CN105628839A

CN105628839A - GC-Q-TOF/MS detection technology for 708 pesticide residues in leaf vegetables

Info

Publication number: CN105628839A
Application number: CN201510992529.0A
Authority: CN
Inventors: 庞国芳; 范春林; 李岩; 常巧英
Original assignee: Chinese Academy of Inspection and Quarantine CAIQ
Current assignee: Chinese Academy of Inspection and Quarantine CAIQ
Priority date: 2015-12-28
Filing date: 2015-12-28
Publication date: 2016-06-01

Abstract

The invention relates to the field of detection of pesticide residues and provides a detection method for screening 708 pesticide residues in leaf vegetables. In the GC-Q-TOF/MS first-class mode, a fragment ion full-scanning mass spectrum on the specified conditions of a pesticide standard, the chemical formula, the retention time, the fragment ion accurate mass number and the ion abundance ratio of pesticide are obtained, guided into PCDL software and related to corresponding pesticide information to form a database, and full-spectrum data of samples to be tested is determined on the identical set condition to obtain full-spectrum data. A sample measuring result is retrieved through a precise mass database, the detected pesticide meeting the identification basis is accurately identified, and pesticide residue varieties in leaf vegetables are obtained. The detection method built by applying the GC-Q-TOF/MS high resolution mass spectrum is used for detecting pesticide residues in leaf vegetables, specified pretreatment is preferentially combined, and the advantages that detection is rapid and high in throughput, precision and reliability are achieved. No standard is used as a contrast, and meanwhile 708 pesticides in the leaf vegetables are qualitatively detected and confirmed.

Description

708 kinds of pesticide residue GC-Q-TOF/MS detection techniques in a kind of leaf vegetables

Technical field

The present invention relates to the residual detection field of agriculture, apply, in particular to a kind of, the method for detecting that in GC-Q-TOF/MS examination leaf vegetables, 708 kinds of agricultures are residual.

Background technology

Global environment detection system/food item (GlobalEnvironmentMonitoringSystem is jointly established as far back as the World Health Organization (WHO) in 1976, food and agricultural organization (FAO) and United Nations Environment Programme (UNEP) (UNEP); GEMS/Food); it is intended to grasp member state's food contamination situation; understand contaminants in food intake; protection HUMAN HEALTH, promotes Trade Development. Now, countries in the world all rise to food safety the strategic position of national security. Pesticide Residue is one of food security standard, is also international trade access threshold. Meanwhile, the requirement of pesticide residue is presented kind and gets more and more, more and more stricter development trend of limiting the quantity, be also exactly that the Pesticide Residue threshold set up of international trade is more and more higher. Present European Union has formulated 162248 MRL standards of 839 kinds of agricultural chemicals, and the U.S. has formulated 39147 MRL standards of 351 kinds of agricultural chemicals, and Japan has formulated 51600 multinomial MRL standards of 579 kinds of agricultural chemicals, and China has issued 3650 MRL standards of 381 kinds of agricultural chemicals for 2014. At present, the uniform limit limitation generally adopted in the world is 10 �� g/kg. Therefore, the quick Detecting Pesticide technology of high-throughput is all called in food safety and international trade, and this provides opportunities and challenges also to undoubtedly vast pesticide residue analysis worker. In current numerous Analytical Techniques of Pesticide Residues, hydrolysis and condensation realizes the optimized analysis means that the many residuals of high-throughput detect fast.

Current pesticide residue analysis is main mainly with gas-chromatography, liquid chromatography, gas chromatography-mass spectrum and liquid chromatograph mass spectrography technology. All first these detection techniques need standard sample of pesticide comparison to carry out qualitative. Such as, the detection of 100 kinds of agricultural chemicals just is needed to prepare corresponding 100 kinds of standard sample of pesticide comparison, and the agricultural chemicals outside these 100 kinds all can be missed. In the real work of agricultural and veterinary chemicals results from residue tests room, hundreds of kinds of standard sample of pesticide all can not be laid in most laboratory, and its reason is that standard sample of pesticide is not only expensive, and validity period only 2-3, it is necessary to overlapping investment. The standing standard sample of pesticide of common laboratory only tens kinds, the pesticide species of its daily monitoring is also just only limited to these tens kinds, thus causes food safety monitoring leak.

