CN108020609A - Quick determination method that is a kind of while measuring 34 kinds of pesticide residues in fruits and vegetables - Google Patents
Quick determination method that is a kind of while measuring 34 kinds of pesticide residues in fruits and vegetables Download PDFInfo
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Abstract
The present invention relates to field of pesticide detection.Purpose is to provide a kind of LC MS detection methods that can measure 34 kinds of garden stuff pesticide residues at the same time, to realize the method that can analyze 34 kinds of pesticide residues at the same time without multiple detection methods, meets the requirement of detection work rapidly and efficiently；A kind of method of garden stuff pesticide residue detection sample pre-treatments is additionally provided, this method has the characteristics that the used time is few, extraction effect is good.Technical solution is：The extractant of sample pre-treatments during a kind of detection for garden stuff pesticide residue, includes the raw material of following parts by weight：08 parts of acetonitrile, 05 parts of acetone, 03 parts of 1 butyl, 3 methylimidazole dicyan amine salt；Quick determination method that is a kind of while measuring 34 kinds of pesticide residues in fruits and vegetables, follows the steps below：1) standard curve is made；2) sample to be tested is measured.
Description
Technical field
The present invention relates to field of pesticide detection, more particularly to a kind of LC-MS measures 34 kinds of pesticide residues in fruits and vegetables at the same time
Method.
Background technology
With the development of agriculture industrialization, the production of agricultural product is increasingly dependent on the external sources such as pesticide, antibiotic and hormone
Material.Pesticide residue is exceeded, influences consumer's edible safety, consumer can be caused to cause a disease when serious, develop it is abnormal, even
Directly result in and be poisoned to death.In order to ensure food safety, the pesticide residue in keeping under strict supervision close inspection food, maximally efficient method it
First, strengthen the dynamics to Pesticide Residues detection.
Carbofuran in vegetable and fruit, Methomyl, carbaryl, Aldicarb, carbendazim, imidacloprid, oxamyl, Acetamiprid, pyrrole
Aphid ketone, chloropyuril, Propamocarb, demeton, Spanon, Boscalid, tebufenozide, cyazofamid, imidaclothiz, diafenthiuron, thiophene mite
Ketone, fenazaquin, thiacloprid, Chem hoe, thiophanate-methyl, probenazole, zoxamide, imazalil, Tebuconazole, diethofencarb, fluorine pyrrole
The spirit of first standing grain, Diacloden, paclobutrazol, diazinon, triazolone and indoxacarb are common pesticide residue, are current pesticide residues
The Common item required in detection.But existing coherent detection standard is only capable of detecting several therein, it is impossible to is carried out at the same time point
Analysis；As GB/T 20770-2008 be only capable of detection 34 kinds the inside 20 kinds, GB/T 23584-2009 be only capable of detection 34 kinds the inside
1 kind, NY/T 1453-2007 are only capable of 7 kinds of 34 kinds of the insides of detection, and NY/T 1720-2009 are only capable of 2 kinds of 34 kinds of the insides of detection,
GB 23200.34-2016 are only capable of 15 kinds of 34 kinds of the insides of detection, and SN/T 2560-2010 are only capable of 4 kinds of 34 kinds of the insides of detection,
GB/T 20769-2008 are only capable of 18 kinds etc. of 34 kinds of the insides of detection.Therefore in detection process, if necessary to inspection Bi Suoyou
Mesh, then need to use different methods, and work efficiency is very low, and the pesticide residue being unfavorable in quickly detection fruits and vegetables, have impact on
Circulation of the fruits and vegetables in market.
The content of the invention
The defects of the purpose of the present invention is overcoming above-mentioned technology, there is provided a kind of to measure 34 kinds of garden stuff pesticide residues at the same time
LC-MS detection methods, to realize the method that can analyze 34 kinds of pesticide residues at the same time without multiple detection methods, meet inspection
Survey the requirement of work rapidly and efficiently.
The present invention also aims to provide a kind of method of garden stuff pesticide residue detection sample pre-treatments, this method has
The characteristics of used time is few, extraction effect is good.
Technical solution provided by the invention is：
The extractant of sample pre-treatments during a kind of detection for garden stuff pesticide residue, includes the raw material of following parts by weight：0-8
The acetonitrile, 0-5 parts of acetone, 0-3 parts of 1- butyl -3- methylimidazole dicyan amine salt of part.
Preferably, the mass ratio in the extractant between acetonitrile, acetone and 1- butyl -3- methylimidazole dicyan amine salt is
2-8：1-5： 0-3.
A kind of sample-pretreating method detected using foregoing extractant to garden stuff pesticide residue, comprises the following steps：
1) fruits and vegetables homogenised sample to be measured (without dilution or emulsion) is mixed with foregoing extractant, obtains the first mixing
Thing；
2) the first mixture obtained in step 1) is mixed with sodium chloride and anhydrous magnesium sulfate, obtains the second mixture；
3) centrifugal treating is carried out to the second mixture, takes supernatant；
4) take the supernatant in step 3), carry out nitrogen and be blown to dry, then add methanol solution and redissolve, obtain the 3rd mixing
Thing；
5) the 3rd mixture in step 4) is mixed with PSA, C18, Florey diatomite and magnesium sulfate, obtains
Four mixtures；
6) lysate of the 4th mixture in step 5) is taken, filtering with microporous membrane is carried out, obtains the sample for detection.
The quality of fruits and vegetables homogenised sample to be measured in the step 1) is 0.1-10g, preferably 0.1-5g；
The quality of extractant in the step 1) is 0.1-20mL, preferably 0.1-10mL；
The mass ratio of sodium chloride and anhydrous magnesium sulfate in the step 2) is 1：1-5；The matter of sodium chloride, anhydrous magnesium sulfate
The sum of amount, the mass ratio with first mixture are 1：1-10；
Preferably, the mass ratio of the sodium chloride in the step 2) and anhydrous magnesium sulfate is 1：3-5；
The mass fraction of methanol solution is 0.1-80%, preferably 30-60% in the step 4)；Selected methanol solution adds
Dosage is 0.1-5mL；
The quality sum of PSA, C18, Florey diatomite and magnesium sulfate in the step 5), with the 3rd mixture
Mass ratio is 1：5-20；Mass ratio between the PSA, C18, Florey diatomite and magnesium sulfate is 1：0-5：0-5：0-5,
It is preferred that 1：0.5-2：0.5-2：0.5-2：0.5-2；
The aperture of miillpore filter in the step 6) is 0.22-0.45 μm.
Quick determination method that is a kind of while measuring 34 kinds of pesticide residues in fruits and vegetables, follows the steps below：
1) standard curve is made
A variety of agriculture residual standard product are respectively placed in volumetric flask, add acetonitrile dissolving and constant volume, are configured to single mark storing solution；Take
Single mark storing solution of the residual compound of each agriculture of certain volume is mixed, add methanol aqueous solution and be diluted to middle interstitial fluid, then is taken
Interstitial fluid is diluted the hybrid standard working solution for being configured to a variety of concentration, injects on HPLC-MS/MS and measures, obtains Multiple Pesticides
The standard curve of residual compounds；
2) sample to be tested is measured
Pre-treatment first is carried out to sample to be tested, then upper machine measure obtains the total ion chromatogram of sample to be tested；Then will
Obtained sample to be tested inspection data is detected, the remaining concentration of pesticide in sample to be tested is calculated by standard curve.
The method for carrying out pre-treatment in the step 2) to sample to be tested is as follows：
1) fruits and vegetables sample to be measured is mixed with foregoing extractant；
2) mixture obtained in step 1) is mixed with sodium chloride and anhydrous magnesium sulfate；
3) centrifugal treating is carried out to the mixture obtained in step 2), takes supernatant；
4) take the supernatant liquid nitrogen in step 3) to be blown to dry, then add methanol solution and redissolve；
5) mixture in step 4) is mixed with PSA, C18, Florey diatomite and magnesium sulfate；
6) take the lysate of the mixture in step 5) to carry out filtering with microporous membrane, obtain the sample for detection.
The beneficial effects of the invention are as follows：
Extractant provided by the invention is ionic liquid complex solvent, can be effective for the pre-treatment of garden stuff pesticide residue
The extraction of garden stuff pesticide residue is improved, and reduces the usage amount of organic extraction solvent.Experiment shows, is extracted using this method
Agent is used only 10mL and can reach testing requirements, about reduces 75%, can effectively reduce the generation of organic liquid waste, be conducive to protect
Retaining ring border；
Extractant provided by the invention is used for the sample pre-treatments of garden stuff pesticide residue, can extract at the same time in 34 kinds of fruits and vegetables
Pesticide residue, can avoid carrying out the pesticide residue analysis of same fruits and vegetables using different examination criterias.
