CN105319211A - Kit for quickly detecting organophosphorus residues in agricultural products and application method of kit - Google Patents
Kit for quickly detecting organophosphorus residues in agricultural products and application method of kit Download PDFInfo
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- CN105319211A CN105319211A CN201510776112.0A CN201510776112A CN105319211A CN 105319211 A CN105319211 A CN 105319211A CN 201510776112 A CN201510776112 A CN 201510776112A CN 105319211 A CN105319211 A CN 105319211A
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- 238000000034 method Methods 0.000 title claims description 17
- 102000004190 Enzymes Human genes 0.000 claims abstract description 140
- 108090000790 Enzymes Proteins 0.000 claims abstract description 140
- 238000010438 heat treatment Methods 0.000 claims abstract description 27
- 238000001514 detection method Methods 0.000 claims abstract description 26
- 238000012360 testing method Methods 0.000 claims abstract description 21
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical group ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims abstract description 16
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 claims abstract description 15
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229910052794 bromium Inorganic materials 0.000 claims abstract description 15
- IQFVPQOLBLOTPF-HKXUKFGYSA-L congo red Chemical compound [Na+].[Na+].C1=CC=CC2=C(N)C(/N=N/C3=CC=C(C=C3)C3=CC=C(C=C3)/N=N/C3=C(C4=CC=CC=C4C(=C3)S([O-])(=O)=O)N)=CC(S([O-])(=O)=O)=C21 IQFVPQOLBLOTPF-HKXUKFGYSA-L 0.000 claims abstract description 8
- 238000007789 sealing Methods 0.000 claims abstract description 8
- 229940088598 enzyme Drugs 0.000 claims description 135
- 239000000243 solution Substances 0.000 claims description 42
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 35
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 28
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 21
- 239000000047 product Substances 0.000 claims description 20
- 239000003987 organophosphate pesticide Substances 0.000 claims description 16
- 102000012440 Acetylcholinesterase Human genes 0.000 claims description 15
- 108010022752 Acetylcholinesterase Proteins 0.000 claims description 15
- 229940022698 acetylcholinesterase Drugs 0.000 claims description 15
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 14
- 108010000659 Choline oxidase Proteins 0.000 claims description 14
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 14
- 102000003992 Peroxidases Human genes 0.000 claims description 14
- 229930006000 Sucrose Natural products 0.000 claims description 14
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 14
- 229940098773 bovine serum albumin Drugs 0.000 claims description 14
- 238000004108 freeze drying Methods 0.000 claims description 14
- 235000011187 glycerol Nutrition 0.000 claims description 14
- 235000013905 glycine and its sodium salt Nutrition 0.000 claims description 14
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 14
- 239000008363 phosphate buffer Substances 0.000 claims description 14
- 239000005720 sucrose Substances 0.000 claims description 14
- 239000007795 chemical reaction product Substances 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 230000001681 protective effect Effects 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 238000010790 dilution Methods 0.000 claims description 6
- 239000012895 dilution Substances 0.000 claims description 6
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 3
- 239000003795 chemical substances by application Substances 0.000 abstract 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 2
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 239000000575 pesticide Substances 0.000 description 7
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- UFEJKYYYVXYMMS-UHFFFAOYSA-N methylcarbamic acid Chemical compound CNC(O)=O UFEJKYYYVXYMMS-UHFFFAOYSA-N 0.000 description 1
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kit for quickly detecting organophosphorus residues in agricultural products. The kit comprises a first color developing agent, a second color developing agent, a 20 ml heating test tube, a dropper, qualitative filter paper, a C enzyme bottle and a T enzyme bottle, wherein both the C enzyme bottle and the T enzyme bottle are sealed by enzyme bottle sealing covers and filled with a lyophilized three-enzyme mixture; the first color developing agent is a carbon tetrachloride solution with 4-8 ml of bromine; the second color developing agent is an ethanol solution with 4-10 g/L Congo red. The kit is low in cost, simple to manufacture and high in detection sensitivity, has the sensitivity which is tens and even thousands times higher than that in the prior art, is remarkably widened in detection linear range, convenient to use, and simple and quick in operation, does not need complicated reagent preparation processes during a test, and ensures that a non-specialized person can easily complete detection according to the specification of the kit.
Description
Technical field
The present invention relates to a kind of kit, specifically the kit of organophosphorus residue and using method thereof in a kind of quick detection agricultural product.