Summary of the invention

In order to overcome above-mentioned defect, the present inventor team concentrates on studies through for many years, have developed the GC-Q-TOF/MS detection techniques of 708 kinds of pesticide residue in the accurate mass database of 708 kinds of agricultural chemicals and a kind of fruit vegetables, achieve and do not need standard control, in leaf vegetables 708 kinds of pesticide residue can be carried out rapid detection detection, meet the urgent need that in current agricultural-food, pesticide residue high-throughput detects fast.

On the one hand, the present invention provides the method for detecting that a kind of GC-Q-TOF/MS detects 708 kinds of pesticide residue in leaf vegetables, it is characterised in that: set up 708 kinds of agricultural chemicals TOF/MS databases, in leaf vegetables 708 kinds of agricultural chemicals are carried out qualitative detecting and confirmation simultaneously; The method comprises the following steps:

(1) standard sample of pesticide is mixed with respectively the standardized solution of 1-5mg/L, measure the full modal data of often kind of standardized solution when setting through the full modal data of GC-QTOF/MS first class mode, obtain the chemical molecular formula of often kind of agricultural chemicals, retention time, fragmention accurate mass number, ion abundance ratio respectively;

(2) the full modal data of first class mode that will obtain in step (1), fragmention full scan mass spectrum after examining imports in PCDL software, and be associated with corresponding agricultural chemicals information, build up TOF/MS accurate mass database, for data retrieval analysis;

(3) the full modal data of agricultural chemicals in the mensuration testing sample of leaf vegetables when identical with step (1) setting, obtains the chemical molecular formula of its agricultural chemicals contained, retention time, fragmention accurate mass number, ion abundance ratio;

(4) to sample determination result through accurate mass database retrieval, the detection agricultural chemicals meeting appraisal basis is accurately identified, thus obtain the residual kind of the agriculture in leafy vegetable.

Preferably, by leaf vegetables carrying out pre-treatment to obtain the mensuration testing sample of leaf vegetables, described pre-treating process comprises following program:

Prepared by sample: leaf vegetables sample extracts through 0.5-5 volume % acetate acetonitrile homogenate, after dewatering and be centrifugal, concentrated, then through Carbon/NH₂Column purification, acetonitrile+toluene wash-out remains of pesticide, makes after reconcentration, filtration and treats sample measuring liquid a.

The useful effect that the present invention has:

1, the present invention is taking compound information such as high resolution fragmention accurate mass number, retention time, ion abundance ratios as identifying standard, the thousand kinds of agricultural chemicals detectings set up and confirmation technological method, revolutionize original taking compound standard substance as joining the qualitative model of ratio, it is that one does not need standard substance to compare, quick, high-throughput, accurately and reliably Detecting Pesticide new technology.

2, the present invention is under the prerequisite not needing standard substance to compare, it is achieved that the qualitative detecting of 708 kinds of pesticide residue and confirmation, it is possible to greatly save the cost buying standard substance, also environmental protection more, safety.

3, the present invention can complete 708 kinds of agricultural chemicals detecting, qualitative with confirmation in 1 hour, compared with traditional detection method, it is possible to increase work efficiency hundreds of times.

4, the present invention can accomplish 708 kinds of disposable extraction and cleanings of pesticide residue in leaf vegetables sample, and single injected sampling detects, compared with traditional detection method, it is possible to saves testing cost, increase work efficiency hundreds of times.

5, leaf vegetables sample middle peasant's cartridge bag that the present invention can test draws together the class in following material or a few class: organophosphorus insecticide, carbamate chemicals for agriculture, Ion pairing, sulfonylurea pesticide, nicotinoids agricultural chemicals, the agricultural chemicals of cinerins agricultural chemicals and other types and metabolite.

6, the GC-QTOF/MS detection techniques index that the present invention sets up: sweep limit 50-600m/z; The accurate mass measured can reach 0.0001m/z; Within Mass accuracy can be controlled in 20ppm; Sweep velocity is 5 times every second in 50-600m/z scope.

7, in one application gas-chromatography level Four bar flight time mass spectrum (GC-Q-TOF/MS) rapid screening leaf vegetables provided by the invention, the high resolution mass spectrum detection techniques of 708 kinds of pesticide residue obtains promotion and application in 7 pesticide residue analysis laboratories, through the detecting of the 30 multiple commercially available leaf vegetables samples of Duo Ge provinces and cities being verified, achieve important residual detecting data.

8, in the 708 kinds of agricultural chemicals simultaneously detected, detecting sensitivity without exception lower than standard 10 microgram/kilogram, meet the requirement of the horizontal examination of various countries pesticide residue MRL.

