixed solvent Substances 0.000 claims description 5
- 229930191978 Gibberellin Natural products 0.000 claims description 3
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 claims description 3
- 239000003448 gibberellin Substances 0.000 claims description 3
- 230000035484 reaction time Effects 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 9
- 230000035945 sensitivity Effects 0.000 abstract description 9
- 239000000463 material Substances 0.000 abstract description 7
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
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- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 4
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- 239000000178 monomer Substances 0.000 description 3
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- 239000000126 substance Substances 0.000 description 3
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 2
- 241000223218 Fusarium Species 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
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- 108700012359 toxins Proteins 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 229930195730 Aflatoxin Natural products 0.000 description 1
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 description 1
- 235000007319 Avena orientalis Nutrition 0.000 description 1
- 241000209763 Avena sativa Species 0.000 description 1
- 235000007558 Avena sp Nutrition 0.000 description 1
- 241000223195 Fusarium graminearum Species 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 231100000678 Mycotoxin Toxicity 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 244000062793 Sorghum vulgare Species 0.000 description 1
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
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- 230000001076 estrogenic effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- -1 fusarium graminearum Chemical compound 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
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- 150000003431 steroids Chemical class 0.000 description 1
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- 239000013076 target substance Substances 0.000 description 1
- 229960005080 warfarin Drugs 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/08—Preparation using an enricher
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/4282—Addition polymers
- D04H1/43—Acrylonitrile series
-
- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/4382—Stretched reticular film fibres; Composite fibres; Mixed fibres; Ultrafine fibres; Fibres for artificial leather
- D04H1/43838—Ultrafine fibres, e.g. microfibres
-
- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Textile Engineering (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Dispersion Chemistry (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Spinning Methods And Devices For Manufacturing Artificial Fibers (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention provides a molecular imprinting electrostatic spinning fiber membrane and a preparation method thereof, a microfluidic chip and a method for detecting zearalenone, and relates to the technical field of biotoxin analysis and detection. The molecular imprinting electrostatic spinning fiber membrane provided by the invention comprises a polyacrylonitrile nano electrostatic spinning fiber membrane and a zearalenone molecular imprinting polymer dispersed in the fibers of the polyacrylonitrile nano electrostatic spinning fiber membrane. The molecular imprinting electrostatic spinning fiber membrane provided by the invention has the characteristics of specific recognition of a molecular imprinting material, high specific surface area and sensitivity of an electrostatic nanofiber membrane, and not only can be used for specifically recognizing zearalenone, but also has the advantages of strong sensitivity, high adsorption efficiency, simple preparation, low cost and good stability. According to the invention, the molecular imprinting electrostatic spinning fiber membrane is combined with the microfluidic technology, and the obtained microfluidic chip can be used for simply, rapidly and sensitively detecting the zearalenone, and is low in cost.
Description
Technical Field
The invention relates to the technical field of biotoxin analysis and detection, in particular to a molecular imprinting electrostatic spinning fiber membrane and a preparation method thereof, a microfluidic chip and a method for detecting zearalenone.
Background
Zearalenone, also called F-2 toxin, is a non-steroid class with estrogenic activity mainly produced by fusarium, such as fusarium graminearum, fusarium trilineum and the like, mainly pollutes corn, wheat, rice, barley, millet, oat and other grains, can directly enter human bodies and animals through the crops such as the polluted grains and the like, can also enter human bodies through animal foods such as polluted meat, milk and the like, and is one of toxins which are seriously polluted in mycotoxins and have great harm to the human bodies and the animals. Therefore, monitoring control of zearalenone is very necessary.
At present, the sample is often required to be pretreated in the detection of zearalenone, a more common sample pretreatment method is an immunoaffinity column method, and the immunoaffinity column is a new technology applied to the analysis field in the 90 th century, and is prepared by coupling a certain amount of monoclonal antibodies on a proper carrier. The national standard GB 5009.209-2016 "determination of zearalenone in food safety national standard food" adopts an immunoaffinity column method, wherein zearalenone in a sample is extracted by acetonitrile solution, purified and enriched by the immunoaffinity column, and then eluted by water and methanol, and eluent is collected for liquid chromatography determination. The immunoaffinity column has the characteristic of high selective adsorption, the purification effect is good, the detection sensitivity is high, but the antibody of the immunoaffinity column is difficult to prepare, the period is long, the product price is high, and the detection cost is high.
Disclosure of Invention
In view of the above, the invention aims to provide a molecularly imprinted electrostatic spinning fiber membrane, a preparation method thereof, a microfluidic chip and a method for detecting zearalenone. The molecular imprinting electrostatic spinning fiber membrane provided by the invention not only can specifically identify zearalenone and has high sensitivity, but also has the advantages of simple preparation, low cost and good stability; the microfluidic chip combined with the chip technology has the characteristics of high mass transfer rate and low reagent consumption, and can be used for simply, rapidly and sensitively detecting the zearalenone with low cost.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a molecularly imprinted electrostatic spinning fiber membrane, which comprises a polyacrylonitrile nano electrostatic spinning fiber membrane and a zearalenone molecularly imprinted polymer dispersed in the fibers of the polyacrylonitrile nano electrostatic spinning fiber membrane.
