CN101893574B - Kit for simultaneously detecting and tracing a plurality of environmental endocrine disruptors and application thereof - Google Patents
Kit for simultaneously detecting and tracing a plurality of environmental endocrine disruptors and application thereof Download PDFInfo
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Abstract
The invention relates to the field of environmental endocrine disruptors, and provides a kit for simultaneously detecting a plurality of environmental endocrine disruptors and application thereof. The kit is made by adopting the following steps: respectively preparing potassium permanganate solution, HCL solution and polysorbate-80 solution; uniformly mixing template molecules, a functional monomer, a cross-linking agent, a porogenic agent, an initiator and organic solvent in a certain molar rate to prepare molecularly imprinted sol-gel of the environmental endocrine disruptors; modifying the molecularly imprinted sol-gel on the inner surface of a microporous plate, sealing the inner surface by a preservative film for hours; soaking the modified microporous plate in eluant for eluting; and inserting ports of a sample injector of the microporous plate into luminescence reagent solution to obtain the kit for simultaneously detecting a plurality of environmental endocrine disruptors. The kit is connected to a multifunctional analyzer on the microporous plate to detect the environmental endocrine disruptors in an environmental sample extracting solution. The kit has the advantages of simple operation, less dosage of reagent, economy and practicality, low price and accessibility of chemical luminescent reagent, high sensitivity and simple operation.
Description
Technical field
The present invention relates to environment incretion interferent detection technique field; A kind of kit that can detect the multiple environment incretion interferent of trace simultaneously of more specifically saying so the invention still further relates to the application in the multiple environment incretion interferent in test sample of described kit.
Background technology
(Environmental endocrinedisrupting chemicals EDCs) is meant the xenobiotics of the processes such as synthetic, secretion, transportation, combination and metabolism that can disturb the endogenous hormones that keeps self balance in the biosome, regulate growth course to environment incretion interferent.Environment incretion interferent is divided into following several types: polychlorinated biphenyl, dioxin, induced by alkyl hydroxybenzene, dibutyl phthalate class, metal species, organochlorine compound environmental hormone class, plant estrogens, fungi estrogens extensively are present in the chemical industry articles for use such as herbicide, pesticide, germifuge, antiseptic, plastifier and washing agent.
EDCs has become the environmental contaminants of new generation that press for improvement at present, and the EDCs of trace can exert an influence to normal hormonal action, disturbs endocrine dysfunction, causes mammal and human dysgenesia, dysplasia and some pathologic damage.This type material does not exist only in industrial waste water, waste gas and the sanitary sewage, in agricultural product, possibly exist yet.Can not obtain in time, detect exactly if EDCs is residual, they just possibly get into body, and the directly or indirectly normal hormone metabolism of influence in vivo, bring serious harm to human health.
Set up the method for a kind of EDCs of polycomponent fast residue screening simultaneously, just become current this research field and need one of problem of solution badly.Present existing EDCs detects or the screening method mainly comprises in vivo studies, in vitro test, biomarker detection and instrument monitoring method etc.But these detections or screening method exist not enough:
1. uterus weight test and uterus calbindin CaBP-9k mRNA detection of expression and analysis of protein are often adopted in vivo studies, and this method reaction sensitivity has selectivity, but its cost is high, and sensitivity is low, is inappropriate for large-scale screening fast;
2. in vitro test mainly contains E-Screen method, receptor competition test etc., and the specificity of test itself is relatively poor, occurs false positive results easily;
3. the EDCs instrument detecting method of setting up by means of large-scale precision instruments such as HPLC or LC-MS coupling techniques, though the detection of EDCs is had very high sensitivity, this method can not once just identify the structure of analyte; Need confirm structure with GC-MS toward contact; Technical costs is higher, and required time of classic method is generally all longer, have in addition reach several hrs; Complicated operation can not be used for on-the-spot fast detecting.
4. above several method is for detection and the analysis of EDCs, have all generally that detection sensitivity is low, cost is high, as a result false positive, testing process complicated, be inappropriate for shortcoming such as field quick detection, therefore can not satisfy the needs of actual detected.
Summary of the invention
The present invention is in order to solve above technical matters, provides that a kind of detection speed is fast, selectivity is high, can detect the kit of multiple environment incretion interferent simultaneously.
