CN107976490A - The remaining method of pesticide in one kind detection veterinary antibiotics - Google Patents
The remaining method of pesticide in one kind detection veterinary antibiotics Download PDFInfo
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- CN107976490A CN107976490A CN201710912245.5A CN201710912245A CN107976490A CN 107976490 A CN107976490 A CN 107976490A CN 201710912245 A CN201710912245 A CN 201710912245A CN 107976490 A CN107976490 A CN 107976490A
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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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Abstract
The invention discloses one kind to detect the remaining method of pesticide in veterinary antibiotics, this method carries out sample pre-treatments using improved QuEChERS kit methods, with the consumption of simple and quick, high-effective and labour-saving, favorable reproducibility and low reagent, low labor intensity, low cost, advantages of environment protection, examination, confirmatory analysis while coordinating UPLC MS/MS Qtrap detection methods to realize 59 kinds of pesticide residues at the same time, and then realize the remaining high throughput analysis detection of Multiple Pesticides.
Description
Technical field
The present invention discloses pesticide in the technical field for being related to Detecting Pesticide, more particularly to a kind of detection veterinary antibiotics
Remaining method.
Background technology
China is a large agricultural country, pesticide residue play the role of in agricultural production it is very important, from starting to use
Up to the present about thousands of kinds of pesticide residues are widely used in control of agricultural pest to the last century mid-term of pesticide residue.
Although current various countries are devoted to the pesticide residue of research efficiently, less toxic, the situation that pesticide residue excessively uses still ten
Divide universal.
After pesticide residue sprinkling, a part passes through plant leaf blade and root absorption in crop surface, soil；One
Divide with superfine particulate floating in an atmosphere, cause air pollution, dropped in soil and waters and made with sleet by water circulation
Into soil and water pollution.In addition, the natural calamity such as floods, earthquake, the unordered blowdown of pesticide residue factory, transport leakage, factory
The pesticide residue leakage that the accidents such as accident produce, causes water pollution, and then causes crop pollution, jeopardizes the people and is good for
Health.
Food is the root of life, is the irreplaceable valuable source of people's life.It is to cause food in view of pesticide residue
Poisoning or the main matter of contamination accident, it is imperative to research and develop the detection of pesticide residue, but existing detection method can only be realized
A kind of residue detection of pesticide residue, causes tested sample to need to reprocess, detects respectively, time and effort consuming, detection efficiency
It is low.
Therefore, detected while how researching and developing a kind of achievable Multiple Pesticides residual, particularly Multiple Pesticides are residual in sample
The examination stayed, confirmation are many pesticide residue analysis worker urgent problems to be solved.
The content of the invention
In consideration of it, the present disclosure provides one kind detect veterinary antibiotics in the remaining method of pesticide, with least solve with
Past detection method can only detect a kind of pesticide residue every time, cause tested sample to need to reprocess, detect respectively, take
Arduously, the problems such as detection efficiency is low.
Technical solution provided by the invention, specifically, a kind of remaining method of pesticide in detection veterinary antibiotics, its feature
It is, includes the following steps：
1) sample pre-treatments：
Weigh sample to be positioned in extracting tube, and add the acetate acetonitrile solution vortex oscillation that volumetric concentration is 1%, add
Homogeneous extracts after salt bag, centrifugation；
Take supernatant to be placed in purification pipe purified, concentrated, after constant volume, it is spare；
2) qualitative analysis：
Examination is carried out to the veterinary antibiotics sample to be detected Jing Guo sample pre-treatments with UPLC-MS/MS-Qtrap methods,
Using the enhancer ion scan function of liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer, MRM-IDA-EPI sieves are established
Checking method, obtains the MS of each target pesticide residue parent ion in veterinary antibiotics sample to be detected2Spectrogram, with given data storehouse
The standard MS of each pesticide residue2Spectrogram and the MS for obtaining each target pesticide residue parent ion in veterinary antibiotics sample to be detected2Spectrum
After figure is contrasted respectively, obtain similarity value, and match both fragments characteristic ion, so judge vegetables to be detected,
Whether contain target pesticide residue in fruit sample；
3) quantitative analysis：
Each trace standard of pesticide product are weighed respectively, prepare the mark of 10 μ g/mL respectively according to the similar principle selection solvent that mixes
Quasi- storing solution, it is spare；
It is dilute with blank sample matrix solution after being handled according to the sample-pretreating method in step 1) blank sample
Standard reserving solution is released, hybrid standard stock solution is made；
After hybrid standard stock solution constant volume, obtain blank sample matrix solution, compound concentration for 5ng/mL~
The series matching standard working solution of 500ng/mL；
Using liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer to series matching standard working solution and by sample
The veterinary antibiotics sample to be detected of product pre-treatment is measured respectively, is obtained using concentration as abscissa, the peak area of quota ion
For the drafting standard curve of ordinate, target pesticide residue in veterinary antibiotics sample to be detected is calculated with this standard curve
Content.