Preferably, the mass content of the zearalenone molecularly imprinted polymer in the molecularly imprinted electrostatic spinning fiber membrane is 2-5%; the fiber diameter of the molecular imprinting electrostatic spinning fiber membrane is 279-311 nm.
The invention provides a preparation method of the molecular imprinting electrostatic spinning fiber membrane, which comprises the following steps:
mixing a zearalenone molecularly imprinted polymer and a polyacrylonitrile spinning solution to obtain a molecularly imprinted electrostatic spinning solution;
and carrying out electrostatic spinning on the molecular imprinting electrostatic spinning solution to obtain the molecular imprinting electrostatic spinning fiber membrane.
Preferably, the mass concentration of the polyacrylonitrile spinning solution is 8-12%; the dosage ratio of the zearalenone molecularly imprinted polymer to the polyacrylonitrile spinning solution is 20-25 mg: 1-2 mL.
Preferably, the preparation method of the zearalenone molecularly imprinted polymer comprises the following steps:
mixing warfarin sodium, acetonitrile and N-vinyl pyrrolidone for prepolymerization to obtain a prepolymerized product;
mixing the pre-polymerized product, trimethylolpropane trimethacrylate and azodiisobutyronitrile, and carrying out polymerization reaction at 60 ℃ to obtain a polymerized product;
eluting the polymerization product to obtain a zearalenone molecularly imprinted polymer;
the molar ratio of the warfarin sodium to the N-vinyl pyrrolidone to the trimethylolpropane trimethacrylate is 1:4:10; the eluting reagent is a mixed solvent of methanol and acetic acid.
Preferably, the spinning distance of the electrostatic spinning is 10-15 cm, the spinning voltage is 13-17 kV, and the flow rate is 20-50 mu L/min.
The invention provides a microfluidic chip, which comprises a PMMA chip substrate 1, a microfluidic channel 2 arranged in the PMMA chip substrate 1 and a plurality of pillow-shaped reaction tanks 3 arranged on the microfluidic channel 2; the microfluidic channel 2 is divided into a plurality of sections by the pillow-shaped reaction tank;
each pillow-shaped reaction tank 3 is filled with a fiber membrane 4; the fiber membrane 4 is the molecular imprinting electrostatic spinning fiber membrane described in the technical scheme or the molecular imprinting electrostatic spinning fiber membrane prepared by the preparation method described in the technical scheme;
sample injection holes 2-1 and sample discharge holes 2-2 are respectively arranged at two ports of the microfluidic channel 2, and the sample injection holes 2-1 and the sample discharge holes 2-2 penetrate through the upper surface of the PMMA chip substrate 1.
Preferably, the length of the microfluidic chip is 5cm, and the width is 3cm; the width of the micro-flow channel 2 is 1mm, and the depth is 2mm; the length of a single pillow-shaped reaction tank is 10mm, the width is 5mm, the depth is 2mm, and the number of the pillow-shaped reaction tanks 3 is 4; the length of the fiber membrane 4 is 10mm, and the width is 5mm; the radiuses of the sample inlet hole 2-1 and the sample outlet hole 2-2 are both 0.6mm.
A method of pre-treating a zearalenone-containing sample comprising the steps of:
leading sample liquid to be detected into a microfluidic chip through a sample inlet hole 2-1, flowing through each pillow-shaped reaction tank 3 through a microfluidic channel 2, adsorbing gibberellin in the sample liquid step by step through a fiber membrane 4 in each pillow-shaped reaction tank 3, and flowing out of the sample outlet hole 2-2;
after the adsorption is finished, introducing an eluting reagent into the microfluidic chip through the sample inlet holes 2-1, flowing through each pillow-shaped reaction tank 3 through the microfluidic channel 2, gradually desorbing the fiber membrane 4 in each pillow-shaped reaction tank 3, flowing out eluent through the sample outlet holes 2-2, and collecting the eluent; the eluting reagent is methanol.
A method for detecting zearalenone comprises the step of carrying out liquid phase mass spectrometry detection on the eluent in the technical scheme to obtain quantitative data of zearalenone.
The invention provides a molecularly imprinted electrostatic spinning fiber membrane, which comprises a polyacrylonitrile nano electrostatic spinning fiber membrane and a zearalenone molecularly imprinted polymer dispersed in the fibers of the polyacrylonitrile nano electrostatic spinning fiber membrane. The molecularly imprinted polymer in the molecularly imprinted electrostatic spinning fiber membrane provided by the invention can specifically identify the target substance zearalenone, is not limited by external conditions, and has the characteristics of specificity, good stability and low cost; the nano electrostatic spinning fiber membrane has the characteristics of large specific surface area and multiple pores, and can enable more molecular imprinting binding sites to be exposed by combining with the molecular imprinting polymer, so that the efficiency of grabbing and enriching target objects is improved. The molecular imprinting electrostatic spinning fiber membrane provided by the invention has the characteristics of specific recognition of a molecular imprinting material, high specific surface area and sensitivity of an electrostatic nanofiber membrane, and not only can the zearalenone be specifically recognized, but also the cost is low and the stability is good. The example result shows that the adsorption rate of the molecular imprinting electrostatic spinning fiber membrane provided by the invention to zearalenone is 75.56-94.50%.