Background technology
Along with developing rapidly and strategical adjustment that the is agriculture and industrial structure of China's industry, the particularly change of planting type, original for secondary, even for the occurrence injury of the harmful organism such as Pest organism of quarantine is on the rise, the resistance to the action of a drug constantly rises, and causes pesticide dosage and spraying times constantly to increase.The phenomenon of the high poison of abuse, persistent pesticide remains incessant after repeated prohibition, and has and uses agricultural chemicals not according to code requirement, and what have even seriously violates major experiment standard.High-toxic pesticide is used to cause considerable part Practice for Pesticide Residue in Agricultural Products obviously to exceed standard, at people's cylinder accumulation after edible, diseases induced, sometimes even there is acute poisoning, seriously jeopardize people ' s health and life security.
Organophosphorus pesticide is wide in variety, drug effect is high, and purposes is wide, and easily decompose, generally do not accumulate in people, animal body, be a very important compounds in agricultural chemicals.But have many kinds very strong to the acute toxicity of people, animal, will take care especially in use, in recent years, the Cultivar development of high-efficiency low-toxicity is very fast, progressively instead of some high drugs kinds, makes the use of organophosphorus pesticide safer and more effective.But Pesticide Residue also produces to agricultural product and normal trade brings many negative effects.Along with the health perception of consumer, the enhancing day by day of environmental consciousness, people are concerned about in grain, veterinary antibiotics, Chinese crude drug and meat more with or without pollution by pesticides.The Ministry of Science and Technology set up food security key special subjects, also a large amount of funds of research and development has been dropped in food safety detection, Shuo Guo wholesale vegetable market, country fair, supermarket, hotel, school lunch service and personal user etc., have very large demand to the field quick detection of food security.
In current domestic detection vegetable and fruit, organophosphorus and N-methylcarbamate pesticides remains conventional method quick measuring card method and spectrophotometric method, wherein quick measuring card method needs to keep environment temperature 37 DEG C of conditions to react in the middle of testing process, testing result need be compared by naked eyes and blank card, for qualitatively judging, there is comparatively big error sometimes in the judgement of naked eyes, if measured result is positive, then need to adopt other analytical approachs to determine the content of its residues of pesticides further; Need in spectrophotometry process to keep environment temperature at about 37 DEG C, laboratory technician needs situ configuration substrate and the activity configuration acetylcholinesterase according to enzyme, testing result need calculate enzyme inhibition rate, if obtain positive result, need to determine residues of pesticides content further by other analytical approachs, this kind of method step is more loaded down with trivial details.
Summary of the invention
The kit of organophosphorus residue and using method thereof in the quick detection agricultural product that the object of the present invention is to provide a kind of structure simple, easy to use, to solve the problem proposed in above-mentioned background technology.
For achieving the above object, the invention provides following technical scheme:
The kit of organophosphorus residue in a kind of quick detection agricultural product, comprise the first developer, the second developer, 20mL heating test tube, dropper, qualitative filter paper, C enzyme bottle and T enzyme bottle, described C enzyme bottle and T enzyme bottle and all use enzyme bottle seal cap sealing, and three enzyme intermixtures of enzyme freeze-drying are housed, described first developer be 4-8mL the carbon tetrachloride solution of bromine, described second developer is the ethanolic solution that 4-10g/L is Congo red.
The using method of the kit of organophosphorus residue in described quick detection agricultural product, concrete steps are as follows:
(1) sucrose, bovine serum albumin(BSA), glycocoll, glycerine are added in calcium chloride solution, be stirred to and dissolve completely, freezen protective is stand-by, the concentration of described calcium chloride solution is 5-15mmol/L, the volume of calcium chloride solution is 100mL, and sucrose is 1-3g, and glycocoll is 1-3g, glycerine is 5-15mL, and bovine serum albumin(BSA) is 1-3g;
(2) described enzyme cryoprotector is diluted acetylcholinesterase, choline oxidase, peroxidase respectively, mixing, after freeze drying, be respectively charged in C enzyme bottle and T enzyme bottle, described acetylcholinesterase dilution 3000-8000 doubly, doubly, described peroxidase dilution 200-800 doubly for described choline oxidase dilution 200-800;
(3) the testing sample clear water that takes a morsel washes away surperficial earth, leaves standstill 3-5min after stirring 5-10min;
(4) respectively to C enzyme bottle, pure water is dripped in T enzyme bottle, after shaking up, in described T enzyme bottle, measuring samples solution is dripped with plastic dropper, pure water or phosphate buffer is dripped in C enzyme bottle, shake up, leave standstill, with dropper from described C enzyme bottle, reaction product is drawn in T enzyme bottle, drip respectively on qualitative filter paper, after volatilization is dry, the point sample of filter paper is disposed on the solution of the first developer of heating, described to C enzyme bottle, dripping pure water in T enzyme bottle is 50-200 μ L, described measuring samples solution is 50-200 μ L, in described C enzyme bottle, pure water is 50-200 μ L, phosphate buffer is 0.1mol/L, and pH is 7.0,
(5) after complete to bromine volatilization, the point sample place described in step 2 drips the second developer, observes color change;
(6) if the point sample place described in step (5) shows blueness, organophosphorus pesticide is remained; If the point sample place described in step (5) shows pink or redness, without organophosphorus pesticide.
