CN105675738A - LC-Q-TOF/MS detection technology for 544 pesticide residues in stem vegetables - Google Patents
LC-Q-TOF/MS detection technology for 544 pesticide residues in stem vegetables Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
Abstract
The invention provides an LC-Q-TOF/MS detection technology for 544 pesticide residues in stem vegetables. The technology includes the steps of: 1) under a TOF/MS mode in the LC-Q-TOF/MS, respectively measuring retention time of each pesticide reference substance under the certain chromatography-mass spectrometry conditions; 2) determining the ionization formation and chemical formula of the compound with an ESI source to obtain accurate mass number of a parent ion of each compound to form a TOF/MS database; 3) under a Q-TOF/MS mode, respectively collecting fragment ion mass spectrograms under 3-5 different collision energy of each pesticide reference substance, and introducing the collected information to PCDL software to obtain a Q-TOF/MS database; 4)comparing the retention time, primary mass spectrum and secondary mass spectrum of the sample of the stem vegetables to determine whether the pesticide residues exist in the sample or not, and meanwhile, performing secondary determination to the compound having higher primary score, and if secondary score is high, determining existence of the pesticide residues. The technology is quick, high-throughput, high-precision and high-reliability, and can be used for accurately screening the pesticides in the stem vegetables.
Description
Technical field
The present invention relates to food hygiene, agricultural product security detection field, particularly to a kind of for the method for detecting of 544 kinds of pesticide residues in stem class vegetable.
Background technology
Global environment detection system/food item (GlobalEnvironmentMonitoringSystem is jointly established as far back as World Health Organization (WHO) (WHO) in 1976, food and agricultural organization (FAO) and United Nations Environment Programme (UNEP); GEMS/Food); it is intended to grasp member state's food pollution situation; understand food contaminant intake; protection health, give a impetus to 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 residues presenting kind and gets more and more, increasingly stricter development trend of limiting the quantity, the Pesticide Residue threshold that namely international trade is set up is more and more higher. Present European Union has formulated 162248 MRL standards of 839 kinds of pesticide, and the U.S. has formulated 39147 MRL standards of 351 kinds of pesticide, and Japan has formulated 51600 multinomial MRL standards of 579 kinds of pesticide, and China has issued 3650 MRL standards of 381 kinds of pesticide for 2014. At present, commonly used in the world uniform limit limitation is 10 μ g/kg. Therefore, the quick the Detection Technologies of Pesticide Residues of high flux 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 is to realize the optimized analysis means that the many residuals of high flux quickly detect.
Current pesticide residue analysis is many based on gas chromatogram, liquid chromatograph, gas chromatography-mass spectrum and LC-MS-MS. These detection techniques all carry out qualitative firstly the need of standard sample of pesticide comparison. Such as, it is accomplished by preparing corresponding 100 kinds of standard sample of pesticide comparison to the detection of 100 kinds of pesticide, and the pesticide outside these 100 kinds all can be missed. In the real work of agricultural and veterinary chemicals results from residue tests room, most laboratorys are all without laying in hundreds of standard sample of pesticide, and its reason is that standard sample of pesticide is not only expensive, and effect duration only has 2,3 years, it is necessary to overlapping investment.The standing standard sample of pesticide of common laboratory only has tens kinds, and the pesticide species of its daily monitoring is also just only limited to these tens kinds, thereby results in food safety monitoring leak.
The present inventor team concentrates on studies through for many years, have developed the residual detection techniques of 544 kinds of pesticide in the accurate mass data base of two kinds of 544 kinds of pesticide and a kind of stem class vegetable, achieve do not need standard control, can to more than 500 kinds pesticide residues in stem class vegetable, rapid detection detection, meets the urgent need that Agricultural Products Pesticide Residues high flux quickly detects simultaneously.