The invention provides the preparation method of the molecular imprinting electrostatic spinning fiber membrane, which has the advantages of simple process, easy operation and contribution to realizing large-scale production.
The invention provides a microfluidic chip, which comprises a PMMA chip substrate, a microfluidic channel arranged in the PMMA chip substrate and a plurality of pillow-shaped reaction tanks arranged on the microfluidic channel, wherein the PMMA chip substrate is provided with a plurality of grooves; each pillow-shaped reaction tank is filled with a fiber membrane, and the fiber membrane is the molecular imprinting electrostatic spinning fiber membrane according to the technical scheme. The chip is used as a carrier of the molecular imprinting electrostatic spinning fiber membrane, the rigidity of the chip is high, and the problems that the molecular imprinting electrostatic spinning fiber membrane is high in electrostatic adsorption force, fragile in membrane body and difficult to fix can be solved; moreover, the microfluidic channel has narrow space, high mass transfer rate and less consumption of reagents; the molecular imprinting polymer is used as the identification element, the micro-fluidic chip made of PMMA material has no adsorption influence on the target object, and the nonspecific adsorption is weaker; the micro-fluidic chip has stable property and can be stored for a long time; in addition, the micro-fluidic chip has the advantages of easily available processing materials and low cost. Therefore, the molecular imprinting electrostatic spinning fiber membrane is combined with the microfluidic technology, and the obtained microfluidic chip can be used for simply, rapidly and sensitively detecting the zearalenone, and is low in cost.
Drawings
Fig. 1 is a schematic perspective view of a microfluidic chip according to the present invention;
fig. 2 is a schematic plan view of a microfluidic chip according to the present invention;
FIG. 3 is an electron microscope image of the zearalenone molecularly imprinted polymer prepared in example 1;
FIG. 4 is a SEM scanning image of a fiber film obtained by electrospinning a polyacrylonitrile spinning solution with a concentration of 10% (w/w) in example 1;
FIG. 5 is an SEM scanning image of a molecularly imprinted electrospun fiber membrane of example 1;
FIG. 6 is a physical diagram of the microfluidic chip prepared in example 1;
fig. 7 is a physical diagram of a microfluidic chip connected to a teflon tube and a unidirectional constant flow pump prepared in example 1;
in FIGS. 1 and 2, 1-PMMA chip substrate, 2-microfluidic channel, 2-1-sample injection hole, 2-2-sample outlet hole, 3-pillow reaction tank, 4-fiber membrane.
Detailed Description
The invention provides a molecularly imprinted electrostatic spinning fiber membrane, which comprises a polyacrylonitrile nano electrostatic spinning fiber membrane and a zearalenone molecularly imprinted polymer dispersed in the fibers of the polyacrylonitrile nano electrostatic spinning fiber membrane. In the invention, the mass content of the zearalenone molecularly imprinted polymer in the molecularly imprinted electrostatic spinning fiber membrane is preferably 2-5%; the fiber diameter of the molecular imprinting electrostatic spinning fiber membrane is preferably 279-311 nm. The molecular imprinting electrostatic spinning fiber membrane provided by the invention has the characteristics of specific recognition of a molecular imprinting material, high specific surface area and sensitivity of an electrostatic nanofiber membrane, and not only can the zearalenone be specifically recognized, but also the cost is low and the stability is good.
The invention provides a preparation method of the molecular imprinting electrostatic spinning fiber membrane, which comprises the following steps:
mixing a zearalenone molecularly imprinted polymer and a polyacrylonitrile spinning solution to obtain a molecularly imprinted electrostatic spinning solution;
and carrying out electrostatic spinning on the molecular imprinting electrostatic spinning solution to obtain the molecular imprinting electrostatic spinning fiber membrane.
In the invention, polyacrylonitrile and N, N-dimethylformamide are preferably mixed to obtain the polyacrylonitrile spinning solution. In the present invention, the average molecular weight of the polyacrylonitrile is preferably 150000; the mixing of the polyacrylonitrile and the N, N-dimethylformamide is preferably carried out at room temperature, and the method of mixing the polyacrylonitrile and the N, N-dimethylformamide is not particularly limited, and the polyacrylonitrile may be sufficiently dissolved by a mixing method well known to those skilled in the art, specifically, stirring and mixing. In the present invention, the mass concentration of the polyacrylonitrile spinning solution is preferably 8 to 12%, more preferably 10%.