It is preferred that the pesticide is isofenphos, cadusafos, malathion, pyridaphethione, Profenofos, DDVP, kills snout moth's larva sulphur
Phosphorus, 2-dichloroethylk dimethyl phosphate, azinphos-methyl, parathion, chlorpyrifos-methyl, parathion-methyl, Azodrin, phosfolan, isazofos, Menite, kill
Flutter phosphorus, edifenphos, thimet, etrimfos, pirimiphos-methyl, acephatemet, Carbicron, demeton, disulfoton, Entex, quinoline
Sulphur phosphorus, fensulfothion, zolone, sulfotep, flolimat, fenamiphos, Rogor, diazinon, phosmet, phosphamidon, methyl sulphur ring
Phosphorus, Hostathion, imidacloprid, carbendazim, arprocarb, indoxacarb, tetrachlorvinphos, Benfuracard micro, carbofuran, Aldicarb sulfone, sulphur are double
Prestige, Methomyl, Aphox, Aldicarb, ficams, Propamocarb, carbaryl, Mobucin, diethofencarb, Bassa, methiocarb, fluorine amine
One or more in Cyano chrysanthemate.
Further preferably, the step 1) sample pre-treatments are specially：
Weigh sample 15.00g to be positioned in the 100mL extracting tubes for being loaded with ammonium sulfate, and add 15mL volumetric concentrations and be
1% acetate acetonitrile solution, in vortex oscillation 1min on eddy mixer, the salt in salt bag is all added in extracting tube,
Matter extracts 1min, and 3min is centrifuged with 3000r/min；
Supernatant is extracted in purifying in pipe with syringe clarifier, is inserted into plug, vortex oscillation 1min, filter membrane, correct amount
5mL scavenging solutions are taken, are concentrated to dryness under the conditions of 40 DEG C, add the methanol aqueous solution dissolved residue that 1.0mL volumes solubility is 20%,
0.2 μm of filter membrane is crossed, it is spare.
Further preferably, in the step 2) liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer Mass Spectrometry Conditions
For：
Ion gun：Electric spray ion source；
Scan pattern：Cation scans；
Drainage pattern：Multiple-reaction monitoring；
Information association scans：Intensity threshold 100cps；
Enhancer ion scan：50~1000Da of mass number acquisition range m/z；
Electron spray voltage：5500V；
Atomization gas pressure：50psi；
Gas curtain atmospheric pressure：30psi；
Assist gas pressure power：60psi；
Ion source temperature：550℃；
Remove cluster voltage：80V
Entrance potential：10V；
Collision gas energy：20、35、50V；
Extend collision energy：15V.
Further preferably, in the step 3) liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer chromatographic condition
For：
Chromatographic column：Waters BEH C18Reverse-phase chromatographic column；
Column temperature：40℃；
Sampling volume：10μL；
Flow velocity：0.3mL/min；
Mobile phase：Mobile phase A：Volumetric concentration be 0.2% aqueous formic acid, B：Volumetric concentration is 0.2% formic acid second
Nitrile solution, gradient elution program：0min, 95%A；1min, 95%A；10min, 60%A；15min, 5%A；18min, 5%A；
18.1min 95%A；23min, 95%A；
The Mass Spectrometry Conditions of liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer are in the step 3)：
Ion gun：Electric spray ion source；
Scan pattern：Cation scans；
Drainage pattern：Multiple-reaction monitoring；
Electron spray voltage：5500V；
Atomization gas pressure：50psi；
Gas curtain atmospheric pressure：30psi；
Assist gas pressure power：60psi；
Ion source temperature：550℃；
Remove cluster voltage：80V；
Entrance potential：10V；
Collision gas energy：20、35、50V；
Extend collision energy：15V.