Other features and advantages of the present invention are described in detail in embodiment part subsequently.

Accompanying drawing explanation

Accompanying drawing is used to provide a further understanding of the present invention, and forms a part for specification sheets, is used from explanation the present invention with embodiment one below, but is not construed as limiting the invention. In the accompanying drawings:

Fig. 1 is commercially available naked oats dish sample GC-Q-TOF/MS technology frequency the highest agricultural chemicals (pyridaben) result example.

Fig. 2 is the result compared in the mass spectrum information and date storehouse of the highest agricultural chemicals of the frequency (pyridaben) of commercially available naked oats dish sample GC-Q-TOF/MS technology detection.

Fig. 3 is commercially available celery sample GC-Q-TOF/MS technology frequency the highest agricultural chemicals (carbofuran) result example.

Fig. 4 is the result compared in the mass spectrum information and date storehouse of the highest agricultural chemicals of the frequency (carbofuran) of commercially available celery sample GC-Q-TOF/MS technology detection.

Fig. 5 is commercially available rape sample GC-Q-TOF/MS technology frequency the highest agricultural chemicals (pyridaben) result example

Fig. 6 is the result compared in the mass spectrum information and date storehouse of the highest agricultural chemicals of the frequency (pyridaben) of commercially available rape sample GC-Q-TOF/MS technology detection.

Fig. 7 is commercially available romaine lettuce sample GC-Q-TOF/MS technology frequency the highest agricultural chemicals (metaxanin) result example

Fig. 8 is the result compared in the mass spectrum information and date storehouse of the highest agricultural chemicals of the frequency (metaxanin) of commercially available romaine lettuce sample GC-Q-TOF/MS technology detection.

Fig. 9 is commercially available water spinach sample GC-Q-TOF/MS technology frequency the highest agricultural chemicals (procymidone) result example.

Figure 10 is the result compared in the mass spectrum information and date storehouse of the highest agricultural chemicals of the frequency (procymidone) of commercially available water spinach sample GC-Q-TOF/MS technology detection.

Embodiment

Hereinafter the specific embodiment of the present invention is described in detail. Should be understood that, embodiment described herein, only for instruction and explanation of the present invention, is not limited to the present invention.

The present invention provides the method for detecting that a kind of GC-Q-TOF/MS detects 708 kinds of pesticide residue in leaf vegetables, it is characterised in that: set up 708 kinds of agricultural chemicals TOF/MS databases, in leaf vegetables 708 kinds of agricultural chemicals are carried out qualitative detecting and confirmation simultaneously; The method comprises the following steps:

Concrete, illustrate that the GC-QTOF/MS database of the present invention is set up by following method for Tsidial (Phenthoate):

1) at data acquisition interface, Tsidial (Phenthoate) standard substance (1-5mg/L) is carried out fragmention full scan mass spectrum collection through GC-QTOF/MS first class mode under appointment chromatogram Mass Spectrometry Conditions.

Chromatographic condition is: chromatographic column: VF-1701MS (30m �� 0.25mm �� 0.25um) (J&WScientific, Folsom, CA); Chromatographic column heating schedule: 30-50 DEG C keeps 0.5-5min, then to be warming up to 200-280 DEG C with 3-10 DEG C/min again, then it is warming up to 110-150 DEG C with 20-50 DEG C/min, it is warming up to 200-280 DEG C again with 3-10 DEG C/min, it is warming up to 250-320 DEG C subsequently with 5-15 DEG C/min, keep 3-10min, then it is cooled to 200-290 DEG C with 5-20 DEG C/min, keep 1-5min; Carrier gas: helium; Purity 99.999%; 0.5-2mL/min; Injector temperature: 230-320 DEG C; Sample size 1 �� L; Input mode: Splitless injecting-Sample, solvent delay 2-10min. Preferably, chromatographic condition: chromatographic column: VF-1701MS (30m �� 0.25mm �� 0.25um) (J&WScientific, Folsom, CA); Chromatographic column heating schedule: 40 DEG C keep 1min, are then warming up to 130 DEG C with 30 DEG C/min, then are warming up to 250 DEG C with 5 DEG C/min, be warming up to 300 DEG C with 10 DEG C/min subsequently, keep 5min, then are cooled to 280 DEG C with 10 DEG C/min, keep 2min; Carrier gas: helium, purity 99.999%; Flow velocity: 1.2mL/min; Injector temperature: 290 DEG C; Sample size 1 �� L; Input mode: Splitless injecting-Sample, solvent delay 6min.