The present invention also provides the application of a kind of this kit in detecting trace amount environment incretion jam object.
The present invention realizes in the following manner.
Detect the kit of the multiple environment incretion interferent of trace simultaneously, it is characterized in that: adopt following steps to process:
(1) select chemical illuminating reagent, prepare the solution of every kind of luminescence reagent respectively, said luminescence reagent is potassium permanganate, hydrochloric acid and Tween-80;
(2) selection function monomer, said function monomer can with environment incretion interferent synthetic molecules trace collosol and gel;
(3) selecting the environment incretion interferent molecule for use is template molecule, and template molecule, function monomer, crosslinking chemical, pore-foaming agent, initiating agent and organic solvent are mixed the molecular engram collosol and gel of processing environment incretion interferent by certain mol proportion;
(4) utilize modification technique layer by layer, the molecular engram collosol and gel is modified on the microwell plate inside surface, sealed up for safekeeping several hours with preservative film;
(5) modified microporous plate of drying in the step (4) is soaked in wash-out in the eluant, eluent, cleans, dry up;
(6) in the luminescence reagent solution that makes in the injector port difference inserting step (1) with the modified microporous plate that dries up in the step (5), obtain the said kit that detects the multiple environment incretion interferent of trace simultaneously.
The described kit that detects the multiple environment incretion interferent of trace simultaneously, its characteristic also is: said function monomer is acrylic acid, methacrylic acid or 4-vinylpridine; Said crosslinking chemical is trimethylol-propane trimethacrylate, N, N-methylene diacrylamine, 3,5-two (acrylic amide) benzoic acid, ethylene glycol dimethacrylate, divinylbenzene or pentaerythritol triacrylate; Said initiating agent is an azoisobutyronitrile; Said pore-foaming agent is methylene chloride, chloroform, acetonitrile, methyl alcohol, isopropyl alcohol, phenixin, N, N-dimethylformamide or dimethyl sulfoxide (DMSO); Said organic solvent is methylene chloride or phenixin.
The described kit that detects the multiple environment incretion interferent of trace simultaneously, its characteristic also is: the mol ratio of said template molecule, function monomer, crosslinking chemical, pore-foaming agent, initiating agent and organic solvent is 0.2～1: 2: 0.5～5: 40～65: 0.1～0.15: 2.0～30.
The described kit that detects the multiple environment incretion interferent of trace simultaneously, its characteristic also is: said molecular engram collosol and gel is modified on the microwell plate inside surface may further comprise the steps:
(a) microwell plate is used the NaOH ultrasonic cleaning, water cleans then, dries up;
(b) the molecular engram collosol and gel of multiple environment incretion interferent is modified in a certain order on the microwell plate inside surface.
The described kit that detects the multiple environment incretion interferent of trace simultaneously, its characteristic also is: said eluant, eluent is a methyl alcohol.
The described kit that detects the multiple environment incretion interferent of trace simultaneously, its characteristic also is: step (5) but in the molecular engram collosol and gel that the makes microwell plate room temperature preservation of modifying.
The described kit that detects the multiple environment incretion interferent of trace simultaneously, its characteristic also is: the microwell plate of modifying with the molecular engram collosol and gel in the step (4), sealed up for safekeeping 12 hours with preservative film.
Each described kit application in the testing environment incretion interferent that detects the multiple environment incretion interferent of trace simultaneously among the claim 1-7; It is characterized in that may further comprise the steps: the kit that will detect the multiple environment incretion interferent of trace when will make by above-mentioned any one method is connected to the microwell plate multi-functional analyzer, and the environment incretion interferent in the environmental sample extract is detected.
The invention has the beneficial effects as follows:
1, the molecular engram collosol and gel is modified in the method for microwell plate inside surface simple to operate, do not needed other materials just can be directly fixed on the microwell plate inside surface, avoided of the interference of other materials luminous detection.
2, the molecular engram collosol and gel is modified in the microwell plate inside surface, reagent use amount few (only need tens microlitres) can be implemented in when carrying out maximum 96 samples in ten minutes and measure.
3, the kit that detects multiple environment incretion interferent simultaneously provided by the present invention, economical and practical, chemical illuminating reagent is cheap and easy to get.