Further preferably, the veterinary antibiotics sample to be detected be homogenized through tissue refiner celery, Chinese cabbage, west
Red persimmon, citrus, grape or apple.
The remaining method of pesticide in detection veterinary antibiotics provided by the invention, using improved QuEChERS kit sides
Method carry out sample pre-treatments, have simple and quick, high-effective and labour-saving, favorable reproducibility and the consumption of low reagent, low labor intensity, it is low into
Originally, advantages of environment protection, while coordinate UPLC-MS/MS-Qtrap detection methods to be sieved while can realizing 59 kinds of pesticide residues
Look into, confirmatory analysis, and then realize the remaining high throughput analysis detection of Multiple Pesticides.
The remaining method of pesticide in detection veterinary antibiotics provided by the invention, solve 59 kinds of pesticide residues extract at the same time,
The problem of purification, simplifies sample handling procedure, have efficiently, quick, cost is low, easy to operate, advantages of environment protection, only
It is good to change effect, high sensitivity, accuracy and precision meet the requirement of carbamate insecticides.
Brief description of the drawings
Attached drawing herein is merged in specification and forms the part of this specification, shows the implementation for meeting the present invention
Example, and for explaining the principle of the present invention together with specification.
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is attached drawing needed in technology description to be briefly described, it should be apparent that, for those of ordinary skill in the art
Speech, without creative efforts, can also obtain other attached drawings according to these attached drawings.
Fig. 1 is the MS that the present invention discloses suspicious peak in a kind of veterinary antibiotics to be detected that embodiment provides2Spectrogram；
Fig. 2 is the standard MS for pesticide residue-chlopyrifos that the present invention discloses embodiment offer2Spectrogram.
Embodiment
Here exemplary embodiment will be illustrated in detail, its example is illustrated in the accompanying drawings.Following description is related to
During attached drawing, unless otherwise indicated, the same numbers in different attached drawings represent the same or similar key element.Following exemplary embodiment
Described in embodiment do not represent and the consistent all embodiments of the present invention.On the contrary, they be only with it is such as appended
The example of the consistent method of some aspects being described in detail in claims, of the invention.
The present embodiment is achieved through the following technical solutions in veterinary antibiotics while detects organic phosphates, amino first
The method of totally 59 kinds of pesticide residues such as esters of gallic acid, specifically includes following steps：
Sample pre-treatments：
1st, sample extraction：
Accurately weigh sample 15.00g (being accurate to 0.01g) to be positioned in the 50mL extracting tubes for being loaded with ammonium sulfate, add
15mL volumetric concentrations are 1% acetate acetonitrile solution, after vortex oscillation 1min on eddy mixer, by QuEChERS kits
In salt bag open all add extracting tubes in, homogeneous extraction 1min, with 3000r/min centrifuge 3min；
2nd, sample purification：
Supernatant is extracted in purifying in pipe with syringe clarifier, is inserted into plug, vortex oscillation 1min, filter membrane, correct amount
5mL scavenging solutions are taken, 40 DEG C are concentrated to dryness, and add 1.0mL methanol aqueous solution (vol/vol methanols：Water=1：4) dissolved residue, mistake
0.2 μm of filter membrane, it is spare.
Qualitative analysis：
The legal measure of UPLC-MS/MS-Qtrap：Method is confirmed, it can be achieved that qualitative, quantitative using MRM-IDA-EPI examinations
It is disposable to complete, it can be applied to the analysis of complex matrices sample.
First using to be detected vegetables of the liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer Jing Guo sample pre-treatments,
Fruit sample carries out detection scanning, and whether the signal strength of system automatic decision detection scanning collection is more than in IDA condition settings
Every setting value, that is, judge whether chromatographic peak occur.When meeting preset level, system it is quick (<1ms) automatically switch to line
Property ion trap pattern, carry out enhancer ion scan (EPI), and obtain the high quality MS of parent ion obtained by detection scanning2Spectrogram,
With the standard MS of each pesticide residue in given data storehouse2Spectrogram is residual with obtaining each target pesticide in veterinary antibiotics sample to be detected
Stay the MS of parent ion2After spectrogram is contrasted respectively, similarity value is obtained, and matches both fragments characteristic ions, and then
Judge whether contain target pesticide residue in veterinary antibiotics sample to be detected.