Mass Spectrometry Conditions is: ion source: EI (electron impact ionization source); Ion source temperature: 230-330 DEG C, electron bombardment ionization source energy: 50-100eV; Gas phase-mass spectrometer interface temperature: 200-320 DEG C; Scan mode: carry out TOF pattern full scan in the way of every second 2-10 opens spectrogram, total mass number scope 50-600amu. Preferably, Mass Spectrometry Conditions: ion source: EI (electron impact ionization source); Ion source temperature: 280 DEG C, electron bombardment ionization source energy: 70eV; Gas phase-mass spectrometer interface temperature: 290 DEG C; Scan mode: carried out TOF pattern full scan by every second in the way of 5 spectrograms, total mass number scope 50��600amu..

Described fragmention full scan mass spectrum collection is the mass spectrum information of Tsidial (Phenthoate) standard substance, comprises compound chemical molecular formula, retention time, fragmention accurate mass number, ion abundance ratio.

2) in the qualitative software of AgilentMassHunter (instrument carries), invocation target agricultural chemicals (Phenthoate) gathers file, and determines target agricultural chemicals chromatographic peak.

3) after opening target agricultural chemicals mass spectrum deduction background, it may also be useful to low resolutions of NIST11 is composed storehouse (instrument carries) and carried out searching for the information obtaining compound, and verify library searching to compound whether be consistent with actual.

4) mass spectrum is verified, target compound mass spectrum can show composes, through low resolution of NIST11, fragmention molecular formula and the accurate mass number that storehouse matches, by " EditPeakAnnotion " window, ion information is carried out the amendment of necessity, the ion that such as accurate mass number deviation is excessive, may mean that the Formula structure that software calculates automatically is wrong, now manually can input correct information in " Formula " arranges, or " IronType " is changed to Unknown.

5) fragmention full scan mass spectrum after examining imports in PCDL software (instrument carries), and is associated with Tsidial information, builds up TOF/MS database. In database, the mass spectrum information of 708 kinds of agricultural chemicals, refers to subordinate list 1.

Preferably, described agricultural chemicals is 708 shown in table a kind agricultural chemicals. The method forming these agricultural chemicals can carry out according to the mode of the TOF/MS database as above forming Tsidial (Phenthoate).

According to the present invention, the mass spectrum information of the standard substance obtained can being carried out according to PCDL software manual book by the method that PCDL software and corresponding agricultural chemicals are interrelated, the present invention does not repeat them here.

According to the present invention, described fruit vegetables can be various fruit and vegetables, and the present invention is to this not special restriction. Preferably, described fruit leaf vegetables is selected from the group being made up of celery, water spinach, naked oats dish, crowndaisy chrysanthemum, Chinese cabbage, Plantula Brassicae chinensis, spinach, leaf mustard, the certain herbaceous plants with big flowers that falls, three-coloured amaranth, pachyrhizus leaf, rape, hare's-lettuce, green vegetables, taro, asparagus lettuce, purple back of the body dish, baby cabbage, Leaf of Matrimonyvine, spring vegetable, youngster's dish, milk Chinese cabbage, romaine lettuce, crowndaisy chrysanthemum, great Ye crowndaisy chrysanthemum, Leaf lettuce, head lettuce, corn salad, Folium Raphani, turnip leaf, witloof, sweet fennel and Corm Fructus Foeniculi etc.

Table 1

According to the present invention, with the full modal data of agricultural chemicals in the mensuration testing sample of leaf vegetables when setting up identical setting, obtain the chemical molecular formula of its agricultural chemicals contained, retention time, fragmention accurate mass number, ion abundance ratio; Call the TOF/MS database set up, according to TOF/MS database to the qualitative retrieval of target compound, search argument retention time deviation is defined as �� 0.15min, accurate mass deviation is defined as �� 20ppm, and software can provide the retrieval matching score value of each detection ion automatically according to the deviation of theoretical value in the measured value of the key elements such as often kind of compound retention time, accurate mass and TOF/MS database. Detection ion score mean value is this compound integrate score, specify each compound score value > 60 and at least three ion detections, it is positive detection, according to the method accuracy of pesticide residue in the qualitative detecting agricultural-food of TOF/MS database, reaches more than 90%.