As the present invention's further scheme: in described step (4), the temperature of the solution of heating the first developer is 45-55 DEG C.
Compared with prior art, the invention has the beneficial effects as follows:
Cost of the present invention is low, making is simple, detection sensitivity is high, its remolding sensitivity prior art exceeds decades of times and thousands of times, and it is significantly broadening to detect the range of linearity, easy to use, simple and quick, do not need complicated reagent preparation process in testing, layman just can complete detection easily according to kit instructions.
Embodiment
Be described in more detail below in conjunction with the technical scheme of embodiment to this patent.
Embodiment 1
The kit of organophosphorus residue in a kind of quick detection agricultural product, comprise the first developer, the second developer, 20mL heating test tube, dropper, qualitative filter paper, C enzyme bottle and T enzyme bottle, described C enzyme bottle and T enzyme bottle and all use enzyme bottle seal cap sealing, and three enzyme intermixtures of enzyme freeze-drying are housed, described first developer be 4mL the carbon tetrachloride solution of bromine, described second developer is the ethanolic solution that 4g/L is Congo red.
The using method of the kit of organophosphorus residue in described quick detection agricultural product, concrete steps are as follows:
(1) sucrose, bovine serum albumin(BSA), glycocoll, glycerine are added in calcium chloride solution, be stirred to and dissolve completely, freezen protective is stand-by, the concentration of described calcium chloride solution is 5mmol/L, the volume of calcium chloride solution is 100mL, and sucrose is 1g, and glycocoll is 1g, glycerine is 5mL, and bovine serum albumin(BSA) is 1g;
(2) described enzyme cryoprotector is diluted acetylcholinesterase, choline oxidase, peroxidase respectively, mixing, after freeze drying, be respectively charged in C enzyme bottle and T enzyme bottle, described acetylcholinesterase dilutes 3000 times, described choline oxidase dilutes 200 times, and described peroxidase dilutes 200 times;
(3) the testing sample clear water that takes a morsel washes away surperficial earth, leaves standstill 3min after stirring 5min;
(4) respectively to C enzyme bottle, pure water is dripped in T enzyme bottle, after shaking up, in described T enzyme bottle, measuring samples solution is dripped with plastic dropper, pure water or phosphate buffer is dripped in C enzyme bottle, shake up, leave standstill, with dropper from described C enzyme bottle, reaction product is drawn in T enzyme bottle, drip respectively on qualitative filter paper, after volatilization is dry, the point sample of filter paper is disposed on the solution of the first developer of heating, the temperature heating the solution of the first developer is 45 DEG C, heating process is ventilated, described to C enzyme bottle, dripping pure water in T enzyme bottle is 50 μ L, described measuring samples solution is 50 μ L, in described C enzyme bottle, pure water is 50 μ L, phosphate buffer is 0.1mol/L, and pH is 7.0,
(5) after complete to bromine volatilization, the point sample place described in step 2 drips the second developer, observes color change;
(6) if the point sample place described in step (5) shows blueness, organophosphorus pesticide is remained; If the point sample place described in step (5) shows pink or redness, without organophosphorus pesticide.
Embodiment 2
The kit of organophosphorus residue in a kind of quick detection agricultural product, comprise the first developer, the second developer, 20mL heating test tube, dropper, qualitative filter paper, C enzyme bottle and T enzyme bottle, described C enzyme bottle and T enzyme bottle and all use enzyme bottle seal cap sealing, and three enzyme intermixtures of enzyme freeze-drying are housed, described first developer be 5mL the carbon tetrachloride solution of bromine, described second developer is the ethanolic solution that 5g/L is Congo red.
The using method of the kit of organophosphorus residue in described quick detection agricultural product, concrete steps are as follows:
(1) sucrose, bovine serum albumin(BSA), glycocoll, glycerine are added in calcium chloride solution, be stirred to and dissolve completely, freezen protective is stand-by, the concentration of described calcium chloride solution is 8mmol/L, the volume of calcium chloride solution is 100mL, and sucrose is 1.5g, and glycocoll is 1.5g, glycerine is 8mL, and bovine serum albumin(BSA) is 1.5g;
(2) described enzyme cryoprotector is diluted acetylcholinesterase, choline oxidase, peroxidase respectively, mixing, after freeze drying, be respectively charged in C enzyme bottle and T enzyme bottle, described acetylcholinesterase dilutes 4000 times, described choline oxidase dilutes 300 times, and described peroxidase dilutes 300 times;
(3) the testing sample clear water that takes a morsel washes away surperficial earth, leaves standstill 3.5min after stirring 6min;
(4) respectively to C enzyme bottle, pure water is dripped in T enzyme bottle, after shaking up, in described T enzyme bottle, measuring samples solution is dripped with plastic dropper, pure water or phosphate buffer is dripped in C enzyme bottle, shake up, leave standstill, with dropper from described C enzyme bottle, reaction product is drawn in T enzyme bottle, drip respectively on qualitative filter paper, after volatilization is dry, the point sample of filter paper is disposed on the solution of the first developer of heating, the temperature heating the solution of the first developer is 48 DEG C, heating process is ventilated, described to C enzyme bottle, dripping pure water in T enzyme bottle is 80 μ L, described measuring samples solution is 80 μ L, in described C enzyme bottle, pure water is 80 μ L, phosphate buffer is 0.1mol/L, and pH is 7.0,
(5) after complete to bromine volatilization, the point sample place described in step 2 drips the second developer, observes color change;
(6) if the point sample place described in step (5) shows blueness, organophosphorus pesticide is remained; If the point sample place described in step (5) shows pink or redness, without organophosphorus pesticide.
Embodiment 3
The kit of organophosphorus residue in a kind of quick detection agricultural product, comprise the first developer, the second developer, 20mL heating test tube, dropper, qualitative filter paper, C enzyme bottle and T enzyme bottle, described C enzyme bottle and T enzyme bottle and all use enzyme bottle seal cap sealing, and three enzyme intermixtures of enzyme freeze-drying are housed, described first developer be 6mL the carbon tetrachloride solution of bromine, described second developer is the ethanolic solution that 6g/L is Congo red.
The using method of the kit of organophosphorus residue in described quick detection agricultural product, concrete steps are as follows:
(1) sucrose, bovine serum albumin(BSA), glycocoll, glycerine are added in calcium chloride solution, be stirred to and dissolve completely, freezen protective is stand-by, the concentration of described calcium chloride solution is 10mmol/L, the volume of calcium chloride solution is 100mL, and sucrose is 2g, and glycocoll is 2g, glycerine is 10mL, and bovine serum albumin(BSA) is 2g;
(2) described enzyme cryoprotector is diluted acetylcholinesterase, choline oxidase, peroxidase respectively, mixing, after freeze drying, be respectively charged in C enzyme bottle and T enzyme bottle, described acetylcholinesterase dilutes 5000 times, described choline oxidase dilutes 400 times, and described peroxidase dilutes 400 times;
(3) the testing sample clear water that takes a morsel washes away surperficial earth, leaves standstill 4min after stirring 7min;
(4) respectively to C enzyme bottle, pure water is dripped in T enzyme bottle, after shaking up, in described T enzyme bottle, measuring samples solution is dripped with plastic dropper, pure water or phosphate buffer is dripped in C enzyme bottle, shake up, leave standstill, with dropper from described C enzyme bottle, reaction product is drawn in T enzyme bottle, drip respectively on qualitative filter paper, after volatilization is dry, the point sample of filter paper is disposed on the solution of the first developer of heating, the temperature heating the solution of the first developer is 50 DEG C, heating process is ventilated, described to C enzyme bottle, dripping pure water in T enzyme bottle is 120 μ L, described measuring samples solution is 120 μ L, in described C enzyme bottle, pure water is 120 μ L, phosphate buffer is 0.1mol/L, and pH is 7.0,
(5) after complete to bromine volatilization, the point sample place described in step 2 drips the second developer, observes color change;
(6) if the point sample place described in step (5) shows blueness, organophosphorus pesticide is remained; If the point sample place described in step (5) shows pink or redness, without organophosphorus pesticide.
Embodiment 4
The kit of organophosphorus residue in a kind of quick detection agricultural product, comprise the first developer, the second developer, 20mL heating test tube, dropper, qualitative filter paper, C enzyme bottle and T enzyme bottle, described C enzyme bottle and T enzyme bottle and all use enzyme bottle seal cap sealing, and three enzyme intermixtures of enzyme freeze-drying are housed, described first developer be 7mL the carbon tetrachloride solution of bromine, described second developer is the ethanolic solution that 8g/L is Congo red.
The using method of the kit of organophosphorus residue in described quick detection agricultural product, concrete steps are as follows:
(1) sucrose, bovine serum albumin(BSA), glycocoll, glycerine are added in calcium chloride solution, be stirred to and dissolve completely, freezen protective is stand-by, the concentration of described calcium chloride solution is 12mmol/L, the volume of calcium chloride solution is 100mL, and sucrose is 2.5g, and glycocoll is 2.5g, glycerine is 12mL, and bovine serum albumin(BSA) is 2.5g;
(2) described enzyme cryoprotector is diluted acetylcholinesterase, choline oxidase, peroxidase respectively, mixing, after freeze drying, be respectively charged in C enzyme bottle and T enzyme bottle, described acetylcholinesterase dilutes 6000 times, described choline oxidase dilutes 600 times, and described peroxidase dilutes 600 times;
(3) the testing sample clear water that takes a morsel washes away surperficial earth, leaves standstill 4.5min after stirring 8min;
(4) respectively to C enzyme bottle, pure water is dripped in T enzyme bottle, after shaking up, in described T enzyme bottle, measuring samples solution is dripped with plastic dropper, pure water or phosphate buffer is dripped in C enzyme bottle, shake up, leave standstill, with dropper from described C enzyme bottle, reaction product is drawn in T enzyme bottle, drip respectively on qualitative filter paper, after volatilization is dry, the point sample of filter paper is disposed on the solution of the first developer of heating, the temperature heating the solution of the first developer is 52 DEG C, heating process is ventilated, described to C enzyme bottle, dripping pure water in T enzyme bottle is 150 μ L, described measuring samples solution is 150 μ L, in described C enzyme bottle, pure water is 150 μ L, phosphate buffer is 0.1mol/L, and pH is 7.0,
(5) after complete to bromine volatilization, the point sample place described in step 2 drips the second developer, observes color change;
(6) if the point sample place described in step (5) shows blueness, organophosphorus pesticide is remained; If the point sample place described in step (5) shows pink or redness, without organophosphorus pesticide.
Embodiment 5
The kit of organophosphorus residue in a kind of quick detection agricultural product, comprise the first developer, the second developer, 20mL heating test tube, dropper, qualitative filter paper, C enzyme bottle and T enzyme bottle, described C enzyme bottle and T enzyme bottle and all use enzyme bottle seal cap sealing, and three enzyme intermixtures of enzyme freeze-drying are housed, described first developer be 8mL the carbon tetrachloride solution of bromine, described second developer is the ethanolic solution that 10g/L is Congo red.
The using method of the kit of organophosphorus residue in described quick detection agricultural product, concrete steps are as follows:
(1) sucrose, bovine serum albumin(BSA), glycocoll, glycerine are added in calcium chloride solution, be stirred to and dissolve completely, freezen protective is stand-by, the concentration of described calcium chloride solution is 15mmol/L, the volume of calcium chloride solution is 100mL, and sucrose is 3g, and glycocoll is 3g, glycerine is 15mL, and bovine serum albumin(BSA) is 3g;
(2) described enzyme cryoprotector is diluted acetylcholinesterase, choline oxidase, peroxidase respectively, mixing, after freeze drying, be respectively charged in C enzyme bottle and T enzyme bottle, described acetylcholinesterase dilutes 8000 times, described choline oxidase dilutes 800 times, and described peroxidase dilutes 800 times;
(3) the testing sample clear water that takes a morsel washes away surperficial earth, leaves standstill 5min after stirring 10min;
(4) respectively to C enzyme bottle, pure water is dripped in T enzyme bottle, after shaking up, in described T enzyme bottle, measuring samples solution is dripped with plastic dropper, pure water or phosphate buffer is dripped in C enzyme bottle, shake up, leave standstill, with dropper from described C enzyme bottle, reaction product is drawn in T enzyme bottle, drip respectively on qualitative filter paper, after volatilization is dry, the point sample of filter paper is disposed on the solution of the first developer of heating, the temperature heating the solution of the first developer is 55 DEG C, heating process is ventilated, described to C enzyme bottle, dripping pure water in T enzyme bottle is 200 μ L, described measuring samples solution is 200 μ L, in described C enzyme bottle, pure water is 200 μ L, phosphate buffer is 0.1mol/L, and pH is 7.0,
(5) after complete to bromine volatilization, the point sample place described in step 2 drips the second developer, observes color change;
(6) if the point sample place described in step (5) shows blueness, organophosphorus pesticide is remained; If the point sample place described in step (5) shows pink or redness, without organophosphorus pesticide.