Quantitative analysis：
Matrix matching standard working solution and detected sample are measured, using concentration as abscissa, quota ion
Peak area draws standard curve for ordinate, and the content of target pesticide residue in test sample is calculated with this standard curve.
Wherein, the chromatographic condition that liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer uses in quantitative analysis for：
Acquity Waters BEH C18Column (2.1 × 100mm, 1.7 μm), mobile phase A：0.2% formic acid that volumetric concentration is is water-soluble
Liquid；B：Volumetric concentration is 0.2% formic acid acetonitrile solution, and using gradient elution, condition of gradient elution is shown in Table 1, flow velocity 0.3mL/
Min, 40 DEG C of column temperature, 10 μ L of sampling volume.
1. liquid chromatogram gradient elution program of table
| Time (min) | Flow velocity (mL/min) | Mobile phase A % | Mobile phase B % |
| 0 | 0.3 | 95 | 5 |
| 1 | 0.3 | 95 | 5 |
| 10 | 0.3 | 60 | 40 |
| 15 | 0.3 | 5 | 95 |
| 18 | 0.3 | 5 | 95 |
| 18.1 | 0.3 | 95 | 5 |
| 23 | 0.3 | 95 | 5 |
Wherein, the Mass Spectrometry Conditions that liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer uses for：Ionization pattern：Electricity
Spray cation ESI (+)；Detection mode：Multiple-reaction monitoring scans (MRM)；Electron spray voltage (IS)：5500V；Atomization gas pressure
(GS1)：50psi；Gas curtain atmospheric pressure (CUR)：30psi；Assist gas pressure power (GS2)：60psi；Ion source temperature (TEM)：550
℃；Remove cluster voltage (DP)：80V；Entrance potential (EP)：10V.
The retention time and mass spectrum acquisition parameter of 59 kinds of pesticide residues are shown in Table 2.
The mass spectrometry parameters of 2. pesticide residue to be measured of table
* quota ion.
The remaining method of pesticide compared with prior art, has following in the detection veterinary antibiotics that the present embodiment provides
Excellent results：
(1) the present embodiment establishes a kind of UPLC-MS/MS-Qtrap for analyzing 59 kinds of pesticide residues in veterinary antibiotics
Examination assay method, is detected while realizing variety classes pesticide residue.
(2) the present embodiment is realized 59 kinds of pesticide residues and is carried at the same time using improved QuEChERS methods processing sample
Take, purify, there is easy, efficient, quick, advantages of environment protection.
(3) the present embodiment uses improved QuEChERS kit methods, wherein, improved QuEChERS kit sides
Method, is inherently added with ammonium sulfate in extracting tube, and is additionally provided with for individually packing of being added in extraction step
Salt bag and needle cylinder type filter membrane filter is used in purifying step.Compared with traditional QuEChERS methods, sulphur in extracting tube
Sour ammonium can ensure that the pH value of solution as the Acetic acid-sodium acetate buffer system result in former method, only need to add acetonitrile one
The common solution of kind.Proprietary salt bag, spills when avoiding directly weighing, importing, easy to operate.Avoid in extraction process first
Anhydrous magnesium sulfate and inorganic salts are added, magnesium sulfate lumps and caloric value is excessive, easily causes the volatilization loss of volatile determinand.
Needle cylinder type filter membrane filter cleaning mode, without centrifugation, you can loading is analyzed, and easy to operate, cost is low, good purification.
(4) the present embodiment is good using selectivity, and the UPLC-MSMS-Qtrap methods detection of strong antijamming capability, stablizes
Property good, high sensitivity, accuracy and precision meets the requirement of carbamate insecticides, while has the function of examination, confirmation,
It can avoid the detection of false positive sample.MRM-IDA-EPI functions are selected, relevant criterion is set in IDA, connect liquid
Phase, inputs the acquisition condition of liquid phase, carries out data acquisition, once the signal strength of MRM collections exceedes criterion value, will touch
Send out EPI and carry out enhancer ion scan collection MS2Figure, its suspicious peak is opened (by retention time, peak shape etc. to the collection of illustrative plates of collection
Judge) MS2Figure, carries out the operations such as known spectrum library searching, is contrasted with the mass spectrometric data for composing storehouse, obtain corresponding coupling number
According to, to carry out judging remaining detection, and then realize rapid screening and confirmation purpose.
(5) detection limit of the present embodiment and quantitative limit are low, method high sