CN104007104B - Quickly detect kit and the using method thereof of organophosphorus pesticide - Google Patents
Quickly detect kit and the using method thereof of organophosphorus pesticide Download PDFInfo
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- CN104007104B CN104007104B CN201410219923.6A CN201410219923A CN104007104B CN 104007104 B CN104007104 B CN 104007104B CN 201410219923 A CN201410219923 A CN 201410219923A CN 104007104 B CN104007104 B CN 104007104B
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Abstract
The present invention relates to kit and the using method thereof of a kind of quick detection organophosphorus pesticide；Described kit includes: C enzyme bottle, T enzyme bottle, C substrate reactions cup, T substrate reactions cup, and described C enzyme bottle, T enzyme bottle are all with enzyme bottle seal cap sealing, and equipped with the lyophilized three enzyme intermixtures of enzyme；Described C substrate reactions cup, T substrate reactions cup are equipped with the lyophilized adsorptive pads containing acetylcholine, luminol and dimethylbenzidine.The present invention also relates to the method applying aforesaid kit quickly to detect organophosphorus pesticide.The present invention utilizes acetylcholinesterase, choline oxidase and peroxidase for catalyst system and catalyzing, and with luminol, the product after acecoline hydrolysis oxidation is produced chemiluminescence effect.Set up and test the method for organophosphorus pesticide in agricultural product delicately.The present invention has a tool detection, and speed is fast, operating process simple, stability and reproducible, need not the advantages such as expensive instrument, be very suitable for carrying out the field monitoring of organophosphorus pesticide in the market of farm produce.
Description
Technical field
The present invention relates to a kind of method of organophosphorus pesticide in quick detection food, be specifically related to a kind of quickly detection
The kit of organophosphorus pesticide and using method thereof.
Background technology
Residues of pesticides in agricultural product, food and medicine be still the healthy important latency of danger side of body human body it
One.In the agricultural production process of China, agricultural chemicals year usage amount still occupies first place in the world, and the making of organophosphorus pesticide
Consumption accounts for more than the 70% of whole pesticide dosage.According to document announcement in recent years, Chinese Provincial metropolitan agricultural trade city
The veterinary antibiotics organophosphorus pesticide disqualification rate that in Chang, the first quarter is sold accounts for more than 14%, in particular from
The persticide residue of greenhouse gardening vegetables is higher.Agricultural chemicals sampling monitoring finds, acephatemet, metrifonate, oxidation
Rogor, DDVP recall rate higher.The method of current detection organophosphorus pesticide mainly has: 1. spectroscopic methodology, mainly
Including AAS, chemoluminescence method, infrared and fluorescent spectrometry；2. red, orange, green, blue, yellow (ROGBY), mainly includes thin layer
Red, orange, green, blue, yellow (ROGBY), gas chromatography, liquid chromatography, supercritical fluid chromatography and Chromatography/Mass Spectrometry GC-MS；
3. immunological analysis method, mainly include enzyme linked immunosorbent assay analysis method, nm of gold immunity test strip method, immunity glimmering
Optical analysis etc.；4. sensor method, mainly includes piezocrystal hexavalent chrome bio-removal, optical fiber type enzyme sensor, has
Machine phosphorus hydrolase biology sensor, cholinesterase sensor and biomimetic sensor based on molecular imprinting.The fastest
Speed detection method, mainly includes quick measuring card method and tacheometer method.Spectroscopic methodology and chromatography be country authenticity detect agriculture
The standard method of organophosphorus pesticide in product, has high specificity, highly sensitive feature, and its weak point is to need
Utilize instrument costly and special tester, and operating process is considerably complicated, be not suitable for on-the-spot big
Organophosphorus pesticide in amount detection agricultural product.Immunological detection method highly sensitive and specific the strongest, but only
Can detect single or several organophosphorus pesticides, and the research and development time length of corresponding antibodies and cost is high.Immunology detection side
The further drawback of method is its repeatability and less stable.Sensor, method is the novel detection grown up in recent years
Technology, wherein about utilizing the report in terms of the current mode of cholinesterase and Copper diethlydithiocarbamate the most.Sensing skill
The feature of art is sensitive, simple to operate, but achievement in research is still at laboratory stage, there are still repeatability and stablizes
Property difference defect.Existing quick measuring card technology is to utilize the cholinesterase can the analog 2,6-dichloro of hydrolyse acetylcholine
Indophenols acetic acid esters and generate the 2 of blueness, 6-dichloroindophenol acetic acid, thus be determined with according to the change of quick measuring card color
The residual condition of machine phosphorus insecticide.Its advantage is that detection speed is fast, simple to operate.But the method is still the most qualitative
Method, sensitivity is low, and the detection limit for height of some species of organophosphor and carbamate pesticide is gone out veterinary antibiotics
National limit standard, additionally the organic pigment in sample can severe jamming measure and cause false negative result.And show
Some tacheometers are still a kind of portable spectrophotometer, need to coordinate complex detection reagent, and existing
Field is difficult to implement.
Finding by prior art documents, number of patent application is 2005100741791.3, entitled " change
Learn luminous organophosphorus pesticide residual analyzer and detection method thereof ".Its key technology is, at potassium peroxydisulfate and dioxy
Under conditions of changing the existence of titanium nanoparticle, ultraviolet catalytic effect is utilized to make the organophosphorus pesticide on vegetables or fruit turn
Turn to inorganic phosphate, then form phosphorus molybdenum vanadium heteropolyacid with molybdic acid and vanadate, produce after this material oxidation luminol
Biochemical luminescence, and be measured with Chemiluminescence Apparatus.But its defect is that reaction system is easily by extraneous phosphatic dry
Disturb, and make result that false negative to occur.Additionally the defect of this detection method be repeatability, stability the most not ideal enough,
The needs of qualitative and quantitative detection can not be met.
Summary of the invention
Present invention aims to the defect of prior art, it is provided that a kind of quick detection organophosphorus pesticide
Kit and using method thereof.
The present invention is to be realized by following technical scheme:
First aspect, the present invention relates to the kit of a kind of quick detection organophosphorus pesticide, and described kit includes:
C enzyme bottle, T enzyme bottle, C substrate reactions cup, T substrate reactions cup, described C enzyme bottle, T enzyme bottle all seal with enzyme bottle
Lid seals, and the three enzyme intermixtures being lyophilized equipped with enzyme；Described C substrate reactions cup, T substrate reactions cup are respectively provided with and contain
There is the lyophilized adsorptive pads of acecoline, luminol and dimethylbenzidine.
Second aspect, the invention still further relates to the method that aforesaid kit quickly detects organophosphorus pesticide, including following
Step:
1st step: the preparation of enzyme cryoprotector: sucrose, bovine serum albumin(BSA), glycine, glycerine are added calcium chloride
In solution, stirring is to being completely dissolved, and freezen protective is stand-by；
2nd step: the preparation of reaction enzymes liquid: described enzyme cryoprotector is diluted respectively acetylcholinesterase, choline oxidation
Enzyme, peroxidase, mixing, after freeze-drying, it is respectively charged in C enzyme bottle and T enzyme bottle；
Described cholinesterase is the acetylcholinesterase of commercialization, and its source can be from insect, animal's liver, fish
The acetylcholinesterase that brain, plant or technique for gene engineering produce, the acetylcholine ester in the application preferred insect source
Enzyme.Described choline oxidase is also the choline oxidase of commercialization, and it derives from Alcaligenes, soil Arthrobacter globiformis
Or the choline oxidase that technique for gene engineering produces, the choline oxidase in the application preferred Alcaligenes source.Described mistake
Oxide enzyme is commercially available horseradish peroxidase.
3rd step: the preparation of substrate reactions cup: reaction substrate pad is put into luminol, acecoline, dimethyl
In benzidine mixed solution, immersion drains, and after freeze-drying, puts into igelite reaction cup, be labeled as at the bottom of C
Thing reaction cup, T substrate reactions cup；
4th step: the test of sample: drip pure water respectively in C, T enzyme bottle, after shaking up, with plastic dropper to institute
State dropping measuring samples solution in T enzyme bottle, in C enzyme bottle, drip pure water or phosphate buffer, shake up, stand,
From described C, T enzyme bottle, draw product with dropper, be added drop-wise to respectively in C, T substrate reactions cup, at 2～8 points
On portable chemical light-emitting appearance, the luminous intensity of C, T substrate reactions cup, relatively pressing down according to luminous intensity is measured in clock
Rate processed, determines the concentration of organophosphor.