According to the present invention, by leaf vegetables carrying out pre-treatment to obtain the mensuration testing sample of leaf vegetables, described pre-treating process preferably includes following program:

Fruit vegetables sample is extracted through acetonitrile solution (the being called for short 0.5-5 volume % acetate acetonitrile) homogenate containing 0.5-5 volume % acetic acid, after dewatering and be centrifugal, concentrated, then through Carbon/NH₂Column purification, acetonitrile+toluene wash-out remains of pesticide, makes after reconcentration, filtration and treats sample measuring liquid;

Wherein, it is preferable that, described leaf vegetables sample size is 5-20g, and the consumption that the consumption of the acetate acetonitrile of described 0.5-5 volume % is 30-50mL is preferably 35-45ml; And the fruit vegetables sample adding acetate acetonitrile is placed in centrifuge tube and carries out homogenized. Described homogenate extraction conditions is preferably: rotating speed 12000-15000rpm, time 0.5-2min.

Wherein, described through dewatering preferably by the sodium-chlor adding 0.5-5g in acetate acetonitrile extracting solution, the anhydrous magnesium sulfate of 2-8g, and the 4-6min that vibrates realizes. Then the dehydrated sample centrifugal 1-10min under 4000-4500rpm that will obtain, obtains supernatant liquor, and concentrates the supernatant liquor 10-30mL obtained rotary evaporation in the water-bath of 20-60 DEG C to 0.5-2mL.

Wherein, described Carbon/NH₂Column purification step is preferably as follows: at Carbon/NH₂Post adds the preferred 2cm high anhydrous sodium sulphate of about 1-3cm. First with 2-6mL preferred 4mL acetonitrile+toluene (preferred 2.5-3.5:1, it is more preferable to be 3:1, v/v) drip washing SPE post, and abandon effluent liquid, when liquid level arrive sodium sulfate top time, rapidly sample concentration liquid is transferred on decontaminating column, under connect new heart bottle receive. Each 2-6mL preferred 2mL acetonitrile+toluene (preferred 2.5-3.5:1, it is more preferable to be 3:1, v/v) washs sample liquid bottle three times again, and is moved in SPE post by washings. Again with 20-30mL preferred 25mL acetonitrile+toluene (preferred 2.5-3.5:1, it is more preferable to be 3:1, v/v) wash-out agricultural chemicals, merge elutriant in heart bottle.

Wherein, described treat that sample measuring liquid is prepared preferably by such as under type further, the elutriant spin concentration extremely about 0.5-2mL in 30-50 DEG C of water-bath that will as above obtain, and concentrated solution is placed in nitrogen blowing down and does, acetonitrile+0-5% formic acid the water (2+ (7-9), it is preferable that 2+8, v/v) adding 1-5mL is mixed even, after 0.18-0.22 ��m preferably 0.2 ��m of membrane filtration, fixed appearance, obtains treating sample measuring liquid.

The specific embodiment of the present invention below will be described in detail.

Embodiment 1

The present embodiment is for illustration of the residual method for detecting of 708 kinds of agricultures in GC-Q-TOF/MS examination leaf vegetables provided by the invention

(1) area naked oats dish sample, 10g Chengdu (being accurate to 0.01g) is taken, in 80ml centrifuge tube, add the acetonitrile solution of 40mL1% acetic acid, with high-speed homogenization machine 13500r/min, 1min is extracted in homogenate, add 1g sodium-chlor, 4g anhydrous magnesium sulfate, vibration 5min, centrifugal 5min under 4200r/min, getting supernatant liquor 20mL, in 40 DEG C of water-baths, rotary evaporation concentrates to about 1mL, to be clean.

(2) at Carbon/NH₂Post adds about 2cm high anhydrous sodium sulphate. First with 4mL acetonitrile+toluene (3+1, v/v) drip washing SPE post, and abandon effluent liquid, when liquid level arrive sodium sulfate top time, rapidly sample concentration liquid is transferred in SPE post, under connect new heart bottle receive. Wash sample liquid bottle three times with 2mL acetonitrile+toluene (3+1, v/v) more every time, and washings is moved in SPE post. Post connects 50mL liquid vessel, with 25mL acetonitrile+toluene (3+1, v/v) wash-out agricultural chemicals and related chemistry product, is incorporated in heart bottle, and spin concentration is to about 0.5mL in 40 DEG C of water-baths.

(3) concentrated solution is placed in nitrogen blowing down and does, and the acetonitrile+water (2+8, v/v) adding 2mL is mixed even, fixed after 0.2 ��m of membrane filtration holds, and obtains testing sample solution.