In the invention, the preparation method of the zearalenone molecularly imprinted polymer preferably comprises the following steps:
mixing warfarin sodium, acetonitrile and N-vinyl pyrrolidone for prepolymerization to obtain a prepolymerized product;
mixing the pre-polymerized product, trimethylolpropane trimethacrylate and azodiisobutyronitrile, and carrying out polymerization reaction at 60 ℃ to obtain a polymerized product;
eluting the polymerization product to obtain the zearalenone molecularly imprinted polymer.
In the present invention, the warfarin sodium (chemical name 3- (alpha-warfarin) -4-hydroxycoumarin sodium salt) is used as a virtual template; the acetonitrile is used as a pore-forming agent; the N-vinyl pyrrolidone is used as a functional monomer. In the invention, the dosage ratio of warfarin sodium to acetonitrile is preferably 0.1652g:50mL; the prepolymerization is preferably carried out at room temperature, and the time of the prepolymerization is preferably 2 to 3 hours. In the invention, acetonitrile is preferably added into warfarin sodium to dissolve warfarin sodium, and then N-vinyl pyrrolidone is added into the obtained solution to be mixed for prepolymerization. In the present invention, the trimethylolpropane trimethacrylate is used as a crosslinking agent; the azodiisobutyronitrile is used as an initiator; the molar ratio of the warfarin sodium to the N-vinyl pyrrolidone to the trimethylolpropane trimethacrylate is 1:4:10; the mass of the azodiisobutyronitrile is 1% of that of the acetonitrile. In the present invention, the polymerization time is preferably 24 hours; in the invention, preferably, trimethylolpropane trimethacrylate is added into the prepolymerization product to be uniformly mixed, azodiisobutyronitrile is added into the prepolymerization product to sequentially carry out ultrasonic and nitrogen blowing, and then the obtained reaction system is subjected to polymerization reaction under the condition of water bath oscillation. In the invention, the eluting reagent is a mixed solvent of methanol and acetic acid, and the volume ratio of the methanol to the acetic acid in the mixed solvent is preferably 9:1; and removing the template warfarin sodium through the elution. In the invention, the polymerization reaction has a predetermination, the warfarin sodium template molecule is a zearalenone structural analogue, and the polymer obtained by reacting the zearalenone structural analogue with a polymerization system such as a functional monomer and the like has a specific recognition capability on the structural analogue after eluting the template molecule, so that the prepared molecularly imprinted polymer can specifically recognize the zearalenone and is a zearalenone molecularly imprinted polymer.
In the invention, the dosage ratio of the zearalenone molecularly imprinted polymer to the polyacrylonitrile spinning solution is preferably 20-25 mg:1 to 2mL, more preferably 25mg:1mL. The preparation method comprises the steps of soaking the zearalenone molecularly imprinted polymer, adding the polyacrylonitrile spinning solution into the soaked zearalenone molecularly imprinted polymer, and stirring and mixing to obtain molecularly imprinted electrostatic spinning solution; the infiltration reagent is preferably N, N-dimethylformamide; the dosage of the reagent for infiltration is enough to fully infiltrate the zearalenone molecularly imprinted polymer.
After the molecular imprinting electrostatic spinning solution is obtained, the molecular imprinting electrostatic spinning solution is subjected to electrostatic spinning to obtain the molecular imprinting electrostatic spinning fiber membrane. The electrostatic spinning technology is that the electrostatic interaction is adopted to spin, under the action of stronger electrostatic force, high-voltage static electricity is formed between the spinning solution and the receiver, along with the increase of voltage, the surface tension of the spinning solution is gradually increased, and simultaneously, a Taylor cone is formed at a spinning nozzle, when the electric field force and the surface tension of liquid drops are balanced, the surface tension of the liquid is smaller than that of the electric field, the spinning solution is sprayed out in a thread shape and falls on the receiver, and in the process, the solvent is evaporated or melt is cooled and solidified, so that fibrous substances are obtained, and the formed nanofiber has the characteristics of lower cost, higher fiber yield and large specific surface area. In the invention, the spinning distance of the electrostatic spinning is preferably 10-15 cm, the spinning voltage is preferably 13-17 kV, and the flow rate is preferably 20-50 mu L/min; the spinning film obtained by electrostatic spinning is preferably received on aluminum foil paper, so that the molecular imprinting electrostatic spinning fiber film is obtained.
The preparation method of the molecular imprinting electrostatic spinning fiber membrane provided by the invention has the advantages of simple process, easiness in operation and convenience in realization of large-scale production.
The invention provides a microfluidic chip, which comprises a PMMA chip substrate 1, a microfluidic channel 2 arranged in the PMMA chip substrate 1 and a plurality of pillow-shaped reaction tanks 3 arranged on the microfluidic channel 2; the microfluidic channel 2 is divided into a plurality of sections by the pillow-shaped reaction tank;
each pillow-shaped reaction tank 3 is filled with a fiber membrane 4; the fiber membrane 4 is the molecular imprinting electrostatic spinning fiber membrane described in the technical scheme or the molecular imprinting electrostatic spinning fiber membrane prepared by the preparation method described in the technical scheme;
sample injection holes 2-1 and sample discharge holes 2-2 are respectively arranged at two ports of the microfluidic channel 2, and the sample injection holes 2-1 and the sample discharge holes 2-2 penetrate through the upper surface of the PMMA chip substrate 1.