Preferably, in the 1st step, enzyme cryoprotector includes each component of following content: 5～15mmol/L calcium chloride
Solution 100 milliliters, sucrose 1～3 grams, glycine 1～3 grams, glycerine 5～15 milliliters, bovine serum albumin(BSA) 1～3
Gram.
Preferably, in the 2nd step, described acetylcholinesterase dilutes 3000～8000 times, and described choline oxidase is dilute
Releasing 200～800 times, described peroxidase dilutes 200～800 times.
Preferably, in the 3rd step, described C substrate reactions cup, the bottom surface internal diameter of T substrate reactions cup are 8～20mm, highly
It is 10～25mm.
Preferably, in the 3rd step, the thickness of described reaction substrate pad is 3～6mm.
Preferably, in the 3rd step, the concentration of described luminol is 1.0 × 10-4～3.0 × 10-4Mol/L, acetyl chloride
The concentration of choline is 3.0 × 10-3～15.0 × 10-3Mol/L, the concentration of dimethylbenzidine are 2.0 × 10-3～10.0
×10-3mol/L。
Preferably, in the 4th step, described in C, T enzyme bottle drip pure water be 50～200 microlitres, described in treat sample
Product solution is 50～200 microlitres, and in described C enzyme bottle, pure water is 50～200 microlitres, and phosphate buffer is 0.1
Mol/L, and pH is 7.0.
Product 100～300 microlitre preferably, in the 4th step, in described C, T substrate reactions cup.
The present invention has a following beneficial effect:
(1) detection sensitivity is high: the present invention has selected containing acetylcholinesterase, choline oxidase and peroxidating
Three enzyme systems of thing enzyme, and establish the chemiluminescence based on sensitizer and luminol, its sensitivity exceeds number
Ten times and thousands of times, and the detection range of linearity is the most broadening.The present invention detects the lowest detection of DDVP and metrifonate
Limit reaches 0.078ppm；The LDL of detection fenamiphos reaches 0.035ppm；Detection to carbosulfan limits
Reach 0.039ppm；
(2) kit of the present invention is easy to use, simple to operate, need not the reagent preparation process of complexity in testing,
Layman just can be readily accomplished detection according to kit specification；
(3) detection is quickly: the present invention can complete pretreatment process and the detection process of sample within half an hour, right
Veterinary antibiotics sample pre-treatments has only to 3～5 minutes, needs 10～25 minutes with kit test process.
(4) detectable organophosphor scope is wide: the present invention can detect include Aldicarb, flolimat, Rogor,
Orthene, sevin, metrifonate, Furadan, carbosulfan, basudin, DDVP, Carbicron,
Cadusafos, ethyl parathion, fenamiphos, malathion, acephatemet, parathion-methyl, Azodrin, oxamyl,
Phosmet, phoxim, chlopyrifos, isocarbophos, methyl different sulphur phosphorus, azinphos-methyl etc. have at interior several tens kinds
Machine phosphorus insecticide.The present invention also can detect that some heavy metals include that the materials such as cadmium, copper, lead, mercury and zinc are vegetables simultaneously
Residual on dish, fruit.
Accompanying drawing explanation
The detailed description made non-limiting example with reference to the following drawings by reading, other of the present invention is special
Levy, purpose and advantage will become more apparent upon:
Fig. 1 is the ideograph that the present invention quickly detects the kit of organophosphorus pesticide；
Wherein, 1 is containing comparison and three enzyme reagent bottle groups of test, including C enzyme bottle, T enzyme bottle；2 is enzyme bottle
Sealing lid, 3 is three enzyme reagent bottles；The 4 three enzyme intermixtures being lyophilized for enzyme；5 is substrate reactions cup cup；6 are
Lyophilized adsorptive pads containing acecoline, luminol and dimethylbenzidine；7 is comparison and test bigeminy
Substrate reactions cup group, including C substrate reactions cup, T substrate reactions cup.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in detail.Following example will assist in those skilled in the art
Member is further appreciated by the present invention, but limits the present invention the most in any form.It should be pointed out that, the common skill to this area
For art personnel, without departing from the inventive concept of the premise, it is also possible to make some deformation and improvement.These broadly fall into
Protection scope of the present invention.
Embodiment 1
The present embodiment relates to a kind of method utilizing kit quickly to detect organophosphorus pesticide, comprises the following steps:
The present embodiment detects the residual of organophosphorus pesticide in commercially available rape.
