CN105588911A - GC-Q-TOF/MS detection technology for 708 pesticide residues in bud vegetables - Google Patents
GC-Q-TOF/MS detection technology for 708 pesticide residues in bud vegetables Download PDFInfo
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Abstract
The invention relates to the field of detection of pesticide residues and provides a detection method for screening 708 pesticide residues in bud vegetables. Fragment ion full-scan mass spectrograms under pesticide standard specified conditions are acquired in the GC-Q-TOF/MS level-one mode, chemical formulas, retention time, fragment ion accurate mass numbers and ion abundance ratios of pesticides are obtained, imported into PCDL software and associated with corresponding pesticide information, a database is formed, full spectrum data of to-be-detected samples are detected under same set conditions, and the full spectrum data are obtained; detection results of the samples are retrieved in an accurate mass database, detected pesticides conforming to identification bases are accurately identified, and types of the pesticide residues in the bud vegetables are obtained. The pesticide residues in the bud vegetables are detected with the detection method established by the aid of a GC-Q-TOF/MS high-resolution mass spectrum, optimization is combined with specific pretreatment, and the method has the characteristics of quickness, high flux, high accuracy and high reliability of detection. A standard is not required to serve as a contrast, and qualitative detection and confirmation are performed on the 708 pesticide residues in the bud vegetables simultaneously.
Description
Technical field
The present invention relates to the residual detection field of agriculture, particularly in a kind of application GC-Q-TOF/MS examination sprouts of vegetable vegetables 708Plant the residual method for detecting of agriculture.
Background technology
Fruit and vegetable and people's life is closely bound up, especially sprouts of vegetable vegetables (mung bean sprouts, Chinese toon bud and toothed burclover etc.)Nutritive value enjoy consumer's favor. But, at present in the plantation of sprouts of vegetable vegetables, may in growth and preserving processThe agricultural chemicals and the chemicals that use have exceeded 1100 kinds. But, follow being widely used of agricultural chemicals, its excessive use, abuse and mistakeWith wait abnormal activity also to produce thereupon, make residues of pesticides in agricultural product, thereby cause potential threat to human health, withTime also the foreign trade of agricultural product has been produced to adverse effect.
Common as far back as the World Health Organization (WHO), food and agricultural organization (FAO) and United Nations Environment Programme (UNEP) in 1976Global environment detection system/food item (GlobalEnvironmentMonitoringSystem, GEMS/ are set upFood), be intended to grasp member state's food pollution situation, understand food contaminant intake, protection health, gives a impetus to trade and sends outExhibition. Now, countries in the world all rise to food security the strategic position of national security. Pesticide Residue is food security markOne of accurate, be also international trade access threshold. Meanwhile, the requirement of residues of pesticides is presented to kind more and more, limit the quantity more nextStricter development trend, the Pesticide Residue threshold that namely international trade is set up is more and more higher. European Union formulates now162248 MRL standards of 839 kinds of agricultural chemicals, the U.S. has formulated 39147 MRL standards of 351 kinds of agricultural chemicals, Japan has formulated51600 multinomial MRL standards of 579 kinds of agricultural chemicals, China has issued 3650 MRL standards of 381 kinds of agricultural chemicals for 2014. At present, stateThe uniform limit limitation generally adopting on border is 10 μ g/kg. Therefore, the quick agriculture of high flux is all called in food security and international tradeMedicine residue detection technology, this provides opportunities and challenges also to undoubtedly vast pesticide residue analysis worker. Current numerousIn Analytical Techniques of Pesticide Residues, hydrolysis and condensation is the optimized analysis means that realize the how residual fast detecting of high flux.