The three-dimensional structure schematic diagram and the planar structure schematic diagram of the microfluidic chip provided by the invention are respectively shown in fig. 1 and fig. 2.
In the present invention, the microfluidic chip is preferably 5cm long and 3cm wide.
The microfluidic chip provided by the invention comprises a PMMA (polymethyl methacrylate) chip substrate 1. In the invention, the PMMA chip substrate has mechanical rigidity, is used as a carrier of the molecular imprinting electrostatic spinning fiber membrane, and can solve the problems of strong electrostatic adsorption force, fragile membrane body and difficult fixation of the molecular imprinting electrostatic spinning fiber membrane; in addition, the molecular engram polymer is used as an identification element, the micro-fluidic chip made of PMMA material has no adsorption influence on a target object, and the nonspecific adsorption is weak; the micro-fluidic chip made of PMMA material has stable property and can be stored for a long time; in addition, the PMMA chip substrate also has good chemical transparency, and raw materials are easy to obtain and low in cost.
The microfluidic chip provided by the invention comprises a microfluidic channel 2 arranged in a PMMA chip substrate 1, wherein the width of the microfluidic channel 2 is preferably 1mm, and the depth is preferably 2mm. In the invention, the microfluidic channel 2 is divided into a plurality of sections by a pillow-shaped reaction tank, and the microfluidic channel 2 is distributed in a shape like a Chinese character 'ji' in a chip substrate; in the invention, two ends of the micro-flow channel 2 are respectively provided with a sample inlet hole 2-1 and a sample outlet hole 2-2, and the sample inlet hole 2-1 and the sample outlet hole 2-2 penetrate through the upper surface of the PMMA chip substrate 1; the radius of the sample inlet hole 2-1 and the radius of the sample outlet hole 2-2 are preferably 0.6mm. In the invention, the sample injection hole 2-1 is preferably externally connected with a sample injection pipe, and the sample injection pipe is preferably a transparent Teflon pipe with the radius of 0.6 mm; the other end of the sample inlet pipe is connected with a unidirectional constant flow pump, the unidirectional constant flow pump is not particularly required, and the unidirectional constant flow pump well known by the person skilled in the art is adopted. In the present invention, the sample outlet 2-2 is preferably further externally connected with a collecting device, and the collecting device is preferably a collecting bottle. In the invention, the channel is a flowing channel of liquid to be tested.
The microfluidic chip provided by the invention comprises a plurality of pillow-shaped reaction tanks 3 arranged on the microfluidic channel 2. In the invention, the plurality of pillow-shaped reaction tanks 3 are preferably arranged in parallel; the length of the single pillow-shaped reaction tank 5 is preferably 10mm, the width is preferably 5mm, and the depth is preferably 2mm; the quantity of pillow shape reaction tank is 4 preferably, adsorbs step by step, is favorable to promoting adsorption efficiency. In the present invention, the pillow-shaped reaction tank may be simply referred to as a "pillow tank" for providing a reaction space. In the invention, each pillow-shaped reaction tank is internally provided with a fiber membrane 4; the fiber membrane 4 is the molecular imprinting electrostatic spinning fiber membrane described in the technical scheme or the molecular imprinting electrostatic spinning fiber membrane prepared by the preparation method described in the technical scheme; the length of the fiber membrane 4 is preferably 10mm, and the width is preferably 5mm, that is, the length and width dimensions of the fiber membrane are preferably the same as those of the pillow-shaped reaction tank.
According to the invention, the molecular imprinting electrostatic spinning fiber membrane is combined with the microfluidic technology, and the obtained microfluidic chip can be used for simply, rapidly and sensitively detecting the zearalenone, and is low in cost.
In the invention, the preparation method of the microfluidic chip preferably comprises the following steps: designing a microfluidic chip by utilizing AutoCAD software according to the technical scheme; engraving the PMMA chip substrate by adopting an engraving machine according to the design; filling the fiber membrane according to the technical scheme in each carved pillow-shaped reaction tank, and then packaging the carved whole concave surface. In the present invention, the parameter settings of the engraving machine are preferably: the diameter of the milling cutter is 0.8-1 mm, the row spacing is 0.5-1.0 mm, the spindle rotation speed is 2400rpm/min, and the feed rate is 200-400 mm/min. The method for filling the fiber membrane and packaging is not particularly required, in the embodiment of the invention, the cut fiber membrane is stuck on the acrylic adhesive tape, and then one surface of the acrylic adhesive tape, on which the fiber membrane is stuck, is correspondingly stuck on the whole engraved concave surface, so that the filling of the fiber membrane and the packaging of the chip are completed. In the invention, the packaging surface is the lower surface of the microfluidic chip. In the invention, the preparation method of the microfluidic chip is simple and easy to operate.