1, sample pre-treatments, by the weight commercially available rape leave within 10 grams 10 parts, is positioned over one with spiral
In the plastics of lid and glass container, it is also possible to conventional Cans.Add the pure water of about 10 milliliters, rock 3
After minute, after standing 2 minutes, take supernatant and can be directly used for test；
2, the preparation of enzyme cryoprotector: the compound method of the present embodiment is as follows: first preparation 5mmol/L calcium chloride
Solution 100 milliliters, is subsequently adding sucrose 1 gram, glycine 1 gram, glycerine 5 milliliters, bovine serum albumin(BSA) 1 gram,
Above composition is stirred while adding, until all of powdered reagent all dissolves, then cold under the conditions of-15 DEG C
Freeze preservation stand-by；
3, the preparation of reaction enzymes liquid: be diluted with above-mentioned enzyme cryoprotector by above-mentioned three kinds of enzyme liquid, will be every
The milligram acetylcholinesterase containing 425 international units of enzyme activity dilutes 3000 times, international containing 10 by every milligram
The acetylcholinesterase of unit of enzyme activity dilutes 200 times, by every milligram of horseradish containing 113 international units of enzyme activity
Peroxidase dilutes 200 times.Respectively take above-mentioned enzyme liquid 100 microlitre and be mixed in together, be divided in the peace of brown
In small jar bottle, it is placed on freeze drier after freeze-drying, seals after upper cover, be positioned in aluminium foil bag, room temperature preservation；
4, the preparation of substrate reactions cup: in the present embodiment, uses the igelite reaction cup, bottom surface internal diameter to be
8mm, height is 10mm, and the thickness of described reaction substrate pad is 3mm, and being put into by this pad containing concentration is 1.0
×10-4Mol/L luminol, 3.0 × 10-3Mol/L acecoline and 2.0 × 10-3Mol/L dimethyl joins
In the mixed solution of aniline, drain after soaking 3 minutes, and after freeze-drying, put in reaction cup, mark respectively
For C and T cup, enclosing in packaging bag, room temperature preservation is stand-by；
5, the test of sample: drip 50 microlitre pure water to C and T enzyme bottle respectively, after shaking up gently, to T enzyme
Drip 50 microlitre measuring samples solution with quantitative plastic dropper in Ping, drip with quantitative plastic dropper in C enzyme bottle
50 microlitre pure water, after shaking up gently, place 15 minutes, then draw from the enzyme bottle of above-mentioned C and T with dropper
Product 100 microlitre, is added drop-wise to respectively in C and T substrate reactions cup, sent out in portable chemical in 2 minutes
Measure the luminous intensity of C and T substrate reactions cup on light instrument, according to the relative inhibition of luminous intensity, determine organophosphor
Concentration.
Result shows, in 10 parts of tested rape samples, has the CL enhancement rate of 2 parts of rape samples respectively
It is 16% and 33%, it is determined that for the positive.The CL enhancement rate of remaining 8 parts of rape sample is respectively less than 15%.Comparison
Test positive is the flolimat of 5ppm.
Embodiment 2
The present embodiment relates to a kind of method utilizing kit quickly to detect organophosphorus pesticide, comprises the following steps:
The present embodiment detects the residual of organophosphorus pesticide in commercially available spinach.
1, sample pre-treatments, step is with the step 1 of embodiment 1；
2, the preparation of enzyme cryoprotector: the compound method of the present embodiment is as follows: first preparation 10mmol/L calcium chloride
Solution 100 milliliters, is subsequently adding sucrose 2 grams, glycine 2 grams, glycerine 10 milliliters, bovine serum albumin(BSA) 2
Gram.Above composition is stirred while adding, until all of powdered reagent all dissolves, then-25 DEG C of conditions
Lower freezen protective is stand-by；
3, the preparation of reaction enzymes liquid: be diluted with above-mentioned enzyme cryoprotector by above-mentioned three kinds of enzyme liquid, will be every
The milligram acetylcholinesterase containing 425 international units of enzyme activity dilutes 5000 times, international containing 10 by every milligram
The acetylcholinesterase of unit of enzyme activity dilutes 500 times, by every milligram of horseradish containing 113 international units of enzyme activity
Peroxidase dilutes 500 times.Respectively take above-mentioned enzyme liquid 200 microlitre and be mixed in together, be divided in the peace of brown
In small jar bottle, it is placed on freeze drier after freeze-drying, seals after upper cover, be positioned in aluminium foil bag, room temperature preservation.