Summary of the invention
The present invention, have rated on the basis of mass spectrum behavior characteristics of conventional 1000 Multiple Pesticides in the world, establishes 544 kinds of pesticide TOF/MS and Q-TOF/MS accurate mass data base innovatively; Establish with compound Information in Mass Spectras such as high-resolution accurate mass number, Isotopic Distribution and isotope abundances for criterion of identification innovatively, according to the LC-Q-TOF/MS technical method that 544 kinds of pesticide are detected and confirmed by TOF/MS and Q-TOF/MS accurate mass data base; Develop matched stem class Pesticide Residues in Vegetables high flux sample preparation technology innovatively. Achieve and have only to a sample preparation, single injected sampling detection, just can to the rapid detection of 500 Multiple Pesticides in stem class vegetable, and the pesticide detected can be carried out Q-TOF/MS data base's confirmation, revolutionize original qualitative model being reference with compound standard thing, it is that a kind of reference material that do not need compares, quick, high flux, accurately and reliably Detecting Pesticide new technique.
One, novelty of the present invention is in that:
1, establish 544 kinds of pesticide accurate masses innovatively and can reach the accurate mass data base of 0.0001m/z, including TOF/MS data base and Q-TOF/MS data base.
2, TOF/MS data base comprises the full scan mass spectrum information of the English name of 544 kinds of pesticide, chemical molecular formula, retention time, accurate mass number, Isotopic Distribution, isotope abundance and parent ion and fragment ion, contains much information, accurately comprehensively.
3, the full scan mass spectrum information of every kind of pesticide fragment ion in Q-TOF/MS data base, is by full scan mass spectrum under eight different collision energies is preferred, fragment ion full scan figure under informative four the different collision energies of ion obtained.
4, establish innovatively with compound Information in Mass Spectras such as high-resolution accurate mass number, Isotopic Distribution and isotope abundances for criterion of identification, according to the LC-Q-TOF/MS technical method that 544 kinds of pesticide are detected and confirmed by TOF/MS and Q-TOF/MS accurate mass data base, revolutionize original qualitative model being reference with compound standard thing, it is that a kind of reference material that do not need compares, quick, high flux, accurately and reliably Detecting Pesticide new technique.
5, the residual detection techniques method that the present invention sets up can according to information such as the retention time of target compound, accurate mass number, Isotopic Distribution and isotope abundances, by retrieving comparison with the corresponding informance of compound in TOF/MS data base, provide the matching degree score value of target compound. Score value according to target compound, it is achieved the qualitative detecting to pesticide.
6, the residual detection techniques method that the present invention sets up can according to the full scan mass spectrum information of target compound fragment ion, by with the full scan mass spectrum information retrieval comparison of compound fragment ion in Q-TOF/MS data base, provide the matching degree score value of target compound, it is achieved the qualitative confirmation to target compound.Compare the qualitative confirmation method of traditional detection, owing to have employed full scan mass spectrum information comparison, there is higher qualitative accuracy.
7, the Residual Pesticides in Farm Produce high flux sample preparation technology supporting with remaining detection techniques method is developed. Achieve and have only to a sample preparation, twice sample detection, it is possible to the rapid detection of 500 Multiple Pesticides in agricultural product and confirmation.
8, the main detection techniques index of LC-Q-TOF/MS: sweep limits 50-1600m/z; The accurate mass measured is up to 0.0001m/z; Within Mass accuracy can be controlled in the 10ppm of molecular mass; Scanning speed ranges for 4 times per second at 50-1600m/z. The main detection techniques feature of LC-Q-TOF/MS: sample is once prepared, can complete the detecting of 544 kinds of pesticide, qualitative with confirmation in 1 hour; More than 80% pesticide confirmation point reaches more than 10; In the 544 kinds of pesticide detected at the same time, 327 kinds of pesticide detecting sensitivity, lower than the micro-g kg of uniform limit 10, accounting 64.1%, substantially meet the requirement of the horizontal examination of various countries pesticide residues MRL.