Cost of the present invention is low, making is simple, detection sensitivity is high, its remolding sensitivity prior art exceeds decades of times and thousands of times, and it is significantly broadening to detect the range of linearity, easy to use, simple and quick, do not need complicated reagent preparation process in testing, layman just can complete detection easily according to kit instructions.
Above the better embodiment of this patent is explained in detail, but this patent is not limited to above-mentioned embodiment, in the ken that one skilled in the relevant art possesses, various change can also be made under the prerequisite not departing from this patent aim.
Claims (3)
1. one kind is detected the kit of organophosphorus residue in agricultural product fast, it is characterized in that, comprise the first developer, the second developer, 20mL heating test tube, dropper, qualitative filter paper, C enzyme bottle and T enzyme bottle, described C enzyme bottle and T enzyme bottle and all use enzyme bottle seal cap sealing, and three enzyme intermixtures of enzyme freeze-drying are housed, described first developer be 4-8mL the carbon tetrachloride solution of bromine, described second developer is the ethanolic solution that 4-10g/L is Congo red.
2. the using method detecting the kit of organophosphorus residue in agricultural product fast as claimed in claim 1, it is characterized in that, concrete steps are as follows:
(1) sucrose, bovine serum albumin(BSA), glycocoll, glycerine are added in calcium chloride solution, be stirred to and dissolve completely, freezen protective is stand-by, the concentration of described calcium chloride solution is 5-15mmol/L, the volume of calcium chloride solution is 100mL, and sucrose is 1-3g, and glycocoll is 1-3g, glycerine is 5-15mL, and bovine serum albumin(BSA) is 1-3g;
(2) described enzyme cryoprotector is diluted acetylcholinesterase, choline oxidase, peroxidase respectively, mixing, after freeze drying, be respectively charged in C enzyme bottle and T enzyme bottle, described acetylcholinesterase dilution 3000-8000 doubly, doubly, described peroxidase dilution 200-800 doubly for described choline oxidase dilution 200-800;
(3) the testing sample clear water that takes a morsel washes away surperficial earth, leaves standstill 3-5min after stirring 5-10min;
(4) respectively to C enzyme bottle, pure water is dripped in T enzyme bottle, after shaking up, in described T enzyme bottle, measuring samples solution is dripped with plastic dropper, pure water or phosphate buffer is dripped in C enzyme bottle, shake up, leave standstill, with dropper from described C enzyme bottle, reaction product is drawn in T enzyme bottle, drip respectively on qualitative filter paper, after volatilization is dry, the point sample of filter paper is disposed on the solution of the first developer of heating, described to C enzyme bottle, dripping pure water in T enzyme bottle is 50-200 μ L, described measuring samples solution is 50-200 μ L, in described C enzyme bottle, pure water is 50-200 μ L, phosphate buffer is 0.1mol/L, and pH is 7.0,
(5) after complete to bromine volatilization, the point sample place described in step 2 drips the second developer, observes color change;
(6) if the point sample place described in step (5) shows blueness, organophosphorus pesticide is remained; If the point sample place described in step (5) shows pink or redness, without organophosphorus pesticide.
3. the using method of the kit of organophosphorus residue in quick detection agricultural product according to claim 2, is characterized in that, in described step (4), the temperature of the solution of heating the first developer is 45-55 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510776112.0A CN105319211A (en) | 2015-11-13 | 2015-11-13 | Kit for quickly detecting organophosphorus residues in agricultural products and application method of kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510776112.0A CN105319211A (en) | 2015-11-13 | 2015-11-13 | Kit for quickly detecting organophosphorus residues in agricultural products and application method of kit |
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| CN109540882A (en) * | 2018-12-20 | 2019-03-29 | 王红玲 | Organic phosphorus detection kit and its detection method in a kind of food |
| CN110923224A (en) * | 2019-11-27 | 2020-03-27 | 陕西金色科技有限公司 | Acetylcholinesterase protective agent, acetylcholinesterase solution and preparation method thereof |
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