Pesticide residue analysis is mainly with gas-chromatography, liquid chromatogram, gas chromatography-mass spectrum and liquid phase chromatogram-mass spectrometry combination at presentBe main by technology. First these detection techniques all need standard sample of pesticide contrast to carry out qualitative. For example, the inspection to 100 kinds of agricultural chemicalsSurvey and just need to prepare corresponding 100 kinds of standard sample of pesticide and contrast, and agricultural chemicals outside these 100 kinds all can be missed. At agricultural and veterinary chemicalsIn the real work of results from residue tests chamber, hundreds of standard sample of pesticide can not laid in most laboratories, and its reason is agricultural chemicalsStandard items are not only expensive, and the term of validity only has 2-3, need overlapping investment. The standing standard sample of pesticide of common laboratoryOnly have tens kinds, the pesticide species of its daily monitoring is also just only limited to these tens kinds, causes thus food safety monitoring leak.
Summary of the invention
In order to overcome above-mentioned defect, inventor team, through concentrating on studies for many years, has researched and developed the accurate matter of 708 kinds of agricultural chemicalsIn amount database and a kind of fruits and vegetables, the GC-Q-TOF/MS detection techniques of 708 kinds of residues of pesticides, has realized and has not needed standard items pairAccording to, can carry out rapid detection detection to 708 kinds of residues of pesticides in sprouts of vegetable vegetables, met Agricultural Products Pesticide ResiduesThe urgent need of high flux fast detecting.
On the one hand, the invention provides the detecting of 708 kinds of residues of pesticides in a kind of GC-Q-TOF/MS detection sprouts of vegetable vegetablesMethod, is characterized in that: set up 708 kinds of agricultural chemicals TOF/MS databases, carry out qualitative simultaneously to 708 kinds of agricultural chemicals in sprouts of vegetable vegetablesDetecting and confirmation; The method comprises the following steps:
(1) standard sample of pesticide is mixed with respectively to the standard liquid of 1-5mg/L, imposes a condition lower to GC-QTOF/MS one-levelPattern is composed the full spectrum data of every kind of standard liquid of data determination entirely, obtain respectively every kind of agricultural chemicals chemical molecular formula, retention time,Fragment ion accurate mass number, abundance of ions ratio;
(2) first class mode obtaining in step (1) is composed to data entirely, the fragment ion full scan mass spectrogram after examining is ledEnter in PCDL software, and be associated with corresponding agricultural chemicals information, build up TOF/MS accurate mass database, divide for data retrievalAnalyse;
(3) the full spectrum data of the mensuration testing sample Pesticides of sprouts of vegetable vegetables under the imposing a condition identical with step (1),Obtain chemical molecular formula, retention time, fragment ion accurate mass number, the abundance of ions ratio of its agricultural chemicals containing;
(4) to sample determination result through accurate mass database retrieval, the agricultural chemicals that detects that meets appraisal basis is carried out to standardReally qualification, thus the residual kind of agriculture in sprouts of vegetable vegetables obtained.
Preferably, by sprouts of vegetable vegetables being carried out to pre-treatment to obtain the mensuration testing sample of sprouts of vegetable vegetables, described inPre-treating method comprises following program:
Sample preparation: sprouts of vegetable vegetable sample extracts through the homogenate of 0.5-5 volume % acetate acetonitrile, through dehydration and centrifugal, denseAfter contracting, then through Carbon/NH2Column purification, acetonitrile+toluene wash-out remains of pesticide is made and is treated sample measuring liquid a after concentrating, filtering.
The beneficial effect that the present invention has:
1, the present invention taking the compound information such as high-resolution fragment ion accurate mass number, retention time, abundance of ions compare asCriterion of identification, 708 kinds of agricultural chemicals of foundation are detected and confirmation technical method, have thoroughly changed original taking compound reference material as referenceQualitative model, be that one does not need reference material contrast, fast, high flux, Detecting Pesticide new technology accurately and reliably.
2, the present invention is not needing, under the prerequisite of reference material contrast, to have realized qualitative detecting and the confirmation of 708 kinds of residues of pesticides,Can greatly save the cost of buying reference material, also environmental protection more, safety.
3, the present invention can complete 708 kinds of agricultural chemicals detectings, qualitative and confirmation in 1 hour, compare with traditional detection method,Can increase work efficiency hundreds of times.
4, the present invention can accomplish, to 708 kinds of disposable extraction and cleanings of residues of pesticides in sprouts of vegetable vegetable sample, once to enterSample detects, and compares with traditional detection method, can save testing cost, increase work efficiency hundreds of times.
5, the sprouts of vegetable vegetable sample Pesticides that the present invention can test comprises a class or a few class in following material: organicPhosphorus agricultural chemicals, carbamate chemicals for agriculture, benzimidazole agricultural chemicals, sulfonylurea pesticide, nicotinoids agricultural chemicals, cinerins agricultureThe agricultural chemicals of medicine and other types and metabolin.
6, the GC-QTOF/MS detection techniques index that the present invention sets up: sweep limits 50-600m/z; The accurate mass of measuringCan reach 0.0001m/z; Quality precision can be controlled in 20ppm; Sweep speed is 5 times per second in 50-600m/z scope.
7, a kind of application gas-chromatography level Four bar flight time mass spectrum (GC-Q-TOF/MS) rapid screening provided by the inventionIn sprouts of vegetable vegetables, the high resolution mass spectrum detection techniques of 708 kinds of residues of pesticides has obtained pushing away in 7 pesticide residue analysis laboratoriesWide and application, through the detecting checking to the 30 multiple commercially available sprouts of vegetable vegetable samples of Duo Ge provinces and cities, has obtained important residual detecingSurvey data.
8, simultaneously detecting 708 kinds of agricultural chemicals in, detecting sensitivity without exception lower than standard 10 micrograms/kilogram, meet various countries' agricultureThe requirement of the horizontal examination of the residual MRL of medicine.
Other features and advantages of the present invention are described in detail the detailed description of the invention part subsequently.
Brief description of the drawings
Accompanying drawing is to be used to provide a further understanding of the present invention, and forms a part for description, with tool belowBody embodiment one is used from explanation the present invention, but is not construed as limiting the invention. In the accompanying drawings:
Fig. 1 is the Shijiazhuang District commercially available mung bean sprouts sample GC-Q-TOF/MS the highest agricultural chemicals of technology frequency (chlopyrifos)Result example.
Fig. 2 is that the highest agricultural chemicals of the frequency that Shijiazhuang District commercially available mung bean sprouts sample GC-Q-TOF/MS technology detects (is poisoned with poisonTick) Information in Mass Spectra and the database result of comparing.
Detailed description of the invention
Below the specific embodiment of the present invention is elaborated. Should be understood that, described herein concreteEmbodiment only, for description and interpretation the present invention, is not limited to the present invention.
The invention provides the method for detecting that a kind of GC-Q-TOF/MS detects 708 kinds of residues of pesticides in sprouts of vegetable vegetables, itsBe characterised in that: set up 708 kinds of agricultural chemicals TOF/MS databases, simultaneously to 708 kinds of agricultural chemicals in sprouts of vegetable vegetables carry out qualitative detecting withConfirmation; The method comprises the following steps:
(1) standard sample of pesticide is mixed with respectively to the standard liquid of 1-5mg/L, imposes a condition lower to GC-QTOF/MS one-levelPattern is composed the full spectrum data of every kind of standard liquid of data determination entirely, obtain respectively every kind of agricultural chemicals chemical molecular formula, retention time,Fragment ion accurate mass number, abundance of ions ratio;
(2) first class mode obtaining in step (1) is composed to data entirely, the fragment ion full scan mass spectrogram after examining is ledEnter in PCDL software, and be associated with corresponding agricultural chemicals information, build up TOF/MS accurate mass database, divide for data retrievalAnalyse;
(3) the full spectrum data of the mensuration testing sample Pesticides of sprouts of vegetable vegetables under the imposing a condition identical with step (1),Obtain chemical molecular formula, retention time, fragment ion accurate mass number, the abundance of ions ratio of its agricultural chemicals containing;
(4) to sample determination result through accurate mass database retrieval, the agricultural chemicals that detects that meets appraisal basis is carried out to standardReally qualification, thus the residual kind of agriculture in sprouts of vegetable vegetables obtained.
Concrete, taking phenthoate dimephenthoate cidial (Phenthoate), as example illustrates, GC-QTOF/MS database of the present invention passes through followingMethod is set up:
1), at data acquisition interface, phenthoate dimephenthoate cidial (Phenthoate) reference material (1-5mg/L) is being specified to chromatogram mass spectrum barUnder part, carry out the collection of fragment ion full scan mass spectrogram through GC-QTOF/MS first class mode.
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-with 3-10 DEG C/min again280 DEG C, be then warming up to 110-150 DEG C with 20-50 DEG C/min, then be warming up to 200-280 DEG C with 3-10 DEG C/min, subsequently with 5-15 DEG C/min is warming up to 250-320 DEG C, keeps 3-10min, then is cooled to 200-290 DEG C with 5-20 DEG C/min, keeps 1-5min;Carrier gas: helium; Purity >=99.999%; 0.5-2mL/min; Injector temperature: 230-320 DEG C; Sample size 1 μ L; Input mode:Without split sampling, 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, then with 30 DEG C/minBe warming up to 130 DEG C, then be 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 with 10DEG C/min is cooled to 280 DEG C, keeps 2min; Carrier gas: helium, purity >=99.999%; Flow velocity: 1.2mL/min; Injection port temperatureDegree: 290 DEG C; Sample size 1 μ L; Input mode: without split sampling, solvent delay 6min.
Mass spectrum condition is: ion gun: EI (electron impact ionization source); Ion source temperature: 230-330 DEG C, electron bombardment ionization sourceEnergy: 50-100eV; Gas phase-mass spectrometer interface temperature: 200-320 DEG C; Scan mode: the mode of opening spectrogram with 2-10 per second is carried outTOF pattern full scan, mass number scope 50-600amu. Preferably, mass spectrum condition: ion gun: 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: with per second 5Open the mode of spectrogram and carry out TOF pattern full scan, mass number scope 50～600amu..
Described fragment ion full scan mass spectrogram collection is the mass spectrogram information of phenthoate dimephenthoate cidial (Phenthoate) reference material,Comprise compound chemistry molecular formula, retention time, fragment ion accurate mass number, abundance of ions ratio.
2) in the qualitative software of AgilentMassHunter (instrument carries), invocation target agricultural chemicals (Phenthoate) is adoptedCollection file, and definite target agricultural chemicals chromatographic peak.
3) open after target agricultural chemicals mass spectrogram background correction, use the low resolution spectrum storehouse of NIST11 (instrument carries) to search forTo obtain the information of compound, and verify library searching to compound whether conform to actual.
4) verify mass spectrogram, target compound mass spectrogram can show the fragment ion matching through the low resolution spectrum of NIST11 storehouseMolecular formula and accurate mass number, carry out necessary amendment by " EditPeakAnnotion " window to ion information, such asAccurate mass is counted the excessive ion of deviation, may mean that the automatic Formula structure of calculating of software is wrong, now can manually existIn " Formula " row, input correct information, or " IronType " changed to Unknown.
5) the fragment ion full scan mass spectrogram after examining imports in PCDL software (instrument carries), and believes with phenthoate dimephenthoate cidialManner of breathing association, builds up TOF/MS database. The mass spectrogram information of 708 kinds of agricultural chemicals in database, for details see attached table 1.
Preferably, described agricultural chemicals is 708 kinds of agricultural chemicals shown in table 1. The method that forms these agricultural chemicals can be according to as aboveThe mode that forms the TOF/MS database of phenthoate dimephenthoate cidial (Phenthoate) is carried out.
According to the present invention, the mass spectrogram information exchange of obtained reference material is crossed to PCDL software and mutually close with corresponding agricultural chemicalsThe method of connection can be carried out according to PCDL software document, and the present invention does not repeat them here.
According to the present invention, preferred, the group that described sprouts of vegetable vegetables select free horse mung bean sprouts, Chinese toon bud and toothed burclover to formIn.
Table 1
According to the present invention, with set up identical imposing a condition under the full spectrums of mensuration testing sample Pesticides of sprouts of vegetable vegetablesData, obtain chemical molecular formula, retention time, fragment ion accurate mass number, the abundance of ions ratio of its agricultural chemicals containing; CallThe TOF/MS database of having set up, according to TOF/MS database, to target compound retrieval by header, search argument retention time is inclined to one sidePoor be defined as ± 0.15min, be defined as ± 20ppm of accurate mass deviation, software can be according to every kind of compound retention time, accurateThe measured value of the key elements such as quality automatically provides each retrieval that detects ion with the deviation of theoretical value in TOF/MS database and matesObtain score value. Detect ion score mean value and be this compound integrate score, specify that each compound obtains score value > 60 and at leastThree ions detect, i.e. positive detecting is accurate according to the method for the qualitative detecting Residual Pesticides in Farm Produce of TOF/MS databaseDegree, reaches more than 90%.
According to the present invention, by sprouts of vegetable vegetables being carried out to pre-treatment to obtain the mensuration testing sample of sprouts of vegetable vegetables,Described pre-treating method preferably includes following program:
Acetonitrile solution by sprouts of vegetable vegetable sample through containing 0.5-5 volume % acetic acid (is called for short 0.5-5 volume % acetic acid secondNitrile) homogenate extracts, through dehydration and centrifugal, concentrate after, then through Carbon/NH2Column purification, acetonitrile+toluene wash-out remains of pesticide, warpAfter concentrating again, filtering, make and treat sample measuring liquid;
Wherein, preferred, described sprouts of vegetable vegetable sample amount is 5-20g, the use of the acetate acetonitrile of described 0.5-5 volume %Amount is that the consumption of 30-50mL is preferably 35-45ml; And the fruits and vegetables sample that adds acetate acetonitrile is placed in to centrifuge tube carries out homogenateProcess. Described homogenate extraction conditions is preferably: rotating speed 12000-15000rpm, time 0.5-2min.
Wherein, described through dewatering preferably by add the sodium chloride of 0.5-5g, the nothing of 2-8g in acetate acetonitrile extractWater magnesium sulfate, and the 4-6min that vibrates realizes. Then by the dehydration sample obtaining centrifugal 1-10min under 4000-4500rpm, obtainTo supernatant, and the supernatant 10-30mL obtaining rotary evaporation in the water-bath of 20-60 DEG C is concentrated into 0.5-2mL.
Wherein, described Carbon/NH2Column purification step is preferably as follows: at Carbon/NH2In post, add about 1-3cm preferredThe high anhydrous sodium sulfate of 2cm. First with the preferred 4mL acetonitrile+toluene of 2-6mL (preferably 2.5-3.5: 1, more preferably 3: 1, v/v) drip washingSPE post, and discard efflux, in the time that liquid level arrives the top of sodium sulphate, rapidly sample concentration liquid is transferred on decontaminating column, underConnecing new heart bottle graft receives. Wash with the preferred 2mL acetonitrile+toluene of 2-6mL (preferably 2.5-3.5: 1, more preferably 3: 1, v/v) more at every turnWash sample liquid bottle three times, and cleaning solution is moved in SPE post. Again with the preferred 25mL acetonitrile+toluene of 20-30mL (preferably 2.5-3.5:1, more preferably 3: 1, v/v) wash-out agricultural chemicals, merge eluent in heart bottle.
Wherein, described in treat that sample measuring liquid is further preferably prepared in the following way, by as above obtain eluent existIn 30-50 DEG C of water-bath, rotation is concentrated into about 0.5-2mL, and concentrate is placed under nitrogen and is dried up, and adds acetonitrile+0-of 1-5mL5% formic acid water (2+ (7-9), preferably 2+8, v/v) mixes, and through preferably constant volume after 0.2 μ m membrane filtration of 0.18-0.22 μ m, obtainsTreat sample measuring liquid.
Below will describe the specific embodiment of the present invention in detail.