The pre-treating method provided by the invention that garden stuff pesticide residue is carried out using said extracted agent, it is only necessary to 90 minutes
At the same time complete 34 kinds of garden stuff pesticide residues detection work, with existing pre-treating method QuECHERS (Quick, Easy,
Cheap, Effective, Rugged, Safe) used in the time compare, reduce 50% or so, highly shortened work
Time, improves the work efficiency of detection.
Brief description of the drawings
Fig. 1 is the total ion chromatogram that 34 kinds of pesticide residue concentration are 25.00ng/mL standard solution.
Fig. 2 is 34 kinds of pesticide residue total ion chromatograms in apple sample.
Fig. 3 is 34 kinds of pesticide residue total ion chromatograms of apple sample that additive amount is 5.0 μ g/kg standard vestigials.
Fig. 4 is 34 kinds of pesticide residue total ion chromatograms in cucumber sample.
Fig. 5 is 34 kinds of pesticide residue total ion chromatograms of cucumber sample that additive amount is 5.0 μ g/kg standard vestigials.
Fig. 6 is 34 kinds of pesticide residue total ion chromatograms in strawberry sample.
Fig. 7 is 34 kinds of pesticide residue total ion chromatograms of strawberry sample that additive amount is 5.0 μ g/kg standard vestigials.
Fig. 8 is 34 kinds of pesticide residue total ion chromatograms in tomato sample.
Fig. 9 is 34 kinds of pesticide residue total ion chromatograms of tomato sample that additive amount is 5.0 μ g/kg standard vestigials.
Figure 10 is the selection chromatography of ions figure of diafenthiuron.
Figure 11 is the selection chromatography of ions figure of Boscalid.
Figure 12 is the standard of physical curve map of diafenthiuron.
Figure 13 is the standard of physical curve map of Boscalid.
Embodiment
The present invention is further described with reference to following instance, but does not limit the present invention.
Embodiment 1
The foundation of 34 kinds of pesticide residue reference substance chromatograms
Comprise the following steps：
(1) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
A chromatographic conditions
Chromatographic column：Using Agilent SB-C18Column (2.1 × 100mm, 1.8 μm, USA)
Column temperature：35℃；
Sampling volume：5μL；Flow velocity：0.3mL/min；
Mobile phase A：0.1% formic acid water；Mobile phase B：Acetonitrile=90~10:10~90 (volume ratios), gradient elution 0~
3min, A become 50% by 90；3~5min, A are changed into 10% from 50；It is 10% that A is maintained during 5~8min；8~10min, A are by 10
It is changed into 90%.
B Mass Spectrometry Conditions
Ion gun：Electric spray ion source (ESI)；Scan mode：Cation；
Detection mode：Multiple-reaction monitoring (MRM) mode gathers；
Detector：The triple level Four bar mass spectrographs of Agilent 6460 are furnished with ESI ion guns；Wherein qualitative ion pair, quantitative
The parameters such as ion pair, collision energy refer to table 1.
The Mass Spectrometry Conditions of 34 kinds of pesticides and related chemicals in 1 fruits and vegetables of table
(2) standard curve is drawn
1) using 50% methanol aqueous solution as solvent, every kind of pesticide configuration concentration be respectively 5.00ng/mL, 10.00ng/mL,
Six kinds of standard working solutions of 25.00ng/mL, 50.00ng/mL, 75.00ng/mL and 100.0ng/mL；Then it is same by 34 kinds
The working solution mixing of sample concentration, obtains six parts of hybrid standard working solutions, by the concentration mixing mark that sampling volume is 5 μ L from low to high
Quasi- working solution carries out HPLC-MS/MS detections successively, obtains selection chromatography of ions figure (such as the fourth shown in Figure 10 of 34 parts of pesticides
Ether urea selects chromatography of ions figure, the Boscalid selection chromatography of ions figure shown in Figure 11；To save space, the choosing of remaining pesticide
Select chromatography of ions figure to be omitted)；
2) every kind of pesticide is to select ion chromatography peak area, using concentration as abscissa, to be drawn as ordinate by instrument software
The standard of physical curve of pesticide；Such as the diafenthiuron standard of physical curve map shown in Figure 12, the Boscalid material shown in Figure 13
Canonical plotting.To save space, the standard of physical curve map of remaining pesticide is omitted)；After being calculated by instrument software, obtain
Obtain the linear equation of standard of physical curve.And obtain linearly dependent coefficient R2It is worth (the linear correlation of 34 kinds of materials as shown in Table 2
0.999) coefficient is all higher than.
The linear equation and linear coefficient of 34 kinds of pesticides and related chemicals in 2 fruits and vegetables of table
The foundation of (3) 34 kinds of pesticide residue reference substance chromatograms
(pesticide, the concentration of every kind of pesticide are 25.00 to the mixed standard solution that configuration concentration is 25.00ng/mL in containing 34
Ng/mL), sampling volume carries out HPLC-MS/MS detections for 5 μ L, according to ultra performance liquid chromatography-mass spectrum/matter in step (1)
Spectral condition is measured, and obtains the total ion chromatogram of 34 kinds of pesticide residue reference substances, as shown in Figure 1.
The measure of 34 kinds of pesticide residue contents in 2 apple sample of embodiment
Comprise the following steps：
(1) test sample is prepared
It is accurate to weigh the apple sample 5.00g for smashing processing, 10mL extractants are added, are separately added into after homogeneous homogenate 1min
2 g sodium chloride and anhydrous magnesium sulfate, continue to centrifuge 5min under the conditions of 4000r/min after being homogenized 2min.Take supernatant 5mL extremely
Than in straight tube, nitrogen is blown to dry under 40 DEG C of water-baths, adds 2mL50% methanol aqueous solutions and redissolve, be vortexed after mixing and add 50mgPSA
Powder, 50mg anhydrous magnesium sulfates, 50mg florisil silicas and 50mgC18Powder, is vortexed again for crossing 0.22 μm of filter membrane after mixing,
Obtain test sample.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
A chromatographic conditions
Chromatographic column：Using Agilent SB-C18Column (2.1 × 100mm, 1.8 μm, USA)
Column temperature：35℃；
Sampling volume：5μL；Flow velocity：0.3mL/min；
Mobile phase A：0.1% formic acid water；Mobile phase B：Acetonitrile=90~10:10~90 (volume ratios), gradient elution 0~
3min, A become 50% by 90；3~5min, A are changed into 10% from 50；It is 10% that A is maintained during 5~8min；8~10min, A are by 10
It is changed into 90%..
B Mass Spectrometry Conditions
Ion gun：Electric spray ion source (ESI)；Scan mode：Cation；
Detection mode：Multiple-reaction monitoring (MRM) mode gathers；
Detector：The triple level Four bar mass spectrographs of Agilent 6460 are furnished with ESI ion guns；Wherein qualitative ion pair, quantitative
The parameters such as ion pair, collision energy refer to table 1.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain total ion chromatogram (such as Fig. 2 institutes
Show).It is compared by instrument software comparison chart 1, Fig. 2, shows to be not detected by 34 kinds of pesticide residues in apple test sample,
It is remaining containing measurement for pesticide in the apple sample of 34 kinds of pesticide residues of 5.0 μ g/kg that embodiment 3 adds concentration
It is fixed
Comprise the following steps：
(1) test sample is prepared
Accurate to weigh through smashing the negative apple sample 5.00g of processing in example 2, addition concentration is in the sample
The hybrid working solution 1.0mL of 100.0 ng/mL, then the concentration of 34 kinds of pesticide residues is 5.0 μ g/kg in the apple sample,
Subsequent treatment obtains test sample with embodiment 2.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
With embodiment 2.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain total ion chromatogram (such as Fig. 3 institutes
Show).Sample quality according to representated by test sample, it is residual to calculate 34 kinds of pesticides in test sample from the standard curve of embodiment 1
Stay the content of thing；34 kinds of object contents in apple sample are calculated in the range of 4.0~5.2 μ g/kg, the rate of recovery is
Within 80%~120%；Prove that the testing result of embodiment 2 is accurate.
The measure of 34 kinds of pesticide residue contents in 4 cucumber sample of embodiment
Comprise the following steps：
(1) test sample is prepared
It is accurate to weigh the cucumber sample 5.00g for smashing processing, 10mL extractants are added, are separately added into after homogeneous homogenate 1min
2 g sodium chloride and anhydrous magnesium sulfate, continue to centrifuge 5min under the conditions of 4000r/min after being homogenized 2min.Take supernatant 10mL extremely
Than in straight tube, nitrogen is blown to dry under 40 DEG C of water-baths, adds 2mL50% methanol aqueous solutions and redissolve, be vortexed after mixing and add 50mgPSA
Powder, 50mg anhydrous magnesium sulfates and 50mgC18Powder, is vortexed again for crossing 0.22 μm of filter membrane after mixing, obtains test sample.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
A chromatographic conditions
Chromatographic column：Using Agilent SB-C18Column (2.1 × 100mm, 1.8 μm, USA)
Column temperature：35℃；
Sampling volume：5μL；Flow velocity：0.3mL/min；
Mobile phase A：0.1% formic acid water；Mobile phase B：Acetonitrile=90~10:10~90 (volume ratios), gradient elution 0~
3min, A become 50% by 90；3~5min, A are changed into 10% from 50；It is 10% that A is maintained during 5~8min；8~10min, A are by 10
It is changed into 90%.
B Mass Spectrometry Conditions
Ion gun：Electric spray ion source (ESI)；Scan mode：Cation；
Detection mode：Multiple-reaction monitoring (MRM) mode gathers；
Detector：The triple level Four bar mass spectrographs of Agilent 6460 are furnished with ESI ion guns；Wherein qualitative ion pair, quantitative
The parameters such as ion pair, collision energy refer to table 1.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain total ion chromatogram (such as Fig. 4 institutes
Show).It is compared by instrument software comparison chart 1, Fig. 4, shows to be not detected by 34 kinds of pesticide residues in cucumber test sample.
It is remaining containing measurement for pesticide in the cucumber sample of 34 kinds of pesticide residues of 5.0 μ g/kg that embodiment 5 adds concentration
Surely comprise the following steps：
(1) test sample is prepared
Accurate to weigh through smashing the negative cucumber sample 5.00g of processing in example 2, addition concentration is in the sample
The hybrid working solution 1.0mL of 100.0 ng/mL, then the concentration of 35 kinds of pesticide residues is 5.0 μ g/kg in the apple sample,
Subsequent treatment obtains test sample with embodiment 4.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
With embodiment 4.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain total ion chromatogram (such as Fig. 5 institutes
Show).Sample quality according to representated by test sample, 34 kinds of agricultures in test sample are calculated from the standard curve of embodiment 1
The content of medicine residue.34 kinds of object contents in cucumber sample are calculated in the range of 4.0~5.2 μ g/kg, the rate of recovery
Within 80%~120%；Prove that the testing result of embodiment 4 is accurate.
The measure of 34 kinds of pesticide residue contents in 6 strawberry sample of embodiment
Comprise the following steps：
(1) test sample is prepared
It is accurate to weigh the strawberry sample 5.00g for smashing processing, 10mL extractants are added, are separately added into after homogeneous homogenate 1min
2 g sodium chloride and anhydrous magnesium sulfate, continue to centrifuge 5min under the conditions of 4000r/min after being homogenized 2min.Take supernatant 10mL extremely
Than in straight tube, nitrogen is blown to dry under 40 DEG C of water-baths, adds 2mL50% methanol aqueous solutions and redissolve, be vortexed after mixing and add 50mgPSA
Powder, 50mg anhydrous magnesium sulfates, 50mg florisil silicas and 50mgC18Powder, is vortexed again for crossing 0.22 μm of filter membrane after mixing,
Obtain test sample.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
A chromatographic conditions
Chromatographic column：Using Agilent SB-C18Column (2.1 × 100mm, 1.8 μm, USA)
Column temperature：35℃；
Sampling volume：5μL；Flow velocity：0.3mL/min；
Mobile phase A：0.1% formic acid water；Mobile phase B：Acetonitrile=90~10:10~90 (volume ratios), gradient elution 0~
3min, A become 50% by 90；3~5min, A are changed into 10% from 50；It is 10% that A is maintained during 5~8min；8~10min, A are by 10
It is changed into 90%..
B Mass Spectrometry Conditions
Ion gun：Electric spray ion source (ESI)；Scan mode：Cation；
Detection mode：Multiple-reaction monitoring (MRM) mode gathers；
Detector：The triple level Four bar mass spectrographs of Agilent 6460 are furnished with ESI ion guns；Wherein qualitative ion pair, quantitative
The parameters such as ion pair, collision energy refer to table 1.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain total ion chromatogram (such as Fig. 6 institutes
Show).It is compared by instrument software comparison chart 1, Fig. 6, shows to be not detected by 34 kinds of pesticide residues in strawberry test sample.
Embodiment 7 adds concentration as the remaining assay of pesticide in the strawberry sample of 34 kinds of pesticide residues of 5.0 μ g/kg
Comprise the following steps：
(1) test sample is prepared
Accurate to weigh through smashing the negative strawberry sample 5.00g of processing in example 2, addition concentration is in the sample
The hybrid working solution 1.0mL of 100.0 ng/mL, then the concentration of 34 kinds of pesticide residues is 5.0 μ g/kg in the apple sample,
Subsequent treatment obtains test sample with embodiment 6.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
With embodiment 6.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain total ion chromatogram (such as Fig. 7 institutes
Show).Sample quality according to representated by test sample, 34 kinds of agricultures in test sample are calculated from the standard curve of embodiment 1
The content of medicine residue.34 kinds of object contents in strawberry sample are calculated in the range of 4.0~5.2 μ g/kg, the rate of recovery
Within 80%~120%；Prove that the testing result of embodiment 6 is accurate.
The measure of 34 kinds of pesticide residue contents in 8 tomato sample of embodiment
Comprise the following steps：
(1) test sample is prepared
The tomato sample 5.00g for smashing processing is accurately weighed, 10mL extractants is added, adds respectively after homogeneous homogenate 1min
Enter 2g sodium chloride and anhydrous magnesium sulfate, continue to centrifuge 5min under the conditions of 4000r/min after being homogenized 2min.Take 10 mL of supernatant
Redissolved to 2mL50% methanol aqueous solutions than in straight tube, nitrogen is blown to dry under 40 DEG C of water-baths, are added, be vortexed after mixing and add 50
MgPSA powder, 50mg anhydrous magnesium sulfates, 50mg florisil silicas and 50mgC18Powder, is vortexed again for crossing 0.22 μm after mixing
Filter membrane, obtains test sample.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
A chromatographic conditions
Chromatographic column：Using Agilent SB-C18Column (2.1 × 100mm, 1.8 μm, USA)
Column temperature：35℃；
Sampling volume：5μL；Flow velocity：0.3mL/min；
Mobile phase A：0.1% formic acid water；Mobile phase B：Acetonitrile=90~10:10~90 (volume ratios), gradient elution 0~
3min, A become 50% by 90；3~5min, A are changed into 10% from 50；It is 10% that A is maintained during 5~8min；8~10min, A are by 10
It is changed into 90%..
B Mass Spectrometry Conditions
Ion gun：Electric spray ion source (ESI)；Scan mode：Cation；
Detection mode：Multiple-reaction monitoring (MRM) mode gathers；
Detector：The triple level Four bar mass spectrographs of Agilent 6460 are furnished with ESI ion guns；Wherein qualitative ion pair, quantitative
The parameters such as ion pair, collision energy refer to table 1.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain total ion chromatogram (such as Fig. 8 institutes
Show).It is compared by instrument software comparison chart 1, Fig. 8, shows to be not detected by 34 kinds of pesticide residues in tomato test sample
Thing.
Embodiment 9 adds concentration as the remaining content of pesticide in the tomato sample of 34 kinds of pesticide residues of 5.0 μ g/kg
Measure comprises the following steps：
(1) test sample is prepared
Accurate to weigh through smashing the negative tomato sample 5.00g of processing in example 2, addition concentration is in the sample
The hybrid working solution 1.0mL of 100.0 ng/mL, then the concentration of 34 kinds of pesticide residues is 5.0 μ g/kg in the apple sample,
Subsequent treatment obtains test sample with embodiment 8.
(2) ultra performance liquid chromatography-mass spectrum, Mass Spectrometry Conditions
With embodiment 8.
(3) measure
HPLC-MS/MS detections are carried out to the test sample that sampling volume is 5 μ L, obtain selection total ion chromatogram (as schemed
Shown in 9).Sample quality according to representated by test sample, is calculated 34 in test sample from the standard curve of embodiment 1
The content of kind pesticide residue.34 kinds of object contents in tomato sample are calculated in the range of 4.0~5.2 μ g/kg,
The rate of recovery is within 80%~120%；Prove that the testing result of embodiment 8 is accurate.
10 rate of recovery of embodiment and Precision Experiment
According to the test limit of method, to the apple sample in embodiment 2, the cucumber sample in embodiment 4, in embodiment 6
Strawberry, the tomato in embodiment 8 adds standard solution in each sample, carries out 3 realities according to the additive amount in table 3
Test.Average recovery rate result sees table 3.