(4) GC-Q-TOF/MS operational condition, is separated and obtains the credit minor of each each fragmention of agricultural chemicals in naked oats dish sample, retention time, fragmention accurate mass number, ion abundance ratio.

Chromatographic condition: chromatographic column: VF-1701MS (30m �� 0.25mm �� 0.25um) (J&WScientific, Folsom, CA); Chromatographic column heating schedule: 40 DEG C keep 1min, are then warming up to 130 DEG C with 30 DEG C/min, then are warming up to 250 DEG C with 5 DEG C/min, be warming up to 300 DEG C with 10 DEG C/min subsequently, keep 5min, then are cooled to 280 DEG C with 10 DEG C/min, keep 2min; Carrier gas: helium, purity 99.999%; Flow velocity: 1.2mL/min; Injector temperature: 290 DEG C; Sample size 1 �� L; Input mode: Splitless injecting-Sample, solvent delay 6min.

Mass Spectrometry Conditions: Mass Spectrometry Conditions: ion source: EI (electron impact ionization source); Ion source temperature: 280 DEG C, electron bombardment ionization source energy: 70eV; Gas phase-mass spectrometer interface temperature: 290 DEG C; Scan mode: carried out TOF pattern full scan by every second in the way of 5 spectrograms, total mass number scope 50��600amu..

(5) database of foundation table 1 is to the qualitative retrieval of acquisition information in step (4), search argument retention time deviation is defined as �� 0.5min, accurate mass deviation is defined as �� 100ppm, detection at least three fragmentions, and the deviation according to theoretical value in the measured value of often kind of each key element of compound and table 1 database, automatically provide retrieval matching score value, the compound of retrieval matching score value > 60, namely confirm this target compound of detection. Detected result is in table 2.

For the pyridaben that frequency is higher, as shown in Figure 1, naked oats dish detects out 7 fragmention information, with database retrieval, with pyridaben comparison, 7 fragmention molecular weight deviation marks are all more than 87 points, retention time deviation is within 0.003min, the fragment ion masses deviation that abundance is the highest is-3.58ppm (as shown in Figure 2), it is possible to confirms, has pyridaben to examine in naked oats dish.

Embodiment 2

Detect with the agriculture in batch naked oats dish sample is residual according to the method in embodiment 1, the difference is that, pre-treatment step is as follows:

(1) accurately taking 10.00g sample in centrifuge tube, add 50mL water, vibration is until sample is fully infiltrated by water. 50mL acetonitrile is pipetted to, in centrifuge tube, being then placed in by centrifuge tube on the whirlpool mixing vibration instrument of whirlpool, with 2000rpm rate oscillation 1min after freezing 10min. 10min is kept under centrifuge tube is placed in-10 DEG C of conditions, then in centrifuge tube, add 20g anhydrous magnesium sulfate and 5g sodium-chlor, 5g Trisodium Citrate and 2.5g Sodium citrate, immediately on whirlpool mixing vibration instrument, with 2000rpm rate oscillation 2min, then with the centrifugal 10min of 4000rpm speed, supernatant liquor is obtained.

(2) pipette supernatant liquor 1.0mL in 1.5mL centrifuge tube, add the PSA sorbent material of 150mg anhydrous magnesium sulfate and 25mg, with 2000rpm rate oscillation 2min on whirlpool mixing vibration instrument, with the centrifugal 2min of 6000rpm speed. Aspirate supernatant, through 0.22 ��m of organic phase membrane filtration, pipettes 200 �� L, adds 100 �� L acetonitriles, is diluted to 1.0mL with ultrapure water, obtains testing sample solution.

Detected result is in table 2.

Table 2

By above data it may be seen that adopt the method for the present invention, especially preferred method more effective can detect out the residual kind of agricultural-food middle peasant, and has lower detectability.

Embodiment 3

Detect the agriculture in leafy vegetable is residual according to the method in embodiment 1, the difference is that, sample is area, Chengdu celery sample.

Gathering 11, the commercially available celery sample in area, Chengdu, detection 19 kinds of pesticide residue, amount to 34 frequencys, it relates to 11, sample, concrete outcome is in table 3.

Table 3

For the carbofuran that frequency is higher: as shown in Figure 3, celery detects out 6 fragmention information, with database retrieval, with carbofuran comparison, 6 fragmention coupling marks are all more than 91 points, and retention time deviation is within 0.01min, and the fragment ion masses deviation that abundance is the highest is 8.82ppm (as shown in Figure 4), can confirm that celery has carbofuran detection.