4, the preparation of substrate reactions cup: in the present embodiment, uses polystyrene plastics reaction cup.Bottom surface internal diameter is
12mm, height is 15mm.The thickness of described reaction substrate pad is 4mm.Being put into by this pad containing concentration is 2.0
×10-4Mol/L luminol, 8.0 × 10-3Mol/L acecoline and 6.0 × 10-3Mol/L dimethyl joins
In the mixed solution of aniline, drain after soaking 3 minutes, and after freeze-drying, put in reaction cup, mark respectively
For C and T cup, enclosing in packaging bag, room temperature preservation is stand-by；
5, the test of sample: drip 100 microlitre pure water to C and T enzyme bottle respectively, after shaking up gently, to T
Enzyme bottle drips 100 microlitre measuring samples solution with quantitative plastic dropper.Drip with quantitative plastic dropper in C enzyme bottle
Add 100 microlitre pure water, after shaking up gently, place 20 minutes.Then with the dropper enzyme bottle from above-mentioned C and T
Draw product 200 microlitre, be added drop-wise to respectively in C and T substrate reactions cup.Then in portable in 5 minutes
The luminous intensity of C and T substrate reactions cup is measured, according to the relative inhibition of luminous intensity, really on formula Chemiluminescence Apparatus
Determine the concentration of organophosphor.
Result shows, in 10 parts of tested spinach samples, has the CL enhancement rate of 1 part of rape sample respectively
It is 21%, it is determined that for the positive.The CL enhancement rate of remaining 9 parts of spinach samples is respectively less than 15%.Comparison test
Positive is the Azodrin of 10ppm.
Embodiment 3
The present embodiment relates to a kind of method utilizing kit quickly to detect organophosphorus pesticide, comprises the following steps:
The present embodiment detects the residual of organophosphorus pesticide in commercially available Shuijing Pear.
1, sample pre-treatments: first Shuijing Pear is cut 1/8th sizes, is positioned in clean Cans.
Remaining step is with the step 1 of embodiment 1；
2, the preparation of enzyme cryoprotector: the compound method of the present embodiment is as follows: first preparation 15mmol/L calcium chloride
Solution 100 milliliters, is subsequently adding sucrose 3 grams, glycine 3 grams, glycerine 15 milliliters, bovine serum albumin(BSA) 3
Gram.Above composition is stirred while adding, until all of powdered reagent all dissolves, then-40 DEG C of conditions
Lower freezen protective is stand-by；
3, the preparation of reaction enzymes liquid: be diluted with above-mentioned enzyme cryoprotector by above-mentioned three kinds of enzyme liquid, will be every
The milligram acetylcholinesterase containing 425 international units of enzyme activity dilutes 8000 times, international containing 10 by every milligram
The acetylcholinesterase of unit of enzyme activity dilutes 800 times, by every milligram of horseradish containing 113 international units of enzyme activity
Peroxidase dilutes 800 times.Respectively take above-mentioned enzyme liquid 300 microlitre and be mixed in together, be divided in the peace of brown
In small jar bottle, it is placed on freeze drier after freeze-drying, seals after upper cover, be positioned in aluminium foil bag, room temperature preservation.
4, the preparation of substrate reactions cup: in the present embodiment, uses polycarbonate plastic reaction cup.Bottom surface internal diameter is
20mm, height is 25mm.When preparing reaction substrate pad.During described reaction substrate pad, thickness used
For 6mm.Then being put into by this pad containing concentration is 3.0 × 10-4Mol/L luminol, 15.0 × 10-3Mol/L chlorine
Change acetylcholine and 10.0 × 10-3In the mixed solution of mol/L dimethylbenzidine, drain after soaking 3 minutes,
And after freeze-drying, put in reaction cup, and it being respectively labeled as C and T cup, enclose in packaging bag, room temperature preservation is treated
With；
5, the test of sample: drip 200 microlitre pure water to C and T enzyme bottle respectively, after shaking up gently, to T
Enzyme bottle drips 200 microlitre measuring samples solution with quantitative plastic dropper.Drip with quantitative plastic dropper in C enzyme bottle
Add 200 microlitre pure water, after shaking up gently, place 25 minutes.Then with the dropper enzyme bottle from above-mentioned C and T
Draw product 300 microlitre, be added drop-wise to respectively in C and T substrate reactions cup.Then in portable in 8 minutes
The luminous intensity of C and T substrate reactions cup is measured, according to the relative inhibition of luminous intensity, really on formula Chemiluminescence Apparatus
Determine the concentration of organophosphor.