Average TIANZHU XINGNAO Capsul of the 3 34 kinds of pesticide residue materials of table in four kinds of samples
| Apple | Cucumber | Strawberry | Tomato | |
| Pymetrozine | 81.5 | 90.5 | 91.8 | 91.8 |
| Propamocarb | 85.9 | 85.5 | 87.6 | 87.6 |
| Carbendazim | 88.0 | 88.5 | 82.3 | 91.0 |
| Probenazole | 85.5 | 85.9 | 81.3 | 81.3 |
| Oxamyl | 87.7 | 88.0 | 105.2 | 105.2 |
| Methomyl | 93.8 | 114.3 | 88.7 | 85.1 |
| Spanon | 110.0 | 96.4 | 84.6 | 84.6 |
| Diacloden | 86.5 | 87.4 | 84.4 | 84.4 |
| Imidacloprid | 84.2 | 86.6 | 88.5 | 80.2 |
| Imidaclothiz | 81.7 | 86.5 | 82.1 | 82.1 |
| Acetamiprid | 86.3 | 89.5 | 83.7 | 83.7 |
| Thiacloprid | 92.1 | 98.2 | 91.0 | 92.0 |
| Aldicarb | 87.9 | 93.1 | 91.1 | 91.1 |
| Imazalil | 81.8 | 88.8 | 90.2 | 90.2 |
| Thiophanate-methyl | 87.6 | 87.9 | 88.5 | 83.3 |
| Carbofuran | 100.9 | 93.7 | 94.2 | 94.2 |
| Chloropyuril | 88.7 | 89.4 | 84.0 | 90.4 |
| Carbaryl | 85.2 | 92.3 | 101.7 | 80.3 |
| Demeton | 88.6 | 89.3 | 90.2 | 81.1 |
| Chem hoe | 85.8 | 99.6 | 104.2 | 82.4 |
| Paclobutrazol | 85.8 | 88.0 | 86.4 | 89.1 |
| Diethofencarb | 87.8 | 89.0 | 93.3 | 93.3 |
| Boscalid | 85.4 | 91.5 | 93.0 | 80.8 |
| Triazolone | 81.3 | 89.8 | 88.1 | 89.4 |
| Penconazole | 84.5 | 88.0 | 88.2 | 88.2 |
| Hexythiazox | 110.7 | 96.5 | 94.5 | 94.5 |
| Tebufenozide | 105.3 | 108.7 | 84.0 | 84.0 |
| Cyazofamid | 97.9 | 91.2 | 86.1 | 86.1 |
| Zoxamide | 83.6 | 93.3 | 87.0 | 87.0 |
| Indoxacarb | 100.2 | 93.4 | 94.9 | 94.9 |
| Diazinon | 84.0 | 87.6 | 94.4 | 84.5 |
| Haloxyfop-P-methyl | 97.7 | 97.0 | 89.3 | 89.3 |
| Diafenthiuron | 94.3 | 100.1 | 92.5 | 92.5 |
| Fenazaquin | 92.3 | 90.8 | 89.5 | 89.5 |
Table 3 is the results show that in 4 kinds of food substrates are tested, the recovery of standard addition highest of cucumber matrix, apple, strawberry and
Tomato matrix recovery of standard addition is good.
Embodiment described above only expresses the several embodiments of the present invention, its description is more specific and detailed, but simultaneously
Therefore it cannot be interpreted as limiting the scope of the claims volume of the present invention.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to what is originally delivered
Protection domain.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
1. the extractant of sample pre-treatments during a kind of detection for garden stuff pesticide residue, includes the raw material of following parts by weight：0-8 parts
Acetonitrile, 0-5 parts of acetone, 0-3 parts of 1- butyl -3- methylimidazole dicyan amine salt.
2. the extractant of sample pre-treatments during the detection according to claim 1 for garden stuff pesticide residue, it is characterised in that：
Mass ratio in the extractant between acetonitrile, acetone and 1- butyl -3- methylimidazole dicyan amine salt is 2-8：1-5：0-3.
