CN109001195A - A kind of vegetable pesticide residue detection method - Google Patents
A kind of vegetable pesticide residue detection method Download PDFInfo
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- CN109001195A CN109001195A CN201811068491.8A CN201811068491A CN109001195A CN 109001195 A CN109001195 A CN 109001195A CN 201811068491 A CN201811068491 A CN 201811068491A CN 109001195 A CN109001195 A CN 109001195A
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- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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Abstract
The present invention provides a kind of vegetable pesticide residue detection method, and in particular to the Detection Technologies of Pesticide Residues field includes the following steps: that S1, reagent prepare and prepare: configuration buffer, butyrylcholine esterase liquid, substrate and color developing agent；S2, preparation of samples: blade or melon meat etc. being cut into small pieces and are put into extraction flask, and buffer is added, and concussion obtains extracting solution, static, and it is spare to obtain supernatant；S3, comparison liquid detection: it takes buffer and butyrylcholine esterase liquid to be added in reaction flask, adds color developing agent, placed in constant incubator certain time, substrate is added, pours into cuvette, is detected with the residual tacheometer device of agriculture；S4, analyte sample fluid detection: supernatant, butyrylcholine esterase liquid and color developing agent are taken, a period of time is placed in constant incubator, is poured into cuvette, shakes, is detected with the residual tacheometer device of agriculture；S5, testing result is calculated.The present invention, which has, improves preparation rate, and finding speed is fast, advantage easy to operate.
Description
Technical field
The invention belongs to the Detection Technologies of Pesticide Residues fields, and in particular to a kind of vegetable pesticide residue detection method.
Background technique
In China, the pesticide residue of vegetable and fruit is extremely serious.Fruit and vegetable excessive pesticide residues major reason is caused to be
Some peasant households fruit and vegetable plant in using country forbid using highly toxic pesticide, cause persticide residue excessive.Food utensil
There is the remaining fruit and vegetable of exceeded toxic pesticides, person poultry poisoning's event can be caused, great damage is caused to human body.Existing rank
Section, there are many kinds of China's Fast Determination of Pesticide Residue methods, is largely divided into biochemical assay and chromatographic detection two according to principle
Major class.It is accurate that chromatographic detection measures structure, but minute is long, and the enzyme inhibition rate method in biochemical assay is due to having speed
Degree is fast, is swift in response, is easy to operate, the features such as required infusion of financial resources is few, is listed in national proposed standard method, especially needle
It is measured to organic phosphorus with carbamate chemicals for agriculture, enzyme inhibition is a kind of quick detection technique, mainly to enzyme
The content of product is measured after hydrolysis substrate, and a kind of detection method that its measurement result is compared with standard items can be divided into
Potentiometric titration, manometric method, pH differential technique (△ pH) and 4 kinds of colorimetric method, but the measurement knot of present enzyme inhibition rate method
Structure accuracy rate is lower.
Therefore, it is badly in need of a kind of raising preparation rate, finding speed is fast, vegetable pesticide residue detection method easy to operate.
Summary of the invention
Measurement structure accuracy rate in order to solve the problems, such as existing enzyme inhibition rate method is lower, and the object of the present invention is to provide one
Kind vegetable pesticide residue detection method, with fast, the easy to operate advantage of finding speed.
The present invention provides the following technical solutions:
A kind of vegetable pesticide residue detection method, includes the following steps:
S1, reagent prepare and prepare: configuration buffer, butyrylcholine esterase liquid, substrate and color developing agent；
S2, preparation of samples: taking the vegetables or melon and fruit that surface is clean, and leaf vegetables takes blade-section, and fruit takes surface, by blade or
Melon meat etc., which is cut into small pieces, is put into extraction flask, and buffer is added, and concussion obtains extracting solution, static, and it is spare to obtain supernatant；
S3, comparison liquid detection: it takes buffer and butyrylcholine esterase liquid to be added in reaction flask, color developing agent is added, in perseverance
It is placed certain time in warm incubator, substrate is added, pours into cuvette, is detected with the residual tacheometer device of agriculture；
S4, analyte sample fluid detection: supernatant made from S2 step, butyrylcholine esterase liquid and color developing agent are taken, in constant temperature
A period of time is placed in incubator, is poured into cuvette, is shaken, is detected with the residual tacheometer device of agriculture；
S5, calculate testing result: testing result is calculated by following mathematical expression:
Wherein: the difference of light absorption value after △ Ac- control group 3min and before 3min, after △ As- sample 3min with extinction before 3min
The difference of value.