4, the high specificity that detects the kit of multiple environment incretion interferent simultaneously provided by the present invention, other non-specific molecules does not have influence to testing result in the sample; Highly sensitive, can reach the ng level.
5. the kit that detects multiple environment incretion interferent simultaneously provided by the present invention is simple to operation, does not need the technical professional just can operate.
Embodiment
Embodiment 1
Classes of herbicides: 2,4,5-trichloro biphenyl ethoxyacetic acid, 2,4-DCBP ethoxyacetic acid, Amrol, buckwheat remove Tianjin, alachlor, nitrofen, metribuzin
Detect the kit of 7 kinds of classes of herbicides environment incretion interferents of trace simultaneously, its preparation method may further comprise the steps:
(1) selects and to produce chemiluminescent luminescence reagent HCl, KMnO with the classes of herbicides environment incretion interferent
4, Tween-80, prepare HCl, KMnO respectively
4, Tween-80 solution, bottling, and on bottle, indicate A, B, C bottle and be beneficial to operating personnel and use;
(2) select can with the function monomer methacrylic acid (MAA) of classes of herbicides environment incretion interferent synthetic molecules trace collosol and gel;
(3) get 2,4,5-trichloro biphenyl ethoxyacetic acid, 2; 4-DCBP ethoxyacetic acid, Amrol, buckwheat remove Tianjin, alachlor, nitrofen, seven kinds of template molecules of metribuzin, respectively with function monomer methacrylic acid (MAA), and crosslinking chemical ethylene glycol dimethacrylate (EGDMA); The pore-foaming agent chloroform, the initiating agent azoisobutyronitrile, organic solvent dichloromethane is 0.2: 2: 0.5 in molar ratio: mix at 40: 0.1: 2.0; Obtain 2; 4,5-trichloro biphenyl ethoxyacetic acid, 2,4-DCBP ethoxyacetic acid, Amrol, buckwheat remove the molecular engram collosol and gel of Tianjin, alachlor, nitrofen, seven kinds of classes of herbicides environment incretion interferents of metribuzin;
(4) with used microwell plate with 1mol/L NaOH ultrasonic cleaning; Steam the thorough cleaning many times of water with secondary then, dry up, utilize modification technique layer by layer; With these 7 kinds of molecular engram collosol and gels according to metribuzin, nitrofen, 2; 4-DCBP ethoxyacetic acid, 2,4,5-trichloro biphenyl ethoxyacetic acid, alachlor, buckwheat go the order (the The English alphabet order of title spelling) of Tianjin, Amrol to modify the microwell plate inside surface from outside to inside.
(5) microwell plate of modifying was sealed up for safekeeping 12 hours with preservative film;
(6) modified microporous plate of drying in the step (5) was soaked in the methanol solution wash-out 120 minutes, thoroughly removes the template molecule in the collosol and gel, steam the thorough cleaning many times of water with secondary then, dry up.Can several such microwell plates of disposable making, room temperature just place can, waiting needs the time spent to bring use at any time.Among the luminescence reagent solution A that makes in the injector port difference inserting step (1) with microwell plate, B, the C, detected the kit of 7 kinds of classes of herbicides environment incretion interferents of trace simultaneously.
The kit that detects 7 kinds of environment incretion interferents of trace when making is connected to the microwell plate multi-functional analyzer; The injector port of microwell plate is to A, B, three kinds of luminescence reagent auto injections of C; Sample and A in this moment micropore, B, three kinds of reagent effects of C are luminous, and analyser records luminous value, changes the detection of next micropore then over to; And the like measure when realizing multiple environment incretion interferent, testing result is seen table 1.
Table 1 the present invention detects the kit testing result of 7 kinds of classes of herbicides environment incretion interferents of trace simultaneously
The result can find out from table 1: the kit that detects 7 kinds of classes of herbicides environment incretion interferents of trace simultaneously has the range of linearity of broad, higher sensitivity and lower detectability.
Embodiment 2
Insecticide: benzene hexachloride, parathion, carbaryl, Niran, hydroxyl Niran, D.D.T. (dichloro-diphenyl-trichloroethane), trichlorine acaricide, dieldrite, 5a,6,9,9a-hexahydro-6,9-methano-2,4, heptachlor, Heptachlor epoxide, malathion, Methomyl
Detect the kit of 13 kinds of insecticide environment incretion interferents of trace simultaneously, its preparation method may further comprise the steps:
(1) selects and to produce chemiluminescent luminescence reagent HCl, KMnO with the insecticide environment incretion interferent
4, Tween-80, prepare HCl, KMnO respectively
4, Tween-80 solution, bottling, and on bottle, indicate A, B, C bottle and be beneficial to operating personnel and use;
(2) select can with the function monomer acrylic acid of insecticide environment incretion interferent synthetic molecules trace collosol and gel;
(3) get benzene hexachloride, parathion, carbaryl, Niran, hydroxyl Niran, D.D.T. (dichloro-diphenyl-trichloroethane), trichlorine acaricide, dieldrite, 5a,6,9,9a-hexahydro-6,9-methano-2,4, heptachlor, Heptachlor epoxide, malathion, 13 kinds of template molecules of Methomyl; Respectively with function monomer acrylic acid (AA); Crosslinking chemical trimethylol-propane trimethacrylate (TRIM); The pore-foaming agent methylene chloride; The initiating agent azoisobutyronitrile; The organic solvent phenixin is 0.5: 2: 3 in molar ratio: mix at 45: 0.1: 2.2, obtain the molecular engram collosol and gel of benzene hexachloride, parathion, carbaryl, Niran, hydroxyl Niran, D.D.T. (dichloro-diphenyl-trichloroethane), trichlorine acaricide, dieldrite, 5a,6,9,9a-hexahydro-6,9-methano-2,4, heptachlor, Heptachlor epoxide, malathion, 13 kinds of insecticide environment incretion interferents of Methomyl;
(4) with used microwell plate with 1mol/L NaOH ultrasonic cleaning; Steam the thorough cleaning many times of water with secondary then; Dry up; Utilize modification technique layer by layer; 13 kinds of molecular engram collosol and gels that this is synthetic are modified the microwell plate inside surface from outside to inside according to 5a,6,9,9a-hexahydro-6,9-methano-2,4, Niran, heptachlor, D.D.T. (dichloro-diphenyl-trichloroethane), dieldrite, parathion, carbaryl, benzene hexachloride, Methomyl, Heptachlor epoxide, malathion, hydroxyl Niran, the acaricidal order of trichlorine (number of words from less to more, the The English alphabet order according to spelling that number of words is identical).
(5) microwell plate of modifying was sealed up for safekeeping 11 hours with preservative film;
(6) modified microporous plate of drying in the step (5) was soaked in the methanol solution wash-out 120 minutes, thoroughly removes the template molecule in the collosol and gel, steam the thorough cleaning many times of water with secondary then, dry up.Can several such microwell plates of disposable making, room temperature just place can, waiting needs the time spent to bring use at any time.Among the luminescence reagent solution A that makes in the injector port difference inserting step (1) with microwell plate, B, the C, detected the kit of 13 kinds of environment incretion interferents of trace simultaneously.
The kit that detects 13 kinds of environment incretion interferents of trace when making is connected to the microwell plate multi-functional analyzer; The injector port of microwell plate is to A, B, three kinds of luminescence reagent auto injections of C; Sample and A in this moment micropore, B, three kinds of reagent effects of C are luminous, and analyser records luminous value, changes the detection of next micropore then over to; And the like measure when realizing 13 kinds of environment incretion interferents, testing result is seen table 2.
Table 2 the present invention detects the kit testing result of 13 kinds of insecticide environment incretion interferents of trace simultaneously
The result can find out from table 2: the kit that detects 13 kinds of insecticide environment incretion interferents of trace simultaneously has the range of linearity of broad, higher sensitivity and lower detectability.
Embodiment 3
Germifuge class: hexachloro-benzene, Mancozeb, maneb, Carbatene, zineb, vinclozolin, ziram, benomyl
Detect the kit of 8 kinds of germifuge classes of trace environment incretion interferent simultaneously, its preparation method may further comprise the steps:
(1) selects and to produce chemiluminescent luminescence reagent HCl, KMnO with germifuge class environment incretion interferent
4, Tween-80, prepare HCl, KMnO respectively
4, Tween-80 solution, bottling, and on bottle, indicate A, B, C bottle and be beneficial to operating personnel and use;
(2) select can with the function monomer 4-vinylpridine of germifuge class environment incretion interferent synthetic molecules trace collosol and gel;
(3) get hexachloro-benzene, Mancozeb, maneb, Carbatene, zineb, vinclozolin, ziram, 8 kinds of template molecules of benomyl; Respectively with function monomer 4-vinylpridine (4-VP); Crosslinking chemical N, N-methylene diacrylamine; The pore-foaming agent acetonitrile; Initiating agent azoisobutyronitrile, organic solvent dichloromethane are 1: 2: 5 in molar ratio: mix at 65: 0.15: 30, obtain the molecular engram collosol and gel of 8 kinds of germifuge class environment incretion interferent hexachloro-benzenes, Mancozeb, maneb, Carbatene, zineb, vinclozolin, ziram, benomyl respectively;
(4) with used microwell plate with 1mol/L NaOH ultrasonic cleaning; Steam the thorough cleaning many times of water with secondary then; Dry up; Utilize modification technique layer by layer, 8 kinds of synthetic germifuge quasi-molecule trace collosol and gels are modified the microwell plate inside surface from outside to inside according to maneb, Carbatene, zineb, Mancozeb, ziram, benomyl, hexachloro-benzene order;
(5) microwell plate of modifying was sealed up for safekeeping 13 hours with preservative film;
(6) modified microporous plate of drying in the step (5) was soaked in the methanol solution wash-out 120 minutes, thoroughly removes the template molecule in the collosol and gel, steam the thorough cleaning many times of water with secondary then, dry up.Can several such microwell plates of disposable making, room temperature just place can, waiting needs the time spent to bring use at any time.Among the luminescence reagent solution A that makes in the injector port difference inserting step (1) with microwell plate, B, the C, detected the kit of 8 kinds of environment incretion interferents of trace simultaneously.
The kit that detects 8 kinds of environment incretion interferents of trace when making is connected to the microwell plate multi-functional analyzer; The injector port of microwell plate is to A, B, three kinds of luminescence reagent auto injections of C; Sample and A in this moment micropore, B, three kinds of reagent effects of C are luminous, and analyser records luminous value, changes the detection of next micropore then over to; And the like measure when realizing 8 kinds of environment incretion interferents, testing result is seen table 3.
Table 3 the present invention detects the kit testing result of 8 kinds of germifuge classes of trace environment incretion interferent simultaneously
The result can find out from table 3: the kit that detects 8 kinds of germifuge classes of trace environment incretion interferent simultaneously has the range of linearity of broad, higher sensitivity and lower detectability.
Embodiment 4
Antiseptic kind: pentachloro-phenol, tributyl tin, triphenyltin
Detect the kit of 3 kinds of antiseptic kind environment incretion interferents of trace simultaneously, its preparation method may further comprise the steps:
(1) selects and to produce chemiluminescent luminescence reagent HCl, KMnO with the antiseptic kind environment incretion interferent
4, Tween-80, prepare HCl, KMnO respectively
4, Tween-80 solution, bottling, and on bottle, indicate A, B, C bottle and be beneficial to operating personnel and use;
(2) select can with the function monomer 4-vinylpridine of antiseptic kind environment incretion interferent synthetic molecules trace collosol and gel;
(3) get pentachloro-phenol, tributyl tin, 3 kinds of template molecules of triphenyltin, respectively with the function monomer 4-vinylpridine, crosslinking chemical 3; 5-two (acrylic amide) benzoic acid; Pore-foaming agent methyl alcohol, the initiating agent azoisobutyronitrile, organic solvent dichloromethane is 0.5: 2: 3 in molar ratio: 45: 0.1: 4.0; Mix, obtain the molecular engram collosol and gel of pentachloro-phenol, tributyl tin, 3 kinds of germifuge classes of triphenyltin environment incretion interferent;
(4) with used microwell plate with 1mol/L NaOH ultrasonic cleaning; Steam the thorough cleaning many times of water with secondary then; Dry up; Utilize modification technique layer by layer, the molecular engram collosol and gel of 3 kinds of germifuge class environment incretion interferents is modified the microwell plate inside surface from outside to inside according to the order of triphenyltin, tributyl tin, pentachloro-phenol;
(5) microwell plate of modifying was sealed up for safekeeping 12 hours with preservative film;
(6) modified microporous plate of drying in the step (5) was soaked in the methanol solution wash-out 120 minutes, thoroughly removes the template molecule in the collosol and gel, steam the thorough cleaning many times of water with secondary then, dry up.Can several such microwell plates of disposable making, room temperature just place can, waiting needs the time spent to bring use at any time.Among the luminescence reagent solution A that makes in the injector port difference inserting step (1) with microwell plate, B, the C, detected the kit of 3 kinds of antiseptic kind environment incretion interferents of trace simultaneously.