The invention provides a method for preprocessing a sample containing zearalenone, which comprises the following steps:
leading sample liquid to be detected into a microfluidic chip through a sample inlet hole 2-1, flowing through each pillow-shaped reaction tank 3 through a microfluidic channel 2, adsorbing gibberellin in the sample liquid step by step through a fiber membrane 4 in each pillow-shaped reaction tank 3, and flowing out through a sample outlet hole 2-2;
after the adsorption is finished, introducing an eluting reagent into the microfluidic chip through the sample inlet holes 2-1, flowing through each pillow-shaped reaction tank 3 through the microfluidic channel 2, gradually desorbing the fiber membrane 4 in each pillow-shaped reaction tank 3, flowing out eluent through the sample outlet holes 2-2, and collecting the eluent; the eluting reagent is methanol.
The method can detect the food and agricultural product samples containing the zearalenone, has no special requirement on the method for preparing the food and agricultural product samples into the sample liquid to be detected, and can specifically refer to the extraction step specified in GB 5009.209-2016 determination of zearalenone in food safety national standard food. In the invention, the sample injection hole 2-1 is preferably connected with a unidirectional constant flow pump by the transparent Teflon tube for injecting the sample liquid to be detected or the eluting reagent; the flow rate of the sample liquid to be detected is preferably 600-800 mu L/min; the flow rate of the eluent is preferably 500-1000 mu L/min; the adsorbed sample solution or eluent is preferably collected in a collection bottle.
The invention also provides a method for detecting zearalenone, which comprises the step of carrying out liquid phase mass spectrometry detection on the eluent in the technical scheme to obtain quantitative data of zearalenone. The method and the conditions for detecting the liquid phase mass spectrum have no special requirements, and the method and the conditions which are well known to the person skilled in the art can be adopted; quantitative analysis was performed by the liquid phase mass spectrometry detection. According to the invention, the microfluidic chip is used for carrying out specific enrichment on the zearalenone, and the liquid phase mass spectrum detection with high combination accuracy, strong sensitivity and mature technology can be used for carrying out simple, rapid and sensitive detection on the zearalenone.
The molecular imprinting electrostatic spinning fiber membrane, the preparation method thereof, the microfluidic chip and the method for detecting zearalenone provided by the invention are described in detail below with reference to examples, but are not to be construed as limiting the scope of the invention.
Example 1
Instrument: an electronic balance (Beijing Sidoris instruments Co., ltd.) was used to weigh the medicines, a 78-1 magnetic heating stirrer (nabobism instruments Co., ltd.), a unidirectional syringe pump TYD01-01-CE (Baoding Lei Fu fluid technology Co., ltd.), an API 5000 tandem quadrupole mass spectrometer (Agilent Co., ltd.) of a crystal grinding instrument F-engraving machine (Dongguan crystal grinding instrument technology Co., ltd.), and a Scanning Electron Microscope (SEM).
Raw materials: polyacrylonitrile (PAN) average molecular weight 150000 (beijing belvedere technologies), N-Dimethylformamide (DMF) (Sigma-Aldrich trade company, shanghai), polymethyl methacrylate (PMMA) 200cm x 200cm, acrylic tape.
Preparation of zearalenone molecularly imprinted polymer material:
weighing 0.1652g of the virtual template warfarin sodium, adding 50mL of a pore-forming agent acetonitrile into a round-bottom flask to dissolve the template, adding 213.73 mu L of a functional monomer N-vinyl pyrrolidone (NVP), uniformly mixing, and prepolymerizing for 2.5h; adding a crosslinking agent trimethylolpropane Trimethacrylate (TRIM) 1596.23 mu L into the prepolymerization reaction system, uniformly mixing, adding an initiator Azodiisobutyronitrile (AIBN) 50mg, performing ultrasonic nitrogen blowing for 10min, placing the round-bottom flask into a water bath vibration kettle, and reacting at 60 ℃ for 24h to obtain a polymerization product; and eluting the polymerization product to obtain the zearalenone molecularly imprinted polymer material, wherein the eluting reagent is a mixed solvent of methanol and acetic acid (the volume ratio is 9:1). FIG. 3 is an electron microscope image of the prepared zearalenone molecularly imprinted polymer.
Preparing a molecular imprinting electrostatic spinning fiber membrane:
weighing Polyacrylonitrile (PAN) particles with average molecular weight of 150000 into a 25mL brown bottle, adding N, N-Dimethylformamide (DMF) solution, placing a magneton, placing the magneton in a magnetic stirrer for dissolution at room temperature, and preparing a 10% (w/w) polyacrylonitrile spinning solution; weighing a certain amount of prepared zearalenone molecularly imprinted polymer material, adding 100 mu L of DMF solution for infiltration, adding a certain amount of 10% (w/w) polyacrylonitrile spinning solution, and continuing magnetic stirring and uniformly mixing to form the molecularly imprinted electrostatic spinning solution with the concentration of 25 mg/mL. And (3) adjusting electrostatic spinning equipment, carrying out electrostatic spinning under the conditions of a spinning distance of 15cm, a spinning voltage of 17kV and a flow rate of 20 mu L/min, and collecting a spinning film on aluminum foil paper with proper specification for standby.
FIG. 4 is an SEM electron microscope scan of a fiber film obtained by electrospinning a 10% (w/w) polyacrylonitrile spinning solution, and it can be seen that the polyacrylonitrile fiber has relatively uniform thickness and good morphology, and the fiber diameter is about 279-311 nm.
Fig. 5 is an SEM electron microscope scan of a molecularly imprinted electrospun fiber membrane, and it can be seen that the molecularly imprinted polymer is relatively uniformly distributed in the fiber.
The specific adsorption performance of the molecular imprinting electrostatic spinning fiber membrane is tested: cutting 4 molecularly imprinted electrospun fiber membranes with the size of a pillow pool, respectively placing the molecularly imprinted electrospun fiber membranes in 2mL small centrifuge tubes, adding 1mL of zearalenone standard solution, placing the small centrifuge tubes on an oscillator, carrying out shake adsorption for 2min, centrifuging, taking the supernatant for mass spectrum detection, and calculating the adsorption efficiency. As a result, the adsorption rate of the molecular imprinting electrostatic spinning fiber membrane to the zearalenone is 75.56-94.50%, and the molecular imprinting electrostatic spinning fiber membrane has no adsorption effect on biotoxins such as aflatoxin, so that the molecular imprinting electrostatic spinning fiber membrane has high-efficiency specific adsorption and high sensitivity to the zearalenone.
Preparation of a microfluidic chip:
designing a microfluidic chip shown in fig. 1 and 2 by using AutoCAD software, wherein the size of the microfluidic chip is 3cm x 5cm; the channel is 1mm wide and 2mm deep; four pillow pools (pillow-shaped reaction pools) are designed in parallel at the middle part of the chip, and each pillow pool is 5mm x 10mm x 2mm in size and communicated with a channel and is used for placing a molecularly imprinted electrostatic spinning fiber membrane; the sample inlet hole and the sample outlet hole are circular permeation holes with the radius of 0.6 mm; carving PMMA by a cnc engraving machine, wherein the diameter of a milling cutter is 1mm, the row spacing is 0.5mm, the spindle rotating speed is 2400rpm, and the feeding rate is 200mm/min; cutting the molecularly imprinted electrostatic spinning fiber membrane into a size of 5mm or 10mm, then pasting the size of the size on an acrylic adhesive tape, and correspondingly pasting one surface of the acrylic adhesive tape pasted with the fiber membrane on the whole engraved concave surface to finish filling of the fiber membrane in a pillow pool and packaging of chips. The obtained microfluidic chip is shown in FIG. 6. The sample injection hole of the chip is connected with a unidirectional constant flow pump (the physical diagram is shown in figure 7) by a transparent Teflon tube with the radius of 0.6mm, and the sample injection hole is connected with a collecting bottle.
Example 2
The microfluidic chip of example 1 was used to detect zearalenone in cereals (corn flour, wheat flour):
sample matrix extraction: referring to GB 5009.209-2016, weighing 5g of grain sample, adding 50 mu L of zearalenone standard solution with concentration of 100ppm (simulating corn containing zearalenone toxin), standing for 0.5h, adding acetonitrile-water extract with volume ratio of 9:1, shaking for 2min to extract zearalenone, and quantitatively filtering with filter paper; taking 1mL of filtrate, adding 4mL of water, uniformly mixing, and passing through a glass fiber membrane until the filtrate is clear, wherein the filtrate is used as a sample liquid to be detected for standby.
Enriching and purifying: the 1mL syringe is used for taking out filtrate, the filtrate is arranged on a unidirectional constant flow pump, the front end of the syringe is connected with a chip sample injection hole through a transparent Teflon tube, the flow speed is regulated to 600-800 mu L/min, and liquid flows out from the sample injection hole.
The elution process comprises the following steps: taking a new injector, sucking 1mL of methanol solution, adjusting the flow rate to 700 mu L/min by a unidirectional constant flow pump consistent with the enrichment and purification process, eluting by a chip, collecting eluent, and detecting by an on-machine (liquid mass spectrometry) to finish quantitative analysis.
The microfluidic chip of example 1 was tested to have an enrichment adsorption efficiency of 87.15% and 84.49% for zearalenone in corn flour and wheat flour, respectively.