Embodiment 1
The present embodiment is for illustrating 708 kinds of residual detecing of agriculture of GC-Q-TOF/MS examination sprouts of vegetable vegetables provided by the inventionSurvey method
(1) take 10g Shijiazhuang District mung bean sprouts sample (being accurate to 0.01g), in 80ml centrifuge tube, add 40mLThe acetonitrile solution of 1% acetic acid, with high-speed homogenization machine 13500r/min, 1min is extracted in homogenate, adds 1g sodium chloride, 4g anhydrous slufuric acidMagnesium, vibration 5min, centrifugal 5min under 4200r/min, gets supernatant 20mL, and in 40 DEG C of water-baths, rotary evaporation is concentrated into approximately1mL, to be clean.
(2) at Carbon/NH2In post, add the high anhydrous sodium sulfate of about 2cm. First use 4mL acetonitrile+toluene (3+1, v/v) to drenchWash SPE post, and discard efflux, in the time that liquid level arrives the top of sodium sulphate, rapidly sample concentration liquid is transferred in SPE post,Under connect new heart bottle graft and receive. Use again 2mL acetonitrile+toluene (3+1, v/v) washing sample liquid bottle three times at every turn, and cleaning solution is moved intoIn SPE post. On post, connect 50mL liquid reservoir, with 25mL acetonitrile+toluene (3+1, v/v) wash-out agricultural chemicals and related chemicals, closeAnd in heart bottle, and rotation is concentrated into about 0.5mL in 40 DEG C of water-baths.
(3) concentrate is placed under nitrogen and dries up, and adds acetonitrile+water (2+8, v/v) of 2mL to mix, through 0.2 μ m membrane filtrationRear constant volume, obtains testing sample solution.
(4) GC-Q-TOF/MS operating condition, separates and obtains the credit of the each fragment ion of each agricultural chemicals in mung bean sprouts sampleMinor, retention time, fragment ion accurate mass number, abundance of ions 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 5 DEG C/min250 DEG C, be warming up to 300 DEG C with 10 DEG C/min subsequently, keep 5min, then be cooled to 280 DEG C with 10 DEG C/min, keep 2min; CarryGas: helium, purity >=99.999%; Flow velocity: 1.2mL/min; Injector temperature: 290 DEG C; Sample size 1 μ L; Input mode: nothingSplit sampling, solvent delay 6min.
Mass spectrum condition: mass spectrum condition: ion gun: EI (electron impact ionization source); Ion source temperature: 280 DEG C, electronics bombardmentSource energy: 70eV; Gas phase-mass spectrometer interface temperature: 290 DEG C; Scan mode: carry out TOF pattern in the mode of 5 spectrograms per second completeScanning, mass number scope 50～600amu..
(5) according to the database of table 1 to acquired information retrieval by header in step (4), search argument retention time deviation limitBe decided to be ± 0.5min, be defined as ± 100ppm of accurate mass deviation, detects at least three fragment ions, and according to every kind of compoundThe deviation of theoretical value in the measured value of each key element and table 1 database, provides retrieval matching score value automatically, retrieval matching score valueThe compound of > 60, confirms to detect this target compound. Testing result is in table 2.
Taking the higher chlopyrifos of frequency as example, as shown in Figure 1, in mung bean sprouts, detect 7 fragment ion information, useDatabase retrieval, compares with chlopyrifos, and 7 fragment ion molecular weight deviation scores are all more than 99 points, and retention time deviation existsIn 0min, the fragment ion mass deviation that abundance is the highest is-1.28ppm (as shown in Figure 2) to confirm having in mung bean sproutsChlopyrifos detects.
Embodiment 2
The present embodiment is for illustrating 708 kinds of residual detecing of agriculture of GC-Q-TOF/MS examination sprouts of vegetable vegetables provided by the inventionSurvey method
To residual detecting of agriculture with in batch mung bean sprouts sample, different according to the method in embodiment 1, pre-treatment stepRapid as follows:
(1) accurately take 10.00g sample in centrifuge tube, add 50mL water, vibrate until sample is fully infiltrated by water.After freezing 10min, pipette 50mL acetonitrile to centrifuge tube, then centrifuge tube is placed on vortex mixing shaker, with 2000rpmRate oscillation 1min. Centrifuge tube is placed under-10 DEG C of conditions and keeps 10min, then in centrifuge tube, add 20g anhydrous magnesium sulfateWith 5g sodium chloride, 5g natrium citricum and 2.5g Sodium citrate, immediately on whirlpool mixing shaker, with 2000rpm speedVibration 2min, then, with the centrifugal 10min of 4000rpm speed, obtains supernatant.
(2) pipette supernatant 1.0mL in 1.5mL centrifuge tube, add the PSA absorption of 150mg anhydrous magnesium sulfate and 25mgAgent, on whirlpool mixing shaker with 2000rpm rate oscillation 2min, with the centrifugal 2min of 6000rpm speed. Draw supernatant warp0.22 μ m organic phase membrane filtration, pipettes 200 μ L, adds 100 μ L acetonitriles, is diluted to 1.0mL with ultra-pure water, obtains testing sampleSolution.
Testing result is in table 2.
Table 2
Can be found out by above data, adopt method of the present invention, especially preferred method can more effectively detectGo out the residual kind of agricultural product middle peasant, and there is lower detectability.
More than describe the preferred embodiment of the present invention in detail, still, the present invention is not limited in above-mentioned embodimentDetail, within the scope of technical conceive of the present invention, can carry out multiple simple variant to technical scheme of the present invention, thisA little simple variant all belong to protection scope of the present invention.
It should be noted that in addition, each the concrete technical characterictic described in above-mentioned detailed description of the invention, at lance notIn the situation of shield, can combine by any suitable mode. For fear of unnecessary repetition, the present invention to various canThe combination explanation no longer separately of energy.
In addition, between various embodiment of the present invention, also can be combined, as long as it is without prejudice to thisThe thought of invention, it should be considered as content disclosed in this invention equally.
Claims (10)
1. GC-Q-TOF/MS detects a method for detecting for 708 kinds of residues of pesticides in sprouts of vegetable vegetables, it is characterized in that: set up708 kinds of agricultural chemicals TOF/MS databases carry out qualitative detecting and confirmation to 708 kinds of agricultural chemicals in sprouts of vegetable vegetables simultaneously; The method bagDraw together following steps:
(1) standard sample of pesticide is mixed with respectively to the standard liquid of 1-5mg/L, imposes a condition lower to GC-QTOF/MS first class modeEntirely compose the full spectrum data of every kind of standard liquid of data determination, obtain respectively chemical molecular formula, retention time, the fragment of every kind of agricultural chemicalsExact mass of ion number, abundance of ions ratio;
(2) first class mode obtaining in step (1) is composed to data entirely, the fragment ion full scan mass spectrogram after examining importsIn PCDL software, and be associated with corresponding agricultural chemicals information, build up TOF/MS accurate mass database, divide for data retrievalAnalyse;
(3) the full spectrum data of the mensuration testing sample Pesticides of sprouts of vegetable vegetables under the imposing a condition identical with step (1), obtainThe chemical molecular formula of its agricultural chemicals containing, retention time, fragment ion accurate mass number, abundance of ions ratio;
(4) to sample determination result through accurate mass database retrieval, the agricultural chemicals that detects that meets appraisal basis is accurately reflectedFixed, thus the residual kind of agriculture in sprouts of vegetable vegetables obtained.
2. method according to claim 1, wherein, described agricultural chemicals is 708 kinds of agricultural chemicals shown in table 1.
3. method according to claim 1, wherein, described sprouts of vegetable vegetables select free mung bean sprouts, Chinese toon bud and toothed burclover instituteIn the group of composition.
4. method according to claim 1, wherein, measures the condition bag of the full spectrum data of every kind of standard liquid PesticidesDraw together:
(1) chromatographic condition: chromatographic column: VF-1701MS; Chromatographic column heating schedule: 30-50 DEG C keeps 0.5-5min, then with 20-50 DEG C/min is warming up to 110-150 DEG C, then is warming up to 200-280 DEG C with 3-10 DEG C/min, is warming up to subsequently with 5-15 DEG C/min250-320 DEG C, keeps 3-10min, then is cooled to 200-290 DEG C with 5-20 DEG C/min, keeps 1-5min; Carrier gas: helium; StreamSpeed: 0.5-2mL/min; Injector temperature: 230-320 DEG C: without split sampling, solvent delay 2-10min;
(2) mass spectrum condition: ion gun: 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: the mode of opening spectrogram with 2-10 per second is carried out TOF pattern full scan, qualityNumber scope 50-600amu.