The kit that detects 3 kinds of antiseptic kind environment incretion interferents of trace when making is connected to the microwell plate multi-functional analyzer; The injector port of microwell plate is to A, B, three kinds of luminescence reagent auto injections of C; Sample and A in this moment micropore, B, three kinds of reagent effects of C are luminous, and analyser records luminous value, changes the detection of next micropore then over to; And the like measure when realizing 3 kinds of antiseptic kind environment incretion interferents, testing result is seen table 4.
Table 4 the present invention detects the kit testing result of 3 kinds of antiseptic kind environment incretion interferents of trace simultaneously
The result can find out from table 4: the kit that detects 3 kinds of antiseptic kind environment incretion interferents of trace simultaneously has the range of linearity of broad, higher sensitivity and lower detectability.
Embodiment 5
The plastic plasticizer class: two (2-ethyl) the own esters of adjacent benzenetricarboxylic acid, benzyl phthalate, n-butyl phthalate, the two cyclohexyls of phthalic acid, the two diethylesters of phthalic acid, hexane diacid are two-2-Octyl Nitrite, dipropyl phthalate
Detect the kit of 7 kinds of plastic plasticizer classes of trace environment incretion interferent simultaneously, its preparation method may further comprise the steps:
(1) selects and to produce chemiluminescent luminescence reagent HCl, KMnO with the classes of herbicides environment incretion interferent
4, Tween-80, prepare HCl, KMnO respectively
4, Tween-80 solution, bottling, and on bottle, indicate A, B, C bottle and be beneficial to operating personnel and use;
(2) select can with the function monomer acrylic acid of plastic plasticizer class environment incretion interferent synthetic molecules trace collosol and gel;
(3) get two (2-ethyl) the own esters of adjacent benzenetricarboxylic acid, benzyl phthalate, n-butyl phthalate; The two cyclohexyls of phthalic acid, the two diethylesters of phthalic acid, hexane diacid are two-2-Octyl Nitrite, 7 kinds of template molecules of dipropyl phthalate, respectively with function monomer acrylic acid; Crosslinking chemical divinylbenzene (DVB), pore-foaming agent N, N-dimethylformamide; The initiating agent azoisobutyronitrile; Organic solvent dichloromethane is 0.7: 2: 3 in molar ratio: 60: 0.12: 15, mix, and obtain 7 kinds of plastic plasticizer class environment incretion jam object molecular imprinting collosol and gels;
(4) with used microwell plate with 1mol/L NaOH ultrasonic cleaning; Steam the thorough cleaning many times of water with secondary then; Dry up; Utilize modification technique layer by layer, with 7 kinds of plastic plasticizer class environment incretion jam object molecular imprinting collosol and gels according to hexane diacid two-2-Octyl Nitrite, dipropyl phthalate, n-butyl phthalate, the two diethylesters of phthalic acid, the two cyclohexyls of phthalic acid, two (2-ethyl) the own esters of adjacent benzenetricarboxylic acid, benzyl phthalate order modify the microwell plate inside surface from outside to inside;
(5) microwell plate of modifying was sealed up for safekeeping 12 hours with preservative film;
(6) modified microporous plate of drying in the step (5) was soaked in the methanol solution wash-out 120 minutes, thoroughly removes the template molecule in the collosol and gel, steam the thorough cleaning many times of water with secondary then, dry up.Can several such microwell plates of disposable making, room temperature just place can, waiting needs the time spent to bring use at any time.Among the luminescence reagent solution A that makes in the injector port difference inserting step (1) with microwell plate, B, the C, detected the kit of 7 kinds of plastic plasticizer classes of trace environment incretion interferent simultaneously.
The kit that detects 7 kinds of plastic plasticizer classes of trace environment incretion interferent when making is connected to the microwell plate multi-functional analyzer; The injector port of microwell plate is to A, B, three kinds of luminescence reagent auto injections of C; Sample and A in this moment micropore, B, three kinds of reagent effects of C are luminous, and analyser records luminous value, changes the detection of next micropore then over to; And the like measure when realizing 7 kinds of plastic plasticizer class environment incretion interferents, testing result is seen table 5.
Table 5 the present invention detects the kit testing result of 7 kinds of plastic plasticizer classes of trace environment incretion interferent simultaneously
The result can find out from table 5: the kit that detects 7 kinds of plastic plasticizer classes of trace environment incretion interferent simultaneously has the range of linearity of broad, higher sensitivity and lower detectability.
Embodiment 6
Washing agent class: C6 alkylphenol, nonyl phenol, 4-octyl phenol
Detect the kit of 3 kinds of washing agent classes of trace environment incretion interferent simultaneously, its preparation method may further comprise the steps:
(1) selects and to produce chemiluminescent luminescence reagent HCl, KMnO with washing agent class environment incretion interferent
4, Tween-80, prepare HCl, KMnO respectively
4, Tween-80 solution, bottling, and on bottle, indicate A, B, C bottle and be beneficial to operating personnel and use;
(2) select can with the function monomer methacrylic acid of washing agent class environment incretion interferent synthetic molecules trace collosol and gel;
(3) get C6 alkylphenol, nonyl phenol, 3 kinds of template molecules of 4-octyl phenol; Respectively with function monomer methacrylic acid (MAA); Crosslinking chemical pentaerythritol triacrylate (PETRA), pore-foaming agent dimethyl sulfoxide (DMSO), initiating agent azoisobutyronitrile; Organic solvent dichloromethane is 1: 2: 1 in molar ratio: mix at 55: 0.1: 15, obtain the molecular engram collosol and gel of 3 kinds of washing agent class environment incretion interferents;
(4) with used microwell plate with 1mol/L NaOH ultrasonic cleaning; Steam the thorough cleaning many times of water with secondary then; Dry up; Utilize modification technique layer by layer, 3 kinds of washing agent class environment incretion jam object molecular imprinting collosol and gels are modified the microwell plate inside surface in proper order according to C6 alkylphenol, 4-octyl phenol, nonyl phenol;
(5) microwell plate of modifying was sealed up for safekeeping 12 hours with preservative film;
(6) modified microporous plate of drying in the step (5) was soaked in the methanol solution wash-out 120 minutes, thoroughly removes the template molecule in the collosol and gel, steam the thorough cleaning many times of water with secondary then, dry up.Can several such microwell plates of disposable making, room temperature just place can, waiting needs the time spent to bring use at any time.Among the luminescence reagent solution A that makes in the injector port difference inserting step (1) with microwell plate, B, the C, detected the kit of 3 kinds of environment incretion interferents of trace simultaneously.
The kit that detects 3 kinds of environment incretion interferents of trace when making is connected to the microwell plate multi-functional analyzer; The injector port of microwell plate is to A, B, three kinds of luminescence reagent auto injections of C; Sample and A in this moment micropore, B, three kinds of reagent effects of C are luminous, and analyser records luminous value, changes the detection of next micropore then over to; And the like measure when realizing 3 kinds of environment incretion interferents, testing result is seen table 6.
Table 6 the present invention detects the kit testing result of 3 kinds of washing agent classes of trace environment incretion interferent simultaneously
The result can find out from table 6: the kit that detects 3 kinds of washing agent classes of trace environment incretion interferent simultaneously has the range of linearity of broad, higher sensitivity and lower detectability.
Claims (7)
1. kit that detects the multiple environment incretion interferent of trace simultaneously is characterized in that: adopt following steps to process:
(1) select chemical illuminating reagent, prepare the solution of every kind of luminescence reagent respectively, said luminescence reagent is potassium permanganate, hydrochloric acid and Tween-80;
(2) selection function monomer, said function monomer can with environment incretion interferent synthetic molecules trace collosol and gel;
(3) selecting the environment incretion interferent molecule for use is template molecule, and template molecule, function monomer, crosslinking chemical, pore-foaming agent, initiating agent and organic solvent are mixed the molecular engram collosol and gel of processing environment incretion interferent by certain mol proportion;
(4) utilize modification technique layer by layer, the molecular engram collosol and gel is modified on the microwell plate inside surface, sealed up for safekeeping several hours with preservative film;
(5) modified microporous plate of drying in the step (4) is soaked in wash-out in the eluant, eluent, cleans, dry up;
(6) in the luminescence reagent solution that makes in the injector port difference inserting step (1) with the modified microporous plate that dries up in the step (5), obtain the said kit that detects the multiple environment incretion interferent of trace simultaneously;
Wherein, in the step (4) said molecular engram collosol and gel modified on the microwell plate inside surface and may further comprise the steps:
(a) microwell plate is used the NaOH ultrasonic cleaning, water cleans then, dries up;
(b) the molecular engram collosol and gel of multiple environment incretion interferent is modified in a certain order on the microwell plate inside surface.
2. detect the kit of the multiple environment incretion interferent of trace according to time described in the claim 1, it is characterized in that: said function monomer is acrylic acid, methacrylic acid or 4-vinylpridine; Said crosslinking chemical is trimethylol-propane trimethacrylate, N, N-methylene diacrylamine, 3,5-two (acrylic amide) benzoic acid, ethylene glycol dimethacrylate, divinylbenzene or pentaerythritol triacrylate; Said initiating agent is an azoisobutyronitrile; Said pore-foaming agent is methylene chloride, chloroform, acetonitrile, methyl alcohol, isopropyl alcohol, phenixin, N, N-dimethylformamide or dimethyl sulfoxide (DMSO); Said organic solvent is methylene chloride or phenixin.
3. detect the kit of the multiple environment incretion interferent of trace according to time described in the claim 1, it is characterized in that: the mol ratio of said template molecule, function monomer, crosslinking chemical, pore-foaming agent, initiating agent and organic solvent is 0.2～1: 2: 0.5～5: 40～65: 0.1～0.15: 2.0～30.
4. detect the kit of the multiple environment incretion interferent of trace according to time described in the claim 1, it is characterized in that: said eluant, eluent is a methyl alcohol.
5. detect the kit of the multiple environment incretion interferent of trace according to described in the claim 1 time, it is characterized in that: step (5) but in the molecular engram collosol and gel that the makes microwell plate room temperature preservation of modifying.
6. detect the kit of the multiple environment incretion interferent of trace according to time described in the claim 1, it is characterized in that: the microwell plate of modifying with the molecular engram collosol and gel in the step (4), sealed up for safekeeping 12 hours with preservative film.
7. each described kit application in the testing environment incretion interferent that detects the multiple environment incretion interferent of trace simultaneously among the claim 1-6; It is characterized in that may further comprise the steps: the kit that will detect the multiple environment incretion interferent of trace when will make by above-mentioned any one method is connected to the microwell plate multi-functional analyzer, and the environment incretion interferent in the environmental sample extract is detected.
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| CN2010102102529A CN101893574B (en) | 2010-06-28 | 2010-06-28 | Kit for simultaneously detecting and tracing a plurality of environmental endocrine disruptors and application thereof |
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| CN2010102102529A CN101893574B (en) | 2010-06-28 | 2010-06-28 | Kit for simultaneously detecting and tracing a plurality of environmental endocrine disruptors and application thereof |
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Families Citing this family (3)
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| CN102372821A (en) * | 2010-08-27 | 2012-03-14 | 中国石油化工股份有限公司 | Dibutyl phthalate molecularly imprinted polymer (DBP-MIP) and preparation method thereof |
| CN104650289A (en) * | 2015-03-06 | 2015-05-27 | 天津科技大学 | Preparation method of diuron molecularly imprinted polymer and application of diuron molecularly imprinted polymer |
| CN113209948A (en) * | 2021-05-19 | 2021-08-06 | 齐鲁工业大学 | Preparation method and application of multi-template molecularly imprinted composite material |
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