Two, the technical solution used in the present invention:
1, accurate mass data base is set up
The foundation of 1.1TOF/MS data base
Under TOF/MS pattern, measure every kind of pesticide standard thing retention time under specifying chromatograph Mass Spectrometry Conditions respectively, it is determined that the ionized form (+H ,+NH under this compound ESI source4,+Na) and chemical formula, obtaining the accurate mass number of every kind of compound parent ion, isotopic peak is distributed and abundance ratio. The information such as the title of 544 kinds of pesticide, retention time, molecular formula, accurate mass number, isotopic peak distribution and abundance ratio are imported in data base's CSV software document, forms TOF/MS data base.
The foundation of 1.2Q-TOF/MS data base
Sequentially input the parent ion of 544 kinds of pesticide in TOF/MS data base at AgilentMassHunter data acquisition interface, under Q-TOF/MS pattern, to 544 kinds of pesticide respectively under 8 collision energies, carry out fragment ion full scan mass spectrum collection. Under informative 3-5 the different collision energies of ion, fragment ion full scan mass spectrum imports in database software file preferably wherein, and is associated with corresponding pesticide information, builds up Q-TOF/MS data base.
2, setting up with compound information such as high-resolution accurate mass number, Isotopic Distribution and isotope abundances for criterion of identification, 544 kinds of pesticide LC-Q-TOF/MS detect and confirmation technical method, and technical step includes:
2.1 sample preparations: stem class vegetable sample extracts through 1% acetate acetonitrile, Carbon/NH2Column purification, acetonitrile+toluene (3+1, v/v) eluting residual pesticide, make after concentrated, dissolving and filtration and treat sample measuring liquid;
Described stem class vegetable includes Germinatus Phragmitis;
2.2 chromatographic mass spectrometry detections: adopt the liquid chromatograph being furnished with reversed phase chromatographic column (ZORBAXSB-C18,2.1mm × 100mm, 3.5 μm) to separate the pesticide treating in sample measuring liquid, are furnished with the LC-Q-TOF-MS detection of electric spray ion source;
The qualitative method for detecting of 2.3TOF/MS data base: call the TOF/MS data base set up in the qualitative software of AgilentMassHunter, according to TOF/MS data base to target compound retrieval by header, search argument retention time deviation is defined to ± 0.5min, accurate mass deviation is defined to ± 10ppm, ionized form selection+H ,+NH4+ Na pattern, software can according to the measured value of the key elements such as every kind of Compound Retention time, accurate mass and the deviation of theoretical value in TOF/MS data base, and Isotopic Distribution and four parameters of isotope ratio, by scientifically setting each key element weight (wherein the weight of accurate mass number deviation is the highest) in matching degree score calculates, provide retrieval matching score value, the compound that retrieval matching score value is higher, for doubtful detection compound, according to the method accuracy of the qualitative detecting Residual Pesticides in Farm Produce of TOF/MS data base, reach more than 90%;
The qualitative confirmation method of 2.4Q-TOF/MS data base: under TargetedMS/MS drainage pattern, input the parent ion of doubtful detection compound, retention time and best collision energy, treat sample measuring liquid again to detect, by detection mass spectrum and the full scan mass spectrum of compound fragment ion in Q-TOF/MS data base, mate when mirror image contrast, whether consistent with spectrum storehouse according to characteristic ion distribution, the ratio of secondary fragment accurate mass number deviation and fragment ion provides score, its matching score value is higher, namely confirms to detect this target compound. This confirmation method adopts full scan mass spectrum coupling confirmation, makes more than 80% pesticide confirmation point can reach more than 10, and ratio 4 confirmation point requirements of standard have higher qualitative accuracy.
Three, the technology contents of the present invention:
The present invention provides the accurate mass data base of two kinds of 544 kinds of pesticide and one for detecting 544 kinds of pesticide residues detection techniques in stem class vegetable.
1, two kinds of accurate mass data bases provided by the invention are TOF/MS data base and Q-TOF/MS data base.
1.1TOF/MS database technology content includes: the chemical formula of 544 kinds of pesticide, accurate mass number, the information such as retention time and pesticide English name. In TOF/MS data base, 3 kinds of compounding techniques content example are as follows:
1.2Q-TOF/MS database technology content includes: 544 kinds of pesticide fragment ion information mass spectrum under 4 collision energies. Q-TOF/MS database technology content example is as shown in Figure 1.
2, the invention provides 544 kinds of pesticide LC-Q-TOF/MS detectings and confirmation technical method in a kind of stem class vegetable. Detecting and confirmation technical method content include as follows:
2.1 sample preparations
2.1.1 stem class vegetable sample takes edible part chopping, mixing, seals, indicates labelling.
2.1.2 10g stem class vegetable sample (being accurate to 0.01g) is weighed, in 80mL centrifuge tube, add 40mL1% acetate acetonitrile, with high-speed homogenization machine 13500r/min, 1min is extracted in homogenate, adding 1g sodium chloride, 4g anhydrous magnesium sulfate, vibrate 5min, centrifugal 5min under 4200r/min, taking supernatant 20mL, in 40 DEG C of water-baths, rotary evaporation is concentrated into about 1mL, to be clean.
2.1.3 in Carbon/NH2 post, add about 2cm height anhydrous sodium sulfate. First with 4mL acetonitrile+toluene (3+1, v/v) drip washing SPE post, and discard effluent, when liquid level arrives the top of sodium sulfate, rapidly sample concentration liquid be transferred on decontaminating column, under connect new Cor Gigeriae Galli bottle graft and receive. Wash sample liquid bottle three times with 2mL acetonitrile+toluene (3+1, v/v), and cleaning mixture is moved in SPE post every time again. Post connects 50mL liquid reservoir, then with 25mL acetonitrile+toluene (3+1, v/v) eluting pesticide, is incorporated in Cor Gigeriae Galli bottle, and in 40 DEG C of water-baths spin concentration extremely about 0.5mL.
2.1.4 concentrated solution is placed under nitrogen and dries up, add acetonitrile+0.1% formic acid water (2+8, the v/v) mixing of 2mL, constant volume after 0.2 μm of membrane filtration, obtain testing sample solution a.
2.2LC-Q-TOF/MS detects
2.2.1 liquid chromatograph separates: testing sample solution a separates by being furnished with the liquid chromatographic system of reversed phase chromatographic column (ZORBAXSB-C18,2.1mm × 100mm, 3.5 μm); Mobile phase A is ammonium acetate-0.1% formic acid-water of 5mM; Mobile phase B is acetonitrile; Liquid chromatograph gradient elution program is: 0min:1%B, 3min:30%B, 6min:40%B, 9min:40%B, 15min:60%B, 19min:90%B, 23min:90%B, 23.01min:1%B, runs 4min afterwards;Flow velocity is 0.4mL/min; Column temperature: 40 DEG C; Sample size: 10 μ L.
2.2.2 Mass Spectrometer Method: Agilent6530LC-Q-TOF/MS is at electron spray ionisation positive ion mode (ESI+); Capillary voltage: 4000V; Dry temperature: 325 DEG C; Dry gas stream amount 10L/min, sheath gas flow velocity 11L/min, sheath gas temperature is 325 DEG C; Atomization gas pressure 40psi, taper hole voltage 60V, fragmentation voltage 140V. Full scan karyoplasmic ratio ranges for 50-1600m/z, and adopts interior mark reference solution that instrument quality precision is carried out real time correction. By AgilentMassHunterWorkstationSoftware (versionB.05.00) to mass spectrometric data collection and process.
The qualitative detecting of 2.3TOF/MS data base
Calling the TOF/MS data base set up in the qualitative software of AgilentMassHunter, arrange search argument retention time deviation for ± 0.5min, accurate mass deviation is ± 10ppm, ionized form selection+H ,+NH4,+Na pattern; According to TOF/MS data base, the sample solution mass spectrometric data of above-mentioned collection being carried out retrieval by header, go out every kind of compound retrieval matching score value by computed in software, the compound of retrieval matching score value > 70, for doubtful detection compound.
The qualitative confirmation of 2.4Q-TOF/MS data base
Under TargetedMS/MS drainage pattern, input the parent ion of doubtful detection compound, retention time and best collision energy, testing sample solution a is detected again, by detection mass spectrum and the full scan mass spectrum of compound fragment ion in Q-TOF/MS data base, the coupling confirmation when mirror image contrast. Its matching score value > 70, namely confirms to detect this target compound.
The present invention is had the beneficial effect that compared to existing technology
1, the present invention is with compound information such as high-resolution accurate mass number, Isotopic Distribution and isotope abundances for criterion of identification, the 500 Multiple Pesticides detectings set up and confirmation technical method, revolutionize original qualitative model being reference with compound standard thing, it is that a kind of reference material that do not need compares, quick, high flux, accurately and reliably Detecting Pesticide new technique;
2, the present invention is under the premise not needing reference material to compare, it is achieved that the qualitative detecting of 500 Multiple Pesticides residuals and confirmation, it is possible to be greatly saved the cost buying reference material, also more environmentally-friendly, safety;
3, the present invention can complete 500 Multiple Pesticides detectings in 1 hour, qualitative with confirmation, compared with traditional detection method, it is possible to improve work efficiency hundreds times;
4, the present invention can accomplish 500 Multiple Pesticides in stem class vegetable sample are remained disposable extraction and cleaning, and single injected sampling detects, compared with traditional detection method, it is possible to saves testing cost, improve work efficiency hundreds times;
5, in 544 kinds of pesticide that the present invention detects simultaneously, there are 327 kinds of pesticide detecting sensitivity lower than 10 micro-g kg, accounting 64.1%, substantially meet the requirement of the horizontal examination of various countries pesticide residues MRL;
6, the present invention is under the premise not needing pesticide standard thing to compare, according to accurate mass data base to the qualitative of target compound and confirmation, making more than 80% pesticide confirmation point reach more than 10, whole pesticide meet European Union's requirement to compound 4 confirmation point, and qualitative results is more accurately and reliably;
7, the stem class vegetable sample Pesticides that the present invention can test includes the class in following material or a few class: organophosphorus insecticide, carbamate chemicals for agriculture, Ion pairing, sulfonylurea pesticide, nicotinoids pesticide, cinerins pesticide and other kinds of pesticide and metabolite;
8, the detection techniques index that the present invention sets up: sweep limits 50-1600m/z;The accurate mass measured is up to 0.0001m/z; Within Mass accuracy can be controlled in the 10ppm of molecular mass; Scanning speed ranges for 4 times per second at 50-600m/z;
9, the accurate mass data base of two kinds of 544 kinds of pesticide provided by the invention and one are used for detecting in stem class vegetable 544 kinds of pesticide residues detection techniques and are promoted at 7 pesticide residue analysis laboratorys and apply, through the detecting checking to 30 more than 500 commercially available stem class vegetable samples of provinces and cities, achieve important residual detecting data.
Accompanying drawing explanation
Fig. 1 Germinatus Phragmitis sample full scan pattern TOF shot chart;
Two grades of acquisition method exemplary plot of doubtful pesticide in Fig. 2 Germinatus Phragmitis sample;
Fig. 3 Germinatus Phragmitis sample detects two grades of Q-TOF/MS of pesticide and confirms shot chart.
Detailed description of the invention
Embodiment 1
In stem class vegetable (for Germinatus Phragmitis), 544 kinds of pesticide (see table 1) LC-Q-TOF/MS detectings and confirmation technology implementation example, comprise the steps:
1, the concrete steps of Sample Pretreatment Technique:
1.1 Germinatus Phragmitis samples take edible part chopping, mixing, seal, indicate labelling;
1.2 weigh 10g Germinatus Phragmitis sample (being accurate to 0.01g), in 80mL centrifuge tube, add 40mL1% acetate acetonitrile, with high-speed homogenization machine 13500r/min, 1min is extracted in homogenate, adding 1g sodium chloride, 4g anhydrous magnesium sulfate, vibrate 5min, centrifugal 5min under 4200r/min, taking supernatant 20mL, in 40 DEG C of water-baths, rotary evaporation is concentrated into about 1mL, to be clean.
1.3 at Carbon/NH2Post adds about 2cm height anhydrous sodium sulfate. First with 4mL acetonitrile+toluene (3+1, v/v) drip washing SPE post, and discard effluent, when liquid level arrives the top of sodium sulfate, rapidly sample concentration liquid be transferred in SPE post, under connect new Cor Gigeriae Galli bottle graft and receive. Wash sample liquid bottle three times with 2mL acetonitrile+toluene (3+1, v/v), and cleaning mixture is moved in SPE post every time again. Post connects 50mL liquid reservoir, with 25mL acetonitrile+toluene (3+1, v/v) eluting pesticide and related chemicals, is incorporated in Cor Gigeriae Galli bottle, and in 40 DEG C of water-baths spin concentration to about 0.5mL.
Concentrated solution is placed under nitrogen by 1.4 to be dried up, and adds acetonitrile+water (2+8, v/v) mixing of 2mL, constant volume after 0.2 μm of membrane filtration, obtains testing sample solution.
2, LC-Q-TOF/MS operating condition
Chromatographic condition: liquid chromatogram mobile phase A is ammonium acetate-0.1% formic acid-water of 5mM; Mobile phase B is acetonitrile; Gradient elution program is: 0min:1%B, 3min:30%B, 6min:40%B, 9min:40%B, 15min:60%B, 19min:90%B, 23min:90%B, 23.01min:1%B, runs 4min afterwards; Flow velocity is 0.4mL/min; Column temperature: 40 DEG C; Sample size: 10 μ L.
The mass spectrometric capillary voltage of Mass Spectrometry Conditions: Agilent6530LC-Q-TOF/MS: 4000V; Dry temperature: 325 DEG C; Dry gas stream amount 10L/min, sheath gas flow velocity 11L/min, sheath gas temperature is 325 DEG C; Atomization gas pressure 40psi, taper hole voltage 60V, fragmentation voltage 140V. Full scan karyoplasmic ratio ranges for 50-1600m/z, and adopts interior mark reference solution that instrument quality precision is carried out real time correction. By AgilentMassHunterWorkstationSoftware (versionB.05.00), mass spectrum testing result is gathered and process.
3 stem class vegetable (for Germinatus Phragmitis) Pesticide Residues detectings
3.1 measure sample solution under full scan pattern, are compared in detecting result and TOF data storehouse, draw 1 grade of TOF score, see Fig. 1.
3.2 meet the compound of requirement for score, set up two grades of acquisition methods in software, refer to Fig. 2.
3.3, under MS/MS pattern, rerun sample solution, it is thus achieved that sample fragment ion full scan data, it are compared with fragment ion information in second order ms picture library, obtain two grades of QTOF scores, see Fig. 3.
In 4 certain provincial capital's Germinatus Phragmitis sample, LC-Q-TOF/MS detects result
Gathering 4, the commercially available Germinatus Phragmitis sample of certain provincial capital, application LC-Q-TOF/MS technology carries out 544 kinds of pesticide residues detectings, LC-Q-TOF/MS technology 2 kinds of pesticide residues of detection, amounts to 2 frequencys, relates to 2, sample, and concrete outcome is in Table 2.
1544 kinds of pesticide LC-Q-TOF/MS detection parameters tables of table
In the Germinatus Phragmitis sample of table 2 somewhere, LC-Q-TOF/MS detects result
Claims (10)
1. one kind is used for detecting the method for detecting of 544 kinds of pesticide residues in stem class vegetable, it is characterized in that: by setting up TOF/MS data base and the Q-TOF/MS data base of 544 kinds of pesticide, the chromatographic mass spectrometry information of target compound is mated with the information retrieval of TOF/MS data base and Q-TOF/MS data base, achieve and compare without reference material, more than in agricultural product 500 kinds pesticide are carried out qualitative detecting and confirmation simultaneously; 544 kinds of described pesticide are the pesticide being numbered 1-544 in table 1.
2. a TOF/MS data base as claimed in claim 1, it is characterized in that: build data base by the following method: under TOF/MS pattern, measure every kind of pesticide standard thing retention time under specifying chromatograph Mass Spectrometry Conditions respectively, determine the ionized form under this compound ESI source, selection+H ,+NH4+ Na, and chemical formula, obtain the accurate mass number of every kind of compound parent ion, isotopic peak distribution and abundance ratio, the title of 544 kinds of pesticide, retention time, molecular formula, accurate mass number, isotopic peak distribution and abundance ratio information are imported in data base's csv file, forms TOF/MS data base.
3. a Q-TOF/MS data base as claimed in claim 1, it is characterized in that: build data base by the following method: the parent ion of 544 kinds of pesticide in data acquisition interface sequentially inputs TOF/MS data base as stated in claim 2, under Q-TOF/MS pattern, to 544 kinds of pesticide respectively under 8 collision energies, carry out fragment ion full scan mass spectrum collection, under informative 3-5 the different collision energies of ion, fragment ion full scan mass spectrum imports in database software file preferably wherein, and be associated with corresponding pesticide information, build up Q-TOF/MS data base.
4. method for detecting according to claim 1, it is characterised in that: the detecting for pesticide residues need not compare by pesticide standard thing.
5. method for detecting according to claim 1, it is characterised in that described method includes following procedure:
A () sample preparation: stem class vegetable sample extracts through acetate acetonitrile homogenate, after dehydration and centrifugal, concentration, then through Carbon/NH2Column purification, acetonitrile+toluene eluting residual pesticide, concentrated, filter after make and treat sample measuring liquid a;
B () chromatographic mass spectrometry detects: adopt the liquid chromatograph being furnished with reversed phase chromatographic column, under special color spectral condition, separates the pesticide treating in sample measuring liquid a; Adopt the LC-Q-TOF/MS-MS being furnished with electric spray ion source, under specific Mass Spectrometry Conditions, detect pesticide;
(c) TOF/MS data base's examination: call the TOF/MS data base as claimed in claim 2 set up in qualitative software, according to TOF/MS data base to target compound retrieval by header in mass spectrum testing result, search argument retention time deviation is defined to ± 0.5min, accurate mass deviation is defined to ± 10ppm, ionized form selection+H ,+NH4,+Na pattern, according to measured value and the deviation of theoretical value in TOF/MS data base of every kind of each key element of compound, provide retrieval matching score value, the compound of retrieval matching score value > 70, for doubtful detection compound;
D () Q-TOF/MS data base confirms: under TargetedMS/MS drainage pattern, input the parent ion of doubtful detection compound, retention time and best collision energy, treat sample measuring liquid a again to detect, spectrogram and spectrogram in Q-TOF/MS data base as claimed in claim 3 will be detected, the coupling confirmation when mirror image contrast, its matching score value > 70, namely confirm to detect this target compound.
6. method for detecting according to claim 5, it is characterised in that: in step (a), described stem class vegetable includes Germinatus Phragmitis.