Result shows, the CL enhancement rate of 10 parts of tested Shuijing Pear samples is respectively less than 15%, and result is judged to all
For feminine gender.Comparison test positive is the acephatemet of 15ppm.
The present invention utilize acetylcholinesterase, choline oxidase and peroxidase for catalyst system and catalyzing, by acetyl chloride
Product after choline hydrolysis oxidation produces chemiluminescence effect with luminol.Due to organophosphorus pesticide can suppress this instead
Answer the activity of three kinds of enzymes of system, the especially activity of acetylcholinesterase, thus can set up and test agricultural production delicately
The method of organophosphorus pesticide in product, and utilize this principle to develop the chemiluminescence detection reagent of detection organophosphorus pesticide
Box.The present invention has a tool detection, and speed is fast, operating process simple, stability and reproducible, need not expensive instrument
Etc. advantage, it is very suitable for carrying out the field monitoring of organophosphorus pesticide in the market of farm produce.
Above the specific embodiment of the present invention is described.It is to be appreciated that the invention is not limited in above-mentioned spy
Determining embodiment, those skilled in the art can make various deformation or amendment within the scope of the claims, this not shadow
Ring the flesh and blood of the present invention.
Claims (9)
1. the kit of a quick detection organophosphorus pesticide, it is characterised in that described kit includes: C enzyme
Bottle, T enzyme bottle, C substrate reactions cup, T substrate reactions cup, described C enzyme bottle, T enzyme bottle all seal Gai Mi with enzyme bottle
Envelope, and the three enzyme intermixtures being lyophilized equipped with enzyme；Described C substrate reactions cup, T substrate reactions cup are respectively provided with containing chlorine
Change acetylcholine, luminol and the lyophilized adsorptive pads of dimethylbenzidine；
In the three enzyme intermixtures that described enzyme is lyophilized, the enzyme cryoprotector of employing is sucrose, bovine serum albumin(BSA), sweet ammonia
Acid, glycerine and the mixed solution of calcium chloride；Three enzymes are acetylcholinesterase, choline oxidase, peroxidase.
2. applying the method that the kit described in claim 1 quickly detects organophosphorus pesticide, its feature exists
In, comprise the following steps:
1st step: the preparation of enzyme cryoprotector: sucrose, bovine serum albumin(BSA), glycine, glycerine are added calcium chloride
In solution, stirring is to being completely dissolved, and freezen protective is stand-by；
2nd step: the preparation of reaction enzymes liquid: described enzyme cryoprotector is diluted respectively acetylcholinesterase, choline oxidation
Enzyme, peroxidase, mixing, after freeze-drying, it is respectively charged in C enzyme bottle and T enzyme bottle；
3rd step: the preparation of substrate reactions cup: reaction substrate pad is put into luminol, acecoline, dimethyl
In benzidine mixed solution, immersion drains, and after freeze-drying, puts into igelite reaction cup, be labeled as at the bottom of C
Thing reaction cup, T substrate reactions cup；
4th step: the test of sample: drip pure water respectively in C, T enzyme bottle, after shaking up, with plastic dropper to institute
State dropping measuring samples solution in T enzyme bottle, in C enzyme bottle, drip pure water or phosphate buffer, shake up, stand,
From described C, T enzyme bottle, draw product with dropper, be added drop-wise to respectively in C, T substrate reactions cup, at 2～8 points
On portable chemical light-emitting appearance, the luminous intensity of C, T substrate reactions cup, relatively pressing down according to luminous intensity is measured in clock
Rate processed, determines the concentration of organophosphor.
3. the method that kit as claimed in claim 2 quickly detects organophosphorus pesticide, it is characterised in that the 1st
In step, enzyme cryoprotector includes each component of following content: 5～15mmol/L calcium chloride solutions 100 milliliters, sugarcane
Sugar 1～3 gram, glycine 1～3 grams, glycerine 5～15 milliliters, bovine serum albumin(BSA) 1～3 grams.