The embodiment shows that the molecularly imprinted electrospun fiber membrane provided by the invention can specifically identify zearalenone, has high sensitivity, and is simple to prepare and low in cost; according to the invention, the molecular imprinting electrostatic spinning fiber membrane is combined with the microfluidic technology, and the obtained microfluidic chip can be used for simply, rapidly and sensitively detecting zearalenone and can be used as a substitute of an immunoaffinity column.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (9)
1. A molecularly imprinted electrospun fiber membrane, comprising a polyacrylonitrile nano electrospun fiber membrane and a zearalenone molecularly imprinted polymer dispersed in the fibers of the polyacrylonitrile nano electrospun fiber membrane;
the preparation method of the zearalenone molecularly imprinted polymer comprises the following steps:
mixing warfarin sodium, acetonitrile and N-vinyl pyrrolidone for prepolymerization to obtain a prepolymerized product; the prepolymerization reaction is carried out at room temperature, and the time of the prepolymerization reaction is 2-3 h;
mixing the pre-polymerized product, trimethylolpropane trimethacrylate and azodiisobutyronitrile, and carrying out polymerization reaction at 60 ℃ to obtain a polymerized product; the polymerization reaction time is 24 hours;
eluting the polymerization product to obtain a zearalenone molecularly imprinted polymer;
the molar ratio of the warfarin sodium to the N-vinyl pyrrolidone to the trimethylolpropane trimethacrylate is 1:4:10; the eluting reagent is a mixed solvent of methanol and acetic acid.
2. The molecularly imprinted electrospun fiber membrane according to claim 1, wherein the mass content of the zearalenone molecularly imprinted polymer in the molecularly imprinted electrospun fiber membrane is 2-5%; the fiber diameter of the molecular imprinting electrostatic spinning fiber membrane is 279-311 nm.
3. The method for preparing the molecularly imprinted electrospun fiber membrane according to claim 1 or 2, comprising the following steps:
mixing a zearalenone molecularly imprinted polymer and a polyacrylonitrile spinning solution to obtain a molecularly imprinted electrostatic spinning solution; the polyacrylonitrile spinning solution is obtained by mixing polyacrylonitrile and N, N-dimethylformamide, wherein the average molecular weight of the polyacrylonitrile is 150000, and the mass concentration of the polyacrylonitrile spinning solution is 8-12%;
and carrying out electrostatic spinning on the molecular imprinting electrostatic spinning solution to obtain the molecular imprinting electrostatic spinning fiber membrane.
4. The preparation method of claim 3, wherein the dosage ratio of the zearalenone molecularly imprinted polymer to the polyacrylonitrile spinning solution is 20-25 mg: 1-2 mL.
5. The method according to claim 3, wherein the electrostatic spinning distance is 10-15 cm, the spinning voltage is 13-17 kV, and the flow rate is 20-50. Mu.L/min.
6. The microfluidic chip is characterized by comprising a PMMA chip substrate (1), a microfluidic channel (2) arranged in the PMMA chip substrate (1) and a plurality of pillow-shaped reaction tanks (3) arranged on the microfluidic channel (2), wherein the microfluidic channel (2) is divided into a plurality of sections by the pillow-shaped reaction tanks;
each pillow-shaped reaction tank (3) is filled with a fiber membrane (4); the fiber membrane (4) is a molecular imprinting electrostatic spinning fiber membrane according to any one of claims 1-2 or a molecular imprinting electrostatic spinning fiber membrane prepared by the preparation method according to any one of claims 3-5;
sample injection holes (2-1) and sample discharge holes (2-2) are respectively arranged at two ports of the microfluidic channel (2), and the sample injection holes (2-1) and the sample discharge holes (2-2) penetrate through the upper surface of the PMMA chip substrate (1).
7. The microfluidic chip according to claim 6, wherein the microfluidic chip is 5cm long and 3cm wide; the width of the microfluidic channel (2) is 1mm, and the depth is 2mm; the length of each single pillow-shaped reaction tank (3) is 10mm, the width is 5mm, the depth is 2mm, and the number of the pillow-shaped reaction tanks (3) is 4; the length of the fiber membrane (4) is 10mm, and the width is 5mm; the radiuses of the sample inlet hole (2-1) and the sample outlet hole (2-2) are both 0.6mm.
8. A method for pretreatment of zearalenone-containing samples, comprising the steps of:
leading sample liquid to be detected into a microfluidic chip through a sample inlet (2-1), flowing through each pillow-shaped reaction tank (3) through a microfluidic channel (2), adsorbing gibberellin in the sample liquid step by step through a fiber membrane (4) in each pillow-shaped reaction tank (3), and flowing out through a sample outlet (2-2);
after the adsorption is finished, introducing an eluting reagent into the microfluidic chip through a sample inlet (2-1), flowing through each pillow-shaped reaction tank (3) through a microfluidic channel (2), gradually desorbing fiber membranes (4) in each pillow-shaped reaction tank (3), flowing out eluent through a sample outlet (2-2), and collecting the eluent; the eluting reagent is methanol; the microfluidic chip is the microfluidic chip according to claim 6 or 7.
9. A method for detecting zearalenone, which is characterized in that the eluent obtained by the method of claim 8 is subjected to liquid phase mass spectrometry detection to obtain quantitative data of zearalenone.
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