7. method for detecting according to claim 5, it is characterised in that: in step (a), described Carbon/NH2Column purification step is as follows: at Carbon/NH2Post adds anhydrous sodium sulfate, first with acetonitrile+toluene mixture liquid drip washing SPE post, and discards effluent, when liquid level arrives the top of sodium sulfate, rapidly sample concentration liquid is transferred on decontaminating column, under connect new Cor Gigeriae Galli bottle graft and receive; Acetonitrile+toluene mixture liquid washing sample liquid bottle three times again, and cleaning mixture is moved in SPE post; Again with acetonitrile+toluene mixture liquid eluting pesticide, merge eluent in Cor Gigeriae Galli bottle.
8. method for detecting according to claim 5, it is characterized in that: in step (a), described concentrated, filter after make and treat that sample measuring liquid a refers to the eluent that claim 7 is obtained, spin concentration in 40 DEG C of water-baths, and concentrated solution is placed under nitrogen and dries up, add acetonitrile+formic acid water mixing, constant volume after membrane filtration, obtain testing sample solution a.
9. method for detecting according to claim 5, it is characterised in that: in step (b), described special color spectral condition refers to: liquid chromatogram mobile phase A is ammonium acetate-0.1% formic acid-water of 5mM; Mobile phase B is acetonitrile; Gradient elution program is: 0min:1%B, 3min:30%B, 6min:40%B, 9min:40%B, 15min:60%B, 19min:90%B, 23min:90%B, 23.01min:1%B, runs 4min afterwards; Flow velocity is 0.4mL/min; Column temperature: 40 DEG C; Sample size: 10 μ L.
10. method for detecting according to claim 5, it is characterised in that: in step (b), described specific Mass Spectrometry Conditions refers to: the mass spectrometric capillary voltage of Agilent6530LC-Q-TOF/MS: 4000V; Dry temperature: 325 DEG C; Dry gas stream amount 10L/min, sheath gas flow velocity 11L/min, sheath gas temperature is 325 DEG C; Atomization gas pressure 40psi, taper hole voltage 60V, fragmentation voltage 140V, full scan karyoplasmic ratio ranges for 50-1600m/z, and instrument quality precision is carried out real time correction by mark reference solution in adopting, and processed the collection of mass spectrum testing result by AgilentMassHunterWorkstationSoftware.
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| CN109557197A (en) * | 2017-04-17 | 2019-04-02 | 中国检验检疫科学研究院 | Agricultural chemical compound electronic identity database and detection method in edible agricultural product based on GC-Q-Orbitrap |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102221289A (en) * | 2011-05-26 | 2011-10-19 | 醴陵友立特种陶瓷有限公司 | Rotary kiln |
| CN105116064A (en) * | 2015-07-17 | 2015-12-02 | 中国农业科学院茶叶研究所 | Automatic sample pretreatment and detection method for detecting residual of multiple pesticides in tea leaves |
-
2015
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102221289A (en) * | 2011-05-26 | 2011-10-19 | 醴陵友立特种陶瓷有限公司 | Rotary kiln |
| CN105116064A (en) * | 2015-07-17 | 2015-12-02 | 中国农业科学院茶叶研究所 | Automatic sample pretreatment and detection method for detecting residual of multiple pesticides in tea leaves |
Non-Patent Citations (2)
| Title |
|---|
| 王慧君 等: "SPE和QuEChERS前处理方法结合LC-Q-TOF/MS检测苹果和番茄中282种农药残留对比研究", 《分析试验室》 * |
| 赵志远: "LC-Q-TOF/MS在果蔬中农药多残留快速筛查与确证分析中的应用", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109557197A (en) * | 2017-04-17 | 2019-04-02 | 中国检验检疫科学研究院 | Agricultural chemical compound electronic identity database and detection method in edible agricultural product based on GC-Q-Orbitrap |
| CN109557197B (en) * | 2017-04-17 | 2019-10-18 | 中国检验检疫科学研究院 | The method for building up and detection method of agricultural chemical compound electronic identity database in edible agricultural product based on GC-Q-Orbitrap |
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