When inhibiting rate < 70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard is not contained in vegetables, works as inhibiting rate
When >=70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard may be contained in vegetables, the sample will have 2 times or more at this time
Repetition detection, detected through being repeated several times, < 70% reproducibility should then determine qualification 80% or more.
Preferably, the configuration of buffer is buffer dissolved in purified water in the S1 step；Butyrylcholine esterase liquid
Configuration is butyrylcholine esterase buffer solution；The configuration of substrate is substrate pulvis buffer solution；The configuration of color developing agent
It is color developing agent buffer solution.
Preferably, leaf vegetables weight/buffer dosage is 1g/20mL, the use of melon and fruit weight buffer in the S2 step
Amount is 2g/20mL.
Preferably, it is 1-3min, static 4-6min that the time is shaken in the S2 step.
Preferably, the temperature of constant incubator is 37-38 DEG C in the S3 step and the S4 step, standing time
35-45min。
The beneficial effects of the present invention are: the present invention is using the colorimetric method in enzyme inhibition, colorimetric method is changed by color developing agent
A kind of measuring method for becoming substrate hydrolysis coloured product, with fast, the easy to operate feature of finding speed, the present invention uses butyryl
The preparation rate of measurement result can be improved in cholinesterase.
Specific embodiment
Embodiment 1
A kind of vegetable pesticide residue detection method, includes the following steps:
S1, reagent prepare and prepare: the dissolved in purified water of 1 bag of buffer 1000mL when configuration buffer is stored at room temperature；
10mL buffer is added when configuring butyrylcholine esterase liquid in every bottle of butyrylcholine esterase liquid pulvis to be dissolved, 0-4 DEG C of preservation；
Configure every bottom of bottle medicinal powder 10mL buffer solution when substrate, 0-4 DEG C of preservation；Every bottle of color developing agent 10mL when configuring color developing agent
Buffer solution is protected from light stored refrigerated；
S2, preparation of samples: taking the vegetables that surface is clean, and leaf vegetables takes blade-section, 1g blade is accurately weighed with balance, by leaf
The fritter that piece is cut into 1cm square size is put into extraction flask, and 20mL buffer is added, and is shaken 1min, is obtained extracting solution, static
It is spare to obtain supernatant by 4min；
S3, comparison liquid detection: it takes buffer 3ml and 50ul butyrylcholine esterase liquid to be added in reaction flask, adds 50ul
Color developing agent, places 35min in 38 DEG C of constant incubators, and 50ul substrate is added, pours into cuvette, with the residual tacheometer device of agriculture into
Row detection, blank solution light absorption value control between 0.4-0.8；
S4, analyte sample fluid detection: supernatant made from S2 step, 50ul butyrylcholine esterase liquid and 50ul colour developing are taken
Agent is placed 35min in 38 DEG C of constant incubators, is poured into cuvette, and concussion is detected with the residual tacheometer device of agriculture；
S5, calculate testing result: testing result is calculated by following mathematical expression:
Wherein: the difference of light absorption value after △ Ac- control group 3min and before 3min, after △ As- sample 3min with extinction before 3min
The difference of value.
When inhibiting rate < 70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard is not contained in vegetables, works as inhibiting rate
When >=70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard may be contained in vegetables, the sample will have 2 times or more at this time
Repetition detection, detected through being repeated several times, < 70% reproducibility should then determine qualification 80% or more.