3. the sample-pretreating method that extractant described in application claim 1 detects garden stuff pesticide residue, comprises the following steps：
1) fruits and vegetables homogenised sample to be measured is mixed with foregoing extractant, obtains the first mixture；
2) the first mixture obtained in step 1) is mixed with sodium chloride and anhydrous magnesium sulfate, obtains the second mixture；
3) the second mixture is centrifuged, takes supernatant；
4) take the supernatant in step 3), carry out nitrogen and be blown to dry, then add methanol solution and redissolve, obtain the 3rd mixture；
5) the 3rd mixture in step 4) is mixed with PSA, C18, Florey diatomite and magnesium sulfate, it is mixed obtains the 4th
Compound；
6) lysate of the 4th mixture in step 5) is taken, filtering with microporous membrane is carried out, obtains the sample for detection.
4. the sample-pretreating method according to claim 3 to garden stuff pesticide residue detection, it is characterised in that：The step
It is rapid 1) in the quality of fruits and vegetables homogenised sample to be measured be 0.1-10g, the quality of extractant is 0.1-20mL；In the step 2)
The mass ratio of sodium chloride and anhydrous magnesium sulfate is 1：1-5；The quality sum of sodium chloride, anhydrous magnesium sulfate, mixes with described first
The mass ratio of thing is 1：1-10；The mass fraction of methanol solution in the step 4) is 0.1-80%, methanol solution additive amount
For 0.1-5mL；The quality sum of PSA, C18, Florey diatomite and magnesium sulfate in the step 5), mix with the described 3rd
The mass ratio of thing is 1：5-20；Mass ratio between the PSA, C18, Florey diatomite and magnesium sulfate is 1：0-5：0-5：0-
5, the aperture of the miillpore filter in the step 6) is 0.22-0.45 μm.
5. the sample-pretreating method according to claim 4 to garden stuff pesticide residue detection, it is characterised in that：The step
It is rapid 1) in the quality of fruits and vegetables homogenised sample to be measured be 0.1-5g, the quality of extractant is 0.1-10mL.
6. the sample-pretreating method according to claim 5 to garden stuff pesticide residue detection, it is characterised in that：The step
It is rapid 2) in sodium chloride and anhydrous magnesium sulfate mass ratio be 1：3-5.
7. the sample-pretreating method according to claim 6 to garden stuff pesticide residue detection, it is characterised in that：The step
It is rapid 4) in the mass fraction of methanol solution be 30-60%.
8. the sample-pretreating method according to claim 7 to garden stuff pesticide residue detection, it is characterised in that：The step
It is rapid 5) in PSA, C18, Florey diatomite and magnesium sulfate mass ratio be 1：0.5-2：0.5-2：0.5-2：0.5-2.
9. quick determination method that is a kind of while measuring 34 kinds of pesticide residues in fruits and vegetables, follows the steps below：
1) standard curve is made
A variety of agriculture residual standard product are respectively placed in volumetric flask, add acetonitrile dissolving and constant volume, are configured to single mark storing solution；Take certain
Single mark storing solution of the residual compound of each agriculture of volume is mixed, and is added methanol aqueous solution and is diluted to middle interstitial fluid, then takes middle interstitial fluid
The hybrid standard working solution for being configured to a variety of concentration is diluted, injects on HPLC-MS/MS and measures, obtains Multiple Pesticides residual
The standard curve of compound；
2) sample to be tested is measured
Pre-treatment first is carried out to sample to be tested, then upper machine measure obtains the total ion chromatogram of sample to be tested；Then will detection
Obtained sample to be tested inspection data, the remaining concentration of pesticide in sample to be tested is calculated by standard curve.
10. the according to claim 9 pair of quick determination method for measuring 34 kinds of pesticide residues in fruits and vegetables at the same time, its feature exist
In：The method for carrying out pre-treatment in the step 2) to sample to be tested is as follows：
1) fruits and vegetables sample to be measured is mixed with extractant；
2) mixture obtained in step 1) is mixed with sodium chloride and anhydrous magnesium sulfate；
3) centrifugal treating is carried out to the mixture obtained in step 2), takes supernatant；
4) take the supernatant liquid nitrogen in step 3) to be blown to dry, then add methanol solution and redissolve；
5) mixture in step 4) is mixed with PSA, C18, Florey diatomite and magnesium sulfate；
6) take the lysate of the mixture in step 5) to carry out filtering with microporous membrane, obtain the sample for detection；
The extractant includes the raw material of following parts by weight：0-8 parts of acetonitrile, 0-5 parts of acetone, 0-3 parts of 1- butyl -3- first
Base imidazoles dicyan amine salt.
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