Embodiment 4

Detect the agriculture in leafy vegetable is residual according to the method in embodiment 1, the difference is that, sample is degree area rape sample.

Gathering 8, the commercially available rape sample in area, Chengdu, detection 11 kinds of pesticide residue, amount to 20 frequencys, it relates to 8, sample, concrete outcome is in table 4.

Table 4

For the pyridaben that frequency is higher: as shown in Figure 5, rape detects out 7 fragmention information, with database retrieval, with pyridaben comparison, 7 fragmention molecular weight deviation marks are all more than 99 points, and retention time deviation is 0min, and the fragment ion masses deviation that abundance is the highest is-16.24ppm (as shown in Figure 6), can confirm that rape has pyridaben detection.

Embodiment 5

Detect the agriculture in leafy vegetable is residual according to the method in embodiment 1, the difference is that, sample is degree area romaine lettuce sample.

Gathering 14, the commercially available romaine lettuce sample in area, Chengdu, detection 14 kinds of pesticide residue, amount to 34 frequencys, it relates to 14, sample, concrete outcome is in table 5.

Table 5

For the metaxanin that frequency is higher: as shown in Figure 7, romaine lettuce detects out 6 fragmention information, with database retrieval, with metaxanin comparison, 6 fragmention molecular weight coupling marks are all more than 96 points, and retention time deviation is within 0.007min, and the fragment ion masses deviation that abundance is the highest is-20.72ppm (as shown in Figure 8), can confirm that romaine lettuce has metaxanin detection.

Embodiment 6

Detect the agriculture in leafy vegetable is residual according to the method in embodiment 1, the difference is that, sample is degree area water spinach sample.

Gathering 9, the commercially available water spinach sample in area, Chengdu, detection 6 kinds of pesticide residue, amount to 9 frequencys, it relates to 6, sample, concrete outcome is in table 6.

Table 6

For the procymidone that frequency is higher: as shown in Figure 9, water spinach detects out 6 fragmention information, with database retrieval, contrast with procymidone, 6 fragmention molecular weight coupling marks are all more than 95 points, and retention time deviation is within 0.007min, and the fragment ion masses deviation that abundance is the highest is 33.1ppm (as shown in Figure 10), can confirm that water spinach has procymidone detection.

Below the preferred embodiment of the present invention is described in detail; but, the detail that the present invention is not limited in above-mentioned enforcement mode, within the scope of the technical conceive of the present invention; the technical scheme of the present invention can being carried out multiple simple variant, these simple variant all belong to protection scope of the present invention.

It should be noted that in addition, each the concrete technology feature described in above-mentioned embodiment, when not contradiction, it is possible to combined by any suitable mode. In order to avoid unnecessary repetition, various possible array mode is illustrated by the present invention no longer separately.

In addition, can also carrying out arbitrary combination between the various different enforcement mode of the present invention, as long as it does not run counter to the thought of the present invention, it should be considered as content disclosed in this invention equally.

Claims

1. a GC-Q-TOF/MS detects the method for detecting of 708 kinds of pesticide residue in leaf vegetables, it is characterised in that: set up 708 kinds of agricultural chemicals TOF/MS databases, in leaf vegetables 708 kinds of agricultural chemicals are carried out qualitative detecting and confirmation simultaneously; The method comprises the following steps:

2. method according to claim 1, wherein, described agricultural chemicals is 708 shown in table a kind agricultural chemicals.

3. method according to claim 1, wherein, described leaf vegetables is preferably selected from the group being made up of celery, water spinach, naked oats dish, crowndaisy chrysanthemum, Chinese cabbage, Plantula Brassicae chinensis, spinach, leaf mustard, the certain herbaceous plants with big flowers that falls, three-coloured amaranth, pachyrhizus leaf, rape, hare's-lettuce, green vegetables, taro, asparagus lettuce, purple back of the body dish, baby cabbage, Leaf of Matrimonyvine, spring vegetable, youngster's dish, milk Chinese cabbage, romaine lettuce, crowndaisy chrysanthemum, great Ye crowndaisy chrysanthemum, Leaf lettuce, head lettuce, corn salad, Folium Raphani, turnip leaf, witloof, sweet fennel and Corm Fructus Foeniculi.