Embodiment 2
A kind of vegetable pesticide residue detection method, includes the following steps:
S1, reagent prepare and prepare: the dissolved in purified water of 1 bag of buffer 1000mL when configuration buffer is stored at room temperature；
10mL buffer is added when configuring butyrylcholine esterase liquid in every bottle of butyrylcholine esterase liquid pulvis to be dissolved, 0-4 DEG C of preservation；
Configure every bottom of bottle medicinal powder 10mL buffer solution when substrate, 0-4 DEG C of preservation；Every bottle of color developing agent 10mL when configuring color developing agent
Buffer solution is protected from light stored refrigerated；
S2, preparation of samples: taking the melon and fruit that surface is clean, and melon and fruit takes fruit parts, 2g melon and fruit is accurately weighed with balance, by melon
The fritter that fruit is cut into 1cm square size is put into extraction flask, and 20mL buffer is added, and is shaken 2min, is obtained extracting solution, static
It is spare to obtain supernatant by 5min；
S3, comparison liquid detection: it takes buffer 3ml and 50ul butyrylcholine esterase liquid to be added in reaction flask, adds 50ul
Color developing agent, places 40min in 37 DEG C of constant incubators, and 50ul substrate is added, pours into cuvette, with the residual tacheometer device of agriculture into
Row detection, blank solution light absorption value control between 0.4-0.8；
S4, analyte sample fluid detection: supernatant made from S2 step, 50ul butyrylcholine esterase liquid and 50ul colour developing are taken
Agent is placed 40min in 37 DEG C of constant incubators, is poured into cuvette, and concussion is detected with the residual tacheometer device of agriculture；
S5, calculate testing result: testing result is calculated by following mathematical expression:
Wherein: the difference of light absorption value after △ Ac- control group 3min and before 3min, after △ As- sample 3min with extinction before 3min
The difference of value.
When inhibiting rate < 70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard is not contained in vegetables, works as inhibiting rate
When >=70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard may be contained in vegetables, the sample will have 2 times or more at this time
Repetition detection, detected through being repeated several times, < 70% reproducibility should then determine qualification 80% or more.
Embodiment 3
A kind of vegetable pesticide residue detection method, includes the following steps:
S1, reagent prepare and prepare: the dissolved in purified water of 1 bag of buffer 1000mL when configuration buffer is stored at room temperature；
10mL buffer is added when configuring butyrylcholine esterase liquid in every bottle of butyrylcholine esterase liquid pulvis to be dissolved, 0-4 DEG C of preservation；
Configure every bottom of bottle medicinal powder 10mL buffer solution when substrate, 0-4 DEG C of preservation；Every bottle of color developing agent 10mL when configuring color developing agent
Buffer solution is protected from light stored refrigerated；
S2, preparation of samples: taking the vegetables that surface is clean, and leaf vegetables takes blade-section, 1g blade is accurately weighed with balance, by leaf
The fritter that piece is cut into 1cm square size is put into extraction flask, and 20mL buffer is added, and is shaken 3min, is obtained extracting solution, static
It is spare to obtain supernatant by 6min；
S3, comparison liquid detection: it takes buffer 3ml and 50ul butyrylcholine esterase liquid to be added in reaction flask, adds 50ul
Color developing agent places 45min in constant incubator, and 50ul substrate is added, pours into cuvette, is examined with the residual tacheometer device of agriculture
It surveys, blank solution light absorption value controls between 0.4-0.8；
S4, analyte sample fluid detection: supernatant made from S2 step, 50ul butyrylcholine esterase liquid and 50ul colour developing are taken
45min is put in agent in constant incubator, is poured into cuvette, and concussion is detected with the residual tacheometer device of agriculture；
S5, calculate testing result: testing result is calculated by following mathematical expression:
Wherein: the difference of light absorption value after △ Ac- control group 3min and before 3min, after △ As- sample 3min with extinction before 3min
The difference of value.