4. the method that kit as claimed in claim 2 quickly detects organophosphorus pesticide, it is characterised in that the 2nd
In step, described acetylcholinesterase dilutes 3000～8000 times, and described choline oxidase dilutes 200～800 times,
Described peroxidase dilutes 200～800 times.
5. the method that kit as claimed in claim 2 quickly detects organophosphorus pesticide, it is characterised in that the 3rd
In step, described C substrate reactions cup, the bottom surface internal diameter of T substrate reactions cup are 8～20mm, and height is 10～25mm.
6. the method that kit as claimed in claim 2 quickly detects organophosphorus pesticide, it is characterised in that the
In 3 steps, the thickness of described reaction substrate pad is 3～6mm.
7. the method that kit as claimed in claim 2 quickly detects organophosphorus pesticide, it is characterised in that the
In 3 steps, the concentration of described luminol is 1.0 × 10-4～3.0 × 10-4Mol/L, the concentration of acecoline are
3.0×10-3～15.0 × 10-3Mol/L, the concentration of dimethylbenzidine are 2.0 × 10-3～10.0 × 10-3mol/L。
8. the method that kit as claimed in claim 2 quickly detects organophosphorus pesticide, it is characterised in that the 4th
In step, the described pure water that drips in C, T enzyme bottle is 50～200 microlitres, and described measuring samples solution is 50～200
Microlitre, in described C enzyme bottle, pure water is 50～200 microlitres, and phosphate buffer is 0.1mol/L, and pH
It is 7.0.
9. the method that kit as claimed in claim 2 quickly detects organophosphorus pesticide, it is characterised in that the 4th
Product 100～300 microlitre in step, in described C, T substrate reactions cup.
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| CN201410219923.6A CN104007104B (en) | 2014-05-22 | 2014-05-22 | Quickly detect kit and the using method thereof of organophosphorus pesticide |
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| CN201410219923.6A CN104007104B (en) | 2014-05-22 | 2014-05-22 | Quickly detect kit and the using method thereof of organophosphorus pesticide |
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| CN104007104B true CN104007104B (en) | 2016-09-07 |
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| CN105241882B (en) * | 2015-09-23 | 2017-10-27 | 山东大学 | Application of the Liquid Crystal Sensor in detection organophosphorus pesticide and carbamate chemicals for agriculture |
| CN105319211A (en) * | 2015-11-13 | 2016-02-10 | 无锡艾科瑞思产品设计与研究有限公司 | Kit for quickly detecting organophosphorus residues in agricultural products and application method of kit |
| CN105548431B (en) * | 2015-12-14 | 2016-09-07 | 山东省农业科学院植物保护研究所 | Detect the method for oxamyl and oxamyl oxime residual quantity in vegetables/fruit simultaneously |
| CN106191212B (en) * | 2016-08-08 | 2019-11-19 | 陈英就 | The preparation method of fast detecting reagent kit for toxicity of pesticide residue, the preparation method of stabilizer and cholinesterase enzyme powder |
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| CN114295769B (en) * | 2021-12-28 | 2023-09-19 | 云南省农业科学院质量标准与检测技术研究所 | Method for rapidly identifying recessive addition forbidden pesticides in fertilizer |
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| CN1773287A (en) * | 2005-09-30 | 2006-05-17 | 国家海洋局第一海洋研究所 | Organic phosphorus pesticide bionic identification test reagent kit |
| CN101378782A (en) * | 2005-12-21 | 2009-03-04 | 惠氏公司 | Protein formulations with reduced viscosity and uses thereof |
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| CN1773287A (en) * | 2005-09-30 | 2006-05-17 | 国家海洋局第一海洋研究所 | Organic phosphorus pesticide bionic identification test reagent kit |
| CN101378782A (en) * | 2005-12-21 | 2009-03-04 | 惠氏公司 | Protein formulations with reduced viscosity and uses thereof |
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| 农药残留快速检测-酶抑制法;程念政;《安徽化工》;20041231(第131期);第49页左栏第1-2段、右栏第2节 * |
| 环境和食品中农药残留快速检测技术研究进展;徐溢 等;《化学通报》;20061231;第69卷;第2页倒数第1-2段 * |
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