4. method according to claim 1, wherein, the condition measuring the full modal data of agricultural chemicals in often kind of standardized solution comprises:

(1) chromatographic condition: chromatographic column: VF-1701MS; Chromatographic column heating schedule: 30-50 DEG C keeps 0.5-5min, then it is warming up to 110-150 DEG C with 20-50 DEG C/min, it is warming up to 200-280 DEG C again with 3-10 DEG C/min, it is warming up to 250-320 DEG C subsequently with 5-15 DEG C/min, keep 3-10min, it is cooled to 200-290 DEG C with 5-20 DEG C/min again, keeps 1-5min; Carrier gas: helium; Flow velocity: 0.5-2mL/min; Injector temperature: 230-320 DEG C: Splitless injecting-Sample, solvent delay 2-10min;

(2) Mass Spectrometry Conditions: ion source: EI; Ion source temperature: 230-330 DEG C, electron bombardment ionization source energy: 50-100eV; Gas phase-mass spectrometer interface temperature: 200-320 DEG C; Scan mode: carry out TOF pattern full scan in the way of every second 2-10 opens spectrogram, total mass number scope 50-600amu.

5. method according to claim 1, wherein, TOF/MS accurate mass database can will measure the measured value of the retention time of the fragmention obtained, accurate mass number, ion abundance ratio from testing sample, with the retrieval comparison of the corresponding key element theoretical value of fragmention in TOF/MS accurate mass database, provide the score of each fragmention, and then provide the matching degree score value of average mark and target compound; Wherein, the retention time deviation setting of fragmention is �� 100ppm at �� 0.5min, accurate mass number deviation setting.

6. method according to claim 5, wherein: carry out the detection agricultural chemicals meeting appraisal basis by the following method accurately identifying:

By to characteristic ion detection situation and degree of conformity, the score value provided, agricultural chemicals is carried out qualitative recognition, its method program is: for the detecting of target compound in sample, it it is the result of the retention time of the compound that flight time mass spectrum is recorded, 7 fragmention accurate mass numbers that abundance is the highest, fragmention relative abundance, with TOF/MS database retrieval comparison, result is detection at least three fragmentions, and the agricultural chemicals that integrate score is greater than 60 is confirmed as detection agricultural chemicals.

7. method according to claim 1, by leaf vegetables carrying out pre-treatment to obtain the mensuration testing sample of leaf vegetables, described pre-treating process comprises following program:

8. method according to claim 7, wherein: described leaf vegetables sample size is 5-20g;

Preferably, the consumption of the acetate acetonitrile of described 0.5-5 volume % is 30-50mL;

Preferably, described homogenate extraction conditions is: rotating speed 12000-15000r/min, and the time is 0.5-5min;

Preferably, described referring to through dehydration adds 0.5-5g sodium-chlor, 2-8g anhydrous magnesium sulfate in 0.5-5 volume % acetate acetonitrile extracting solution, vibration 1-10min;

Preferably, described centrifugal centrifugal 1-10min under 3000-5000r/min is referred to;

Preferably, described concentrated referring to gets centrifuged supernatant 10-30mL, and in 20-60 DEG C of water-bath, rotary evaporation concentrates to about 0.5-2mL.

9. method according to claim 8, wherein: described Carbon/NH₂Column purification step is as follows: at Carbon/NH₂Post adds about 1-3cm high anhydrous sodium sulphate, first with a certain proportion of acetonitrile of 2-6mL and toluene mixing solutions drip washing SPE post, and abandons effluent liquid, when liquid level arrive sodium sulfate top time, rapidly sample concentration liquid is transferred on decontaminating column, under connect container reception; Wash sample liquid bottle three times with 2-6mL acetonitrile and toluene mixing solutions more every time, and washings is moved in SPE post. Again with the certain volume acetonitrile of preferred 20-30ml and toluene mixing solutions wash-out agricultural chemicals, merge elutriant in container;

Preferably, described a certain proportion of acetonitrile and toluene mixing solutions, acetonitrile: volume of toluene is than being 2.5-3.5:1.

10. method according to claim 8, wherein: described making after reconcentration, filtration treats that sample measuring liquid a refers to the elutriant that claim 9 obtains, in 30-50 DEG C of water-bath, spin concentration is to about 0.5-2mL, and concentrated solution is placed in nitrogen blowing down and does, the a certain proportion of acetonitrile and the 0-5 volume % formic acid water mixed solution that add 1-5mL mix even, after 0.18-0.22 ��m of membrane filtration, fixed appearance, obtains testing sample solution a.

Preferably, described a certain proportion of acetonitrile and 0-5 volume % formic acid water mixed solution, acetonitrile: 0.1-5 volume % formic acid water 2:7-9.