CN110487977A - A kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method - Google Patents
A kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method Download PDFInfo
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- C—CHEMISTRY; METALLURGY
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q1/46—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase involving cholinesterase
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- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
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- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
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- G01N33/02—Food
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- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
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Abstract
The present invention relates to Practice for Pesticide Residue in Agricultural Products detection technique fields, specifically disclose a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method, comprising the following steps: pericarp removing surface；Cleaning liquid concentration；Separate fruit carpopodium, separate fruit upper and lower ends pericarp, separation fruit among pericarp and pulp；Prepare sample to be tested stoste；Preparation of reagents, the preparation including buffer, substrate solution, chromophoric solution and enzyme solutions；The detection of control sample and sample.In detection method of the invention, to the upper, middle and lower of fruit, it is inside and outside do careful detection and divide, therefore improve the fine degree of each position Detecting Pesticide of entire fruit；Individual micronization processes are carried out to the various pieces of fruit in detection method of the invention, entire testing result can form the comparison of a system, therefore reduce the error of its detection, improve the precision of detection.
Description
Technical field
The present invention relates to Practice for Pesticide Residue in Agricultural Products detection technique field, specially a kind of Practice for Pesticide Residue in Agricultural Products is accurately detected
Method.
Background technique
The pesticide residue content of agricultural product is to detect an important indicator of agricultural product quality, the pesticide residue in agricultural product
The edible safety that consumer can be seriously affected after exceeded will cause consumer and cause a disease or even dead when serious, therefore every country
There is stringent standard formulation to the pesticide residue with agricultural product, the pesticide residue how research reduces agricultural product becomes a weight
The scientific research subject under discussion wanted, and the detection of Residual Pesticides in Farm Produce just becomes an important inspection foundation for improving agricultural production quality, it is existing
There is the detection method for Practice for Pesticide Residue in Agricultural Products in technology to have very much, is broadly divided into according to its principle: biochemical assay
With chromatographic detection, although existing detection method can determine the pesticide residue in agricultural product to a certain extent,
There is also following shortcomings in actual use for they:
It, can not be each for fruit 1. detection position is excessively single for pulp more fruit such as apple, pear, peaches
Partial pesticide residue degree does corresponding fine detection；
2. it is too simple to the processing of test object during actually detected, the accurate comparison an of system can not be formed,
Therefore testing result reduces its whole detection precision there are biggish error.
Summary of the invention
The purpose of the present invention is to provide a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting methods, to solve above-mentioned background technique
The problem of middle proposition.
To achieve the above object, the invention provides the following technical scheme: a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method,
This method specifically includes the following steps:
Fruit is placed into sonic cleaning slot by step 1, pericarp removing surface, and 20-25 DEG C of temperature is added in groove body
Water did not had fruit 1cm height to be limited, and after impregnating 3-5 minutes, using ultrasonic cleaning 2-3 minutes, then took out fruit with the water surface
Fruit face is dried, while cleaning liquid being taken out spare；
Step 2, filtering and concentrating stir 5-10 minutes after the cleaning liquid in step 1 is heated to 40-45 DEG C, then by solution
The wax layer on surface layer filters out, while the impurity in filtrate being filtered out, and leaves clear liquid, will be extra in filtrate using freeze drier
Moisture takes out, and adjusts the concentration of filtrate spare to then refrigerating filtrate within the scope of 0.01%-0.1%, refrigerated storage temperature control exists
2-4℃；
Step 3 separates the pericarp at fruit carpopodium, and the carpopodium of fruit is removed, then will be within the scope of the 1cm near carpopodium
Pericarp is cut, and pays attention to peel thickness control in 1-2mm；
Step 4 separates the pericarp at fruit areola, then the pericarp within the scope of 1cm near fruit navel is cut, and pays attention to pericarp
Thickness control is in 1-2mm；
Step 5 separates pulp and pericarp among fruit, the pericarp of remainder is cut using beam knife, pays attention to pericarp
Then pulp is pressed subcutaneous 2-5mm in 1-1.5mm by thickness control, subcutaneous 5-10mm is cut, pays attention to pulp being cut into 0.5-
The square of 1cm size；
Step 6, by Step 3: the pericarp and pulp cut in step 4 and step 5 are respectively put into individual blender,
It is respectively crushed, pericarp and pulp is ground into broken pulp, are then poured into pulp in centrifuge tube respectively, and to centrifugation
2-3 times of pulp of deionized water is added in pipe, impregnates 10-20 minutes, is put into centrifuge after 10-20 minutes, by centrifuge tube
It takes out, and respectively filters out the residue in slurries, take supernatant liquor as sample to be tested stoste；
Buffering agents are mixed with distilled water that shake up into buffer spare by step 7, preparation of reagents；It will be labeled as the bottle of substrate
The middle appropriate distilled water of addition shakes up into substrate solution, and stored refrigerated spare；Color developing agent is mixed with appropriate amount of buffer solution and is shaken up into
Chromophoric solution, and it is stored refrigerated spare；Detection enzyme matches to mix with appropriate amount of buffer solution shakes up into enzyme solutions, stored refrigerated spare；
Step 8, control sample detection, takes enzyme solutions 100ul to mix with buffer 2.5ml and shakes up, chromophoric solution is then added
100ul is mixed with substrate solution 20ul shake up after be put into cuvette machine testing, and record the absorbance of reaction 3min
Changing value size.
Sample to be tested stoste is first mixed that shake up into sample detection liquid spare with buffer, takes enzyme by step 9, sample detection
Solution 100ul is mixed with sample detection liquid 2.5ml and is shaken up and inhibit 10-15min, and chromophoric solution 100ul and substrate is then added
Upper machine testing is put into cuvette in solution 100ul mixing after shaking up, and records the absorbance change value size of reaction 3min.
Preferably, in step 6, pay attention to putting into 1/5th that pulp in centrifuge tube is centrifuge tube volume, it is used from
Heart pipe is with a scale, the sharp bottom of 10-15ml, even cap-type centrifuge tube.
Preferably, in step 6, the crushing duration of pericarp and pulp is respectively 8-10 minutes and 5-8 minutes, crushes consequence
Pulp particle degree size in slurry is controlled in 60-80 mesh.
Preferably, in step 7, selected buffering agents are by anhydrous potassium dihydrogen phosphate and potassium dihydrogen phosphate with the ratio of 15:4
It mixes again, and the specific gravity of buffering agents and distilled water is 19:125 in buffer.
Preferably, in step 7, substrate is acetylthiocholine, and the specific gravity of substrate and distilled water is 1:120, and substrate is molten
The refrigerated storage temperature of liquid is controlled at 2-4 DEG C.
Preferably, in step 7, selected color developing agent is by dithiobis-nitrobenzoic acid and sodium bicarbonate with the specific gravity of 10:1
Mixing, and the proportion specific gravity of color developing agent and buffer is 1:116, the refrigerated storage temperature of chromophoric solution is controlled at 2-4 DEG C.
Preferably, in step 7, selected detection enzyme is acetylcholinesterase, and detects enzyme and buffer according to detection enzyme
Active situation is matched, and guarantees that absorbance change value of the acetylcholinesterase in the enzyme solutions after preparing in 3min should be big
In 0.3, the refrigerated storage temperature of enzyme solutions is controlled at 2-4 DEG C.
Compared with prior art, the beneficial effects of the present invention are:
1. in detection method of the invention, to the upper, middle and lower of fruit, it is inside and outside do careful detection and divide, therefore improve
The fine degree of each position Detecting Pesticide of entire fruit；
2. carrying out individual micronization processes to the various pieces of fruit in detection method of the invention, entire testing result can be with shape
At the comparison of a system, therefore the error of its detection is reduced, improves the precision of detection.
Specific embodiment
The technical scheme in the embodiments of the invention will be clearly and completely described below, it is clear that described implementation
Example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is common
Technical staff's every other embodiment obtained without making creative work belongs to the model that the present invention protects
It encloses.
Embodiment one
The present invention provides a kind of technical solution: a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method, and agricultural product to be checked are the spray of fruit face
The apple of medicine after a week, and not rainfall in one week, pesticide residue method in this apple specifically includes the following steps:
Apple is placed into sonic cleaning slot by step 1, pericarp removing surface, and 25 DEG C of warm water is added in groove body,
So that the water surface was not had apple surface 1cm height, impregnates after five minutes, using ultrasonic cleaning 3 minutes, then take out fruit for fruit face
Dry, while cleaning liquid being taken out spare, pay attention to before cleaning cutting short the carpopodium of apple, avoid because too long carpopodium bending with
Pericarp fitting causes pericarp impaired, influences testing result, when wiping fruit, guarantees that wiping dynamics is uniform, not hurt pericarp,
Influence testing result；
Step 2, filtering and concentrating stir 8 minutes after the cleaning liquid in step 1 is heated to 42 DEG C, in the process apple table
The wax layer of layer can melt and swim in solution surface layer, then filter out the wax layer on solution surface layer, in order to avoid the silt in filtrate
Measurement result is influenced, while the impurity in filtrate being filtered out, clear liquid is left, using freeze drier, by water extra in filtrate
Divide and take out, paying attention to, which controls the overall volume of filtrate, adjusts the concentration of filtrate to 0.01%-0.1% range
It is interior, then filtrate is refrigerated it is spare, refrigerated storage temperature control at 4 DEG C；
Step 3 separates the pericarp at apple carpopodium, and the carpopodium of apple is removed, because the carpopodium of apple is non-edible part,
Therefore not as the necessary detection part of pesticide residue, and apple carpopodium is in when spraying insecticide, and pesticide is easy to be detained here
In the pit of formation, therefore pesticide residue easy to form and non-easy cleaning, therefore pesticide residue level herein need to be with apple
The pesticide residue level at intermediate position will be different, and need to carry out difference detection, then will be within the scope of the 1cm near carpopodium
Pericarp is cut, and in cutting process remittance, is paid attention to cutting steadily, is guaranteed the continuity of pericarp, the peel thickness cut is 1.5mm；
Step 4 separates the pericarp at apple areola, and during spraying insecticide, medical fluid can be slipped to apple along apple surface
The areola bottom of apple bottom can form spot, therefore pesticide residue journey herein in the bottom of areola after usual medical fluid volatilization
Spending will be different with the pesticide residue degree at the intermediate position of apple, need to carry out difference detection, then will be near apple areola
Pericarp within the scope of 1cm is cut, and is paid attention to the stationarity for keeping fortune knife in cutting process, is guaranteed the coherent of the pericarp under cutting as far as possible
Property, the peel thickness cut is 1.6mm；
Step 5 separates pulp and pericarp among apple, the pericarp of remainder is cut using beam knife, in the mistake peeled
Cheng Zhong pays attention to the consistency for guaranteeing the peel thickness under cutting, and the peel thickness cut is 1.3mm, then by pulp by subcutaneous
3mm, subcutaneous 6mm are cut, and pay attention to the square that pulp is cut into 0.6cm size；
Step 6, by Step 3: the pericarp and pulp cut in step 4 and step 5 are respectively put into individual blender,
It is respectively crushed, the crushing duration of pericarp and pulp is respectively 10 minutes and 8 minutes, and pericarp and pulp are ground into broken fruit
Slurry, the pulp particle degree size after crushing in pulp control in 60-80 mesh, are then poured into pulp in centrifuge tube respectively, and
3 times of pulp of deionized water is added into centrifuge tube, impregnates 15 minutes, is put into centrifuge after 15 minutes, centrifuge tube is taken
Out, and respectively the residue in slurries is filtered out, takes supernatant liquor as sample to be tested stoste, pay attention to putting into the fruit in centrifuge tube
Slurry is 1/5th of centrifuge tube volume, and centrifuge tube used is with a scale, the sharp bottom of 10ml, even cap-type centrifuge tube；
Step 7, preparation of reagents take anhydrous potassium dihydrogen phosphate 15g and potassium dihydrogen phosphate to be mixed and made into buffer examination with 4g respectively
The buffering agents of preparation, are then mixed that shake up into buffer spare by agent with 1250ml distilled water；Select 30mg thioacetyl
Then the distilled water for being labeled as adding 3.6ml in the bottle of substrate is shaken up into substrate solution as substrate by choline, and at 4 DEG C
At a temperature of it is stored refrigerated spare；Take dithiobis-nitrobenzoic acid 200mg and sodium bicarbonate 19.5mg that color developing agent is made respectively, it will
Color developing agent mixes with 25ml buffer and shakes up into chromophoric solution, and 4 DEG C at a temperature of it is stored refrigerated spare；Select acetylcholine
Esterase matches to mix as detection enzyme with suitable buffer shakes up into enzyme solutions, guarantees the acetylcholine in the enzyme solutions after preparing
Absorbance change value of the esterase in 3min should be greater than 0.3, then 4 DEG C at a temperature of it is stored refrigerated spare；
Step 8, control sample detection, takes enzyme solutions 100ul to mix with buffer 2.5ml and shakes up, chromophoric solution is then added
100ul is mixed with substrate solution 20ul shake up after be put into cuvette machine testing, and record the absorbance of reaction 3min
Changing value size.
Sample to be tested stoste is first mixed that shake up into sample detection liquid spare by step 9, sample detection with buffer respectively,
Pay attention to not obscuring sample detection liquid prepared by apple various pieces when preparing sample detection liquid, take enzyme solutions 100ul with
Sample detection liquid 2.5ml mixing shakes up and inhibits 15min, and then addition chromophoric solution 100ul is mixed with substrate solution 100ul shakes
It is put into cuvette upper machine testing after even, and records the absorbance change value size of reaction 3min, successively by each portion of apple
The sample detection liquid of position is detected by above-mentioned steps, and be will test result and uniformly recorded.
Embodiment two
The present invention provides a kind of technical solution: a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method, and agricultural product to be checked are the spray of fruit face
The pear of medicine after a week, and not rainfall in one week, pesticide residue method in this pear specifically includes the following steps:
Pear is placed into sonic cleaning slot by step 1, pericarp removing surface, and 22 DEG C of warm water is added in groove body,
So that the water surface was not had pear surface 1cm height, after impregnating 4 minutes, using ultrasonic cleaning 2 minutes, then takes out fruit for fruit face
Dry, while cleaning liquid being taken out spare, pay attention to before cleaning cutting short the carpopodium of pear, avoid because too long carpopodium bending with
Pericarp fitting causes pericarp impaired, influences testing result, when wiping fruit, guarantees that wiping dynamics is uniform, not hurt pericarp,
Influence testing result；
Step 2, filtering and concentrating stir 5 minutes after the cleaning liquid in step 1 is heated to 40 DEG C, in the process pear table
The wax layer of layer can melt and swim in solution surface layer, then filter out the wax layer on solution surface layer, in order to avoid the silt in filtrate
Measurement result is influenced, while the impurity in filtrate being filtered out, clear liquid is left, using freeze drier, by water extra in filtrate
Divide and take out, paying attention to, which controls the overall volume of filtrate, adjusts the concentration of filtrate to 0.01%-0.1% range
It is interior, then filtrate is refrigerated it is spare, refrigerated storage temperature control at 4 DEG C；
Step 3 separates the pericarp at pear carpopodium, and the carpopodium of pear is removed, because the carpopodium of pear is non-edible part,
Therefore not as the necessary detection part of pesticide residue, and pear carpopodium is in when spraying insecticide, and pesticide is easy to be detained here
In the pit of formation, therefore pesticide residue easy to form and non-easy cleaning, therefore pesticide residue level herein need to be with pear
The pesticide residue level at intermediate position will be different, and need to carry out difference detection, then will be within the scope of the 1cm near carpopodium
Pericarp is cut, and in cutting process remittance, is paid attention to cutting steadily, is guaranteed the continuity of pericarp, the peel thickness cut is 1.8mm；
Step 4 separates the pericarp at pear areola, and during spraying insecticide, medical fluid can be slipped to pear along pear surface
The areola bottom of pear bottom can form spot, therefore pesticide residue journey herein in the bottom of areola after usual medical fluid volatilization
Spending will be different with the pesticide residue degree at the intermediate position of pear, need to carry out difference detection, then will be near pear areola
Pericarp within the scope of 1cm is cut, and is paid attention to the stationarity for keeping fortune knife in cutting process, is guaranteed the coherent of the pericarp under cutting as far as possible
Property, the peel thickness cut is 1.7mm；
Step 5 separates pulp and pericarp among pear, the pericarp of remainder is cut using beam knife, in the mistake peeled
Cheng Zhong pays attention to the consistency for guaranteeing the peel thickness under cutting, and the peel thickness cut is 1.5mm, then by pulp by subcutaneous
4mm, subcutaneous 8mm are cut, and pay attention to the square that pulp is cut into 0.8cm size；
Step 6, by Step 3: the pericarp and pulp cut in step 4 and step 5 are respectively put into individual blender,
It is respectively crushed, the crushing duration of pericarp and pulp is respectively 8 minutes and 5 minutes, and pericarp and pulp are ground into broken fruit
Slurry, the pulp particle degree size after crushing in pulp control in 60-80 mesh, are then poured into pulp in centrifuge tube respectively, and
2 times of pulp of deionized water is added into centrifuge tube, impregnates 10 minutes, is put into centrifuge after ten minutes, centrifuge tube is taken
Out, and respectively the residue in slurries is filtered out, takes supernatant liquor as sample to be tested stoste, pay attention to putting into the fruit in centrifuge tube
Slurry is 1/5th of centrifuge tube volume, and centrifuge tube used is with a scale, the sharp bottom of 15ml, even cap-type centrifuge tube；
Step 7, preparation of reagents take anhydrous potassium dihydrogen phosphate 20g and potassium dihydrogen phosphate to be mixed and made into buffer with 5.33g respectively
The buffering agents of preparation, are then mixed that shake up into buffer spare by reagent with 1666.67ml distilled water；Select 40mg thio
Then the distilled water for being labeled as adding 4.8ml in the bottle of substrate is shaken up into substrate solution as substrate by acetylcholine, and 4
It is stored refrigerated spare at a temperature of DEG C；Take dithiobis-nitrobenzoic acid 266.67mg and sodium bicarbonate 26mg that colour developing is made respectively
Color developing agent is mixed with 33.33ml buffer and shakes up into chromophoric solution by agent, and 4 DEG C at a temperature of it is stored refrigerated spare；It selects
Acetylcholinesterase matches to mix as detection enzyme with suitable buffer shakes up into enzyme solutions, guarantees in the enzyme solutions after preparing
Absorbance change value of the acetylcholinesterase in 3min should be greater than 0.3, then 4 DEG C at a temperature of it is stored refrigerated spare；
Step 8, control sample detection, takes enzyme solutions 100ul to mix with buffer 2.5ml and shakes up, chromophoric solution is then added
100ul is mixed with substrate solution 20ul shake up after be put into cuvette machine testing, and record the absorbance of reaction 3min
Changing value size.
Sample to be tested stoste is first mixed that shake up into sample detection liquid spare by step 9, sample detection with buffer respectively,
Pay attention to not obscuring sample detection liquid prepared by pear various pieces when preparing sample detection liquid, take enzyme solutions 100ul with
Sample detection liquid 2.5ml mixing shakes up and inhibits 15min, and then addition chromophoric solution 100ul is mixed with substrate solution 100ul shakes
It is put into cuvette upper machine testing after even, and records the absorbance change value size of reaction 3min, successively by each portion of pear
The sample detection liquid of position is detected by above-mentioned steps, and be will test result and uniformly recorded.
Embodiment three
The present invention provides a kind of technical solution: a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method, and agricultural product to be checked are the spray of fruit face
The peach of medicine after a week, and not rainfall in one week, pesticide residue method in this peach specifically includes the following steps:
Peach is placed into sonic cleaning slot by step 1, pericarp removing surface, and 23 DEG C of warm water is added in groove body,
So that the water surface was not had peach surface 1cm height, after impregnating 6 minutes, using ultrasonic cleaning 4 minutes, then takes out fruit for fruit face
Dry, while cleaning liquid being taken out spare, pay attention to before cleaning cutting short the carpopodium of peach, avoid because too long carpopodium bending with
Pericarp fitting causes pericarp impaired, influences testing result, when wiping fruit, guarantees that wiping dynamics is uniform, not hurt pericarp,
Influence testing result；
Step 2, filtering and concentrating stir 10 minutes after the cleaning liquid in step 1 is heated to 45 DEG C, in the process peach table
The wax layer of layer can melt and swim in solution surface layer, then filter out the wax layer on solution surface layer, in order to avoid the silt in filtrate
Measurement result is influenced, while the impurity in filtrate being filtered out, clear liquid is left, using freeze drier, by water extra in filtrate
Divide and take out, paying attention to, which controls the overall volume of filtrate, adjusts the concentration of filtrate to 0.01%-0.1% range
It is interior, then filtrate is refrigerated it is spare, refrigerated storage temperature control at 4 DEG C；
Step 3 separates the pericarp at peach carpopodium, and the carpopodium of peach is removed, because the carpopodium of peach is non-edible part,
Therefore not as the necessary detection part of pesticide residue, and peach carpopodium is in when spraying insecticide, and pesticide is easy to be detained here
In the pit of formation, therefore pesticide residue easy to form and non-easy cleaning, therefore pesticide residue level herein need to be with peach
The pesticide residue level at intermediate position will be different, and need to carry out difference detection, then will be within the scope of the 1cm near carpopodium
Pericarp is cut, and in cutting process remittance, is paid attention to cutting steadily, is guaranteed the continuity of pericarp, the peel thickness cut is 2mm；
Step 4 separates the pericarp of peach peach tip, and during spraying insecticide, medical fluid can be slipped to peach along peach surface
The peach tip bottom of peach bottom can form spot, therefore pesticide residue journey herein in the bottom of peach tip after usual medical fluid volatilization
Spending will be different with the pesticide residue degree at the intermediate position of peach, need to carry out difference detection, then will be near peach peach tip
Pericarp within the scope of 1cm is cut, and is paid attention to the stationarity for keeping fortune knife in cutting process, is guaranteed the coherent of the pericarp under cutting as far as possible
Property, the peel thickness cut is 2mm；
Step 5 separates pulp and pericarp among peach, the pericarp of remainder is cut using beam knife, in the mistake peeled
Cheng Zhong pays attention to the consistency for guaranteeing the peel thickness under cutting, and the peel thickness cut is 1.5mm, then by pulp by subcutaneous
4mm, subcutaneous 8mm are cut, and pay attention to the square that pulp is cut into 1cm size；
Step 6, by Step 3: the pericarp and pulp cut in step 4 and step 5 are respectively put into individual blender,
It is respectively crushed, the crushing duration of pericarp and pulp is respectively 8 minutes and 5 minutes, and pericarp and pulp are ground into broken fruit
Slurry, the pulp particle degree size after crushing in pulp control in 60-80 mesh, are then poured into pulp in centrifuge tube respectively, and
2-3 times of pulp of deionized water is added into centrifuge tube, impregnates 12 minutes, is put into centrifuge after 15 minutes, by centrifuge tube
It takes out, and respectively filters out the residue in slurries, take supernatant liquor as sample to be tested stoste, pay attention to putting into centrifuge tube
Pulp is 1/5th of centrifuge tube volume, and centrifuge tube used is with a scale, the sharp bottom of 15ml, even cap-type centrifuge tube；
Step 7, preparation of reagents take anhydrous potassium dihydrogen phosphate 12g and potassium dihydrogen phosphate to be mixed and made into buffer examination with 3.2g respectively
The buffering agents of preparation, are then mixed that shake up into buffer spare by agent with 1000ml distilled water；Select 24mg thioacetyl
Then the distilled water for being labeled as adding 2.88ml in the bottle of substrate is shaken up into substrate solution as substrate by choline, and at 4 DEG C
At a temperature of it is stored refrigerated spare；Take dithiobis-nitrobenzoic acid 160mg and sodium bicarbonate 15.6mg that color developing agent is made respectively, it will
Color developing agent mixes with 20ml buffer and shakes up into chromophoric solution, and 4 DEG C at a temperature of it is stored refrigerated spare；Select acetylcholine
Esterase matches to mix as detection enzyme with suitable buffer shakes up into enzyme solutions, guarantees the acetylcholine in the enzyme solutions after preparing
Absorbance change value of the esterase in 3min should be greater than 0.3, then 4 DEG C at a temperature of it is stored refrigerated spare；
Step 8, control sample detection, takes enzyme solutions 100ul to mix with buffer 2.5ml and shakes up, chromophoric solution is then added
100ul is mixed with substrate solution 20ul shake up after be put into cuvette machine testing, and record the absorbance of reaction 3min
Changing value size.
Sample to be tested stoste is first mixed that shake up into sample detection liquid spare by step 9, sample detection with buffer respectively,
Pay attention to not obscuring sample detection liquid prepared by peach various pieces when preparing sample detection liquid, take enzyme solutions 100ul with
Sample detection liquid 2.5ml mixing shakes up and inhibits 10-15min, and it is mixed that chromophoric solution 100ul and substrate solution 100ul is then added
Upper machine testing is put into cuvette in conjunction after shaking up, and records the absorbance change value size of reaction 3min, successively by peach
The sample detection liquid at each position is detected by above-mentioned steps, and be will test result and uniformly recorded:
As seen from the above table, the Practice for Pesticide Residue in Agricultural Products result of the measurement of above-mentioned three groups of embodiments is than comparative example that commonsense method measures
The measurement direction of middle Practice for Pesticide Residue in Agricultural Products result wants more more, and the persticide residue at each position of fruit measured has
One detailed statistical result provides good data support to improve the pesticide residue content of fruit.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (7)
1. a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method, which is characterized in that this method specifically includes the following steps:
Fruit is placed into sonic cleaning slot by step 1, pericarp removing surface, and 20-25 DEG C of temperature is added in groove body
Water did not had fruit 1cm height to be limited, and after impregnating 3-5 minutes, using ultrasonic cleaning 2-3 minutes, then took out fruit with the water surface
Fruit face is dried, while cleaning liquid being taken out spare；
Step 2, filtering and concentrating stir 5-10 minutes after the cleaning liquid in step 1 is heated to 40-45 DEG C, then by solution
The wax layer on surface layer filters out, while the impurity in filtrate being filtered out, and leaves clear liquid, will be extra in filtrate using freeze drier
Moisture takes out, and adjusts the concentration of filtrate spare to then refrigerating filtrate within the scope of 0.01%-0.1%, refrigerated storage temperature control exists
2-4℃；
Step 3 separates the pericarp at fruit carpopodium, and the carpopodium of fruit is removed, then will be within the scope of the 1cm near carpopodium
Pericarp is cut, and pays attention to peel thickness control in 1-2mm；
Step 4 separates the pericarp at fruit areola, then the pericarp within the scope of 1cm near fruit navel is cut, and pays attention to pericarp
Thickness control is in 1-2mm；
Step 5 separates pulp and pericarp among fruit, the pericarp of remainder is cut using beam knife, pays attention to pericarp
Then pulp is pressed subcutaneous 2-5mm in 1-1.5mm by thickness control, subcutaneous 5-10mm is cut, pays attention to pulp being cut into 0.5-
The square of 1cm size；
Step 6, by Step 3: the pericarp and pulp cut in step 4 and step 5 are respectively put into individual blender,
It is respectively crushed, pericarp and pulp is ground into broken pulp, are then poured into pulp in centrifuge tube respectively, and to centrifugation
2-3 times of pulp of deionized water is added in pipe, impregnates 10-20 minutes, is put into centrifuge after 10-20 minutes, by centrifuge tube
It takes out, and respectively filters out the residue in slurries, take supernatant liquor as sample to be tested stoste；
Buffering agents are mixed with distilled water that shake up into buffer spare by step 7, preparation of reagents；It will be labeled as the bottle of substrate
The middle appropriate distilled water of addition shakes up into substrate solution, and stored refrigerated spare；Color developing agent is mixed with appropriate amount of buffer solution and is shaken up into
Chromophoric solution, and it is stored refrigerated spare；Detection enzyme matches to mix with appropriate amount of buffer solution shakes up into enzyme solutions, stored refrigerated spare；
Step 8, control sample detection, takes enzyme solutions 100ul to mix with buffer 2.5ml and shakes up, chromophoric solution is then added
100ul is mixed with substrate solution 20ul shake up after be put into cuvette machine testing, and record the absorbance of reaction 3min
Changing value size；
Sample to be tested stoste is first mixed that shake up into sample detection liquid spare with buffer, takes enzyme solutions by step 9, sample detection
100ul is mixed with sample detection liquid 2.5ml and is shaken up and inhibit 10-15min, and chromophoric solution 100ul and substrate solution is then added
Upper machine testing is put into cuvette in 100ul mixing after shaking up, and records the absorbance change value size of reaction 3min.
2. a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method according to claim 1, it is characterised in that: in step 6,
Paying attention to putting into the pulp in centrifuge tube is 1/5th of centrifuge tube volume, and centrifuge tube used is the with a scale, sharp of 10-15ml
Bottom, even cap-type centrifuge tube.
3. a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method according to claim 1, it is characterised in that: in step 6,
The crushing duration of pericarp and pulp is respectively 8-10 minutes and 5-8 minutes, the pulp particle degree size control after crushing in pulp
In 60-80 mesh.
4. a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method according to claim 1, it is characterised in that: in step 7,
Selected buffering agents are mixed by anhydrous potassium dihydrogen phosphate with potassium dihydrogen phosphate with the specific gravity of 15:4, and buffer tries in buffer
The specific gravity of agent and distilled water is 19:125.
5. a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method according to claim 1, it is characterised in that: in step 7,
Substrate is acetylthiocholine, and the specific gravity of substrate and distilled water is 1:120, and the refrigerated storage temperature of substrate solution is controlled at 2-4 DEG C.
6. a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method according to claim 1, it is characterised in that: in step 7,
Selected color developing agent is mixed by dithiobis-nitrobenzoic acid and sodium bicarbonate with the specific gravity of 10:1, and color developing agent and buffer are matched
It is 1:116 than specific gravity, the refrigerated storage temperature of chromophoric solution is controlled at 2-4 DEG C.
7. a kind of Practice for Pesticide Residue in Agricultural Products accurate detecting method according to claim 1, it is characterised in that: in step 7,
Selected detection enzyme is acetylcholinesterase, and detects enzyme and buffer and matched according to the active situation of detection enzyme, guarantees to match
Absorbance change value of the acetylcholinesterase in enzyme solutions in 3min after system should be greater than 0.3, the refrigerated storage temperature of enzyme solutions
Control is at 2-4 DEG C.
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| CN115855888A (en) * | 2023-02-14 | 2023-03-28 | 广东省农业科学院植物保护研究所 | Litchi residual pesticide detection system and evaluation method |
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