5. method according to claim 1, wherein, TOF/MS accurate mass database can will be measured from testing sampleThe measured value of the retention time of the fragment ion arriving, accurate mass number, abundance of ions ratio, in TOF/MS accurate mass databaseThe corresponding key element theoretical value retrieval comparison of fragment ion, provides the score of each fragment ion, and then to provide average mark be targetThe matching degree of compound obtains score value; Wherein, the retention time deviation setting of fragment ion is at ± 0.5min, and accurate mass is counted deviationBe set as ± 100ppm.
6. method according to claim 5, wherein: by the following method the agricultural chemicals that detects that meets appraisal basis is carried out to standardReally qualification:
By characteristic ion being detected to situation and degree of conformity, provide score value, agricultural chemicals is carried out to qualitative identification, its method programFor: for the detecting of target compound in sample, be that retention time, the abundance of compound that flight time mass spectrum is recorded is the highest7 fragment ion accurate mass numbers, the result of fragment ion abundance ratio, compare with TOF/MS database retrieval, result is inspectionGo out at least three fragment ions, and integrate score is greater than 60 agricultural chemicals and is confirmed as detecting agricultural chemicals.
7. method according to claim 1, by carrying out pre-treatment to obtain the survey of sprouts of vegetable vegetables to sprouts of vegetable vegetablesDetermine testing sample, described pre-treating method comprises following program:
Sample preparation: sprouts of vegetable vegetable sample through the homogenate of 0.5-5 volume % acetate acetonitrile extract, through dehydration and centrifugal, concentrate after,Again through Carbon/NH2Column purification, acetonitrile+toluene wash-out remains of pesticide is made and is treated sample measuring liquid a after concentrating, filtering.
8. method according to claim 7, wherein: described sprouts of vegetable vegetable sample amount 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, describedly refer in 0.5-5 volume % acetate acetonitrile extract and add 0.5-5g sodium chloride through dehydration, 2-8g withoutWater magnesium sulfate, vibration 1-10min;
Preferably, the described centrifugal centrifugal 1-10min under 3000-5000r/min that refers to;
Preferably, described concentrated referring to got centrifuged supernatant 10-30mL, and in 20-60 DEG C of water-bath, rotary evaporation is concentrated into approximately0.5-2mL。
9. method according to claim 8, wherein: described Carbon/NH2Column purification step is as follows: at Carbon/NH2PostIn add the high anhydrous sodium sulfate of about 1-3cm, first use a certain proportion of acetonitrile of 2-6mL and toluene mixed solution drip washing SPE post, and abandonRemove efflux, in the time that liquid level arrives the top of sodium sulphate, rapidly sample concentration liquid be transferred on decontaminating column, under connect container and connectReceive; Wash sample liquid bottle three times with 2-6mL acetonitrile and toluene mixed solution more at every turn, and cleaning solution is moved in SPE post. Use again excellentSelect certain volume acetonitrile and the toluene mixed solution wash-out agricultural chemicals of 20-30ml, merge eluent in container;
Preferably, described a certain proportion of acetonitrile and toluene mixed solution, acetonitrile: volume of toluene is than being 2.5-3.5: 1.
10. method according to claim 8, wherein: described through more concentrated, make after filtering and treat that sample measuring liquid a refers to rightRequire 9 eluents that obtain, in 30-50 DEG C of water-bath, rotation is concentrated into about 0.5-2mL, and concentrate is placed in to nitrogen blowing downDry, add a certain proportion of acetonitrile of 1-5mL and 0-5 volume % formic acid water mixed solution to mix, through 0.18-0.22 μ m filter membrane mistakeConstant volume after filter, 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 water2∶7-9。
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