When inhibiting rate < 70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard is not contained in vegetables, works as inhibiting rate
When >=70%, certain organic phosphorus or carbamate chemicals for agriculture residual hazard may be contained in vegetables, the sample will have 2 times or more at this time
Repetition detection, detected through being repeated several times, < 70% reproducibility should then determine qualification 80% or more.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, although referring to aforementioned reality
Applying example, invention is explained in detail, for those skilled in the art, still can be to aforementioned each implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features.It is all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (5)
1. a kind of vegetable pesticide residue detection method, which comprises the steps of:
S1, reagent prepare and prepare: configuration buffer, butyrylcholine esterase liquid, substrate and color developing agent；
S2, preparation of samples: taking the vegetables or melon and fruit that surface is clean, and leaf vegetables takes blade-section, and fruit takes surface, by blade or melon meat
It is put into extraction flask etc. being cut into small pieces, buffer is added, concussion obtains extracting solution, and it is static, it is spare to obtain supernatant；
S3, comparison liquid detection: taking buffer and butyrylcholine esterase liquid to be added in reaction flask, add color developing agent, trains in constant temperature
It supports and is placed certain time in case, substrate is added, pours into cuvette, is detected with the residual tacheometer device of agriculture；
S4, analyte sample fluid detection: supernatant made from the S2 step, butyrylcholine esterase liquid and color developing agent are taken, in constant temperature
A period of time is placed in incubator, is poured into cuvette, is shaken, is detected with the residual tacheometer device of agriculture；
S5, calculate testing result: testing result is calculated by following mathematical expression:
Wherein: the difference of light absorption value after △ Ac- control group 3min and before 3min, after △ As- sample 3min with light absorption value before 3min it
Difference.
2. a kind of vegetable pesticide residue detection method according to claim 1, which is characterized in that buffered in the S1 step
The configuration of liquid is buffer dissolved in purified water；The configuration of butyrylcholine esterase liquid is butyrylcholine esterase buffer solution；
The configuration of substrate is substrate pulvis buffer solution；The configuration of color developing agent is color developing agent buffer solution.
3. a kind of vegetable pesticide residue detection method according to claim 1, which is characterized in that leaf vegetables in the S2 step
Weight/buffer dosage is 1g/20mL, and the dosage of melon and fruit weight buffer is 2g/20mL.
4. a kind of vegetable pesticide residue detection method according to claim 1, which is characterized in that shaken in the S2 step
Time is 1-3min, static 4-6min.
5. a kind of vegetable pesticide residue detection method according to claim 1, which is characterized in that the S3 step and described
The temperature of constant incubator is 37-38 DEG C in S4 step, standing time 35-45min.
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| CN201811068491.8A CN109001195A (en) | 2018-09-13 | 2018-09-13 | A kind of vegetable pesticide residue detection method |
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| CN201811068491.8A CN109001195A (en) | 2018-09-13 | 2018-09-13 | A kind of vegetable pesticide residue detection method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109406201A (en) * | 2018-12-18 | 2019-03-01 | 安徽中青检验检测有限公司 | A kind of food content of beary metal device for fast detecting |
| CN110208256A (en) * | 2019-06-14 | 2019-09-06 | 智锐达仪器科技南通有限公司 | A kind of Detecting Pesticide pulvis and preparation method, cuvette, detection method |
| CN112326646A (en) * | 2020-11-03 | 2021-02-05 | 童超 | Pesticide residue detection method for food detection |
| CN115656531A (en) * | 2022-12-08 | 2023-01-31 | 深圳市赛特罗生物医疗技术有限公司 | Integrated pesticide residue standardized rapid detection box and detection method |
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Patent Citations (5)
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| WO2001006249A2 (en) * | 1999-07-02 | 2001-01-25 | Conceptual Mindworks, Inc. | Organic semiconductor recognition complex and system |
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Cited By (5)
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| CN109406201A (en) * | 2018-12-18 | 2019-03-01 | 安徽中青检验检测有限公司 | A kind of food content of beary metal device for fast detecting |
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| CN110208256A (en) * | 2019-06-14 | 2019-09-06 | 智锐达仪器科技南通有限公司 | A kind of Detecting Pesticide pulvis and preparation method, cuvette, detection method |
| CN112326646A (en) * | 2020-11-03 | 2021-02-05 | 童超 | Pesticide residue detection method for food detection |
| CN115656531A (en) * | 2022-12-08 | 2023-01-31 | 深圳市赛特罗生物医疗技术有限公司 | Integrated pesticide residue standardized rapid detection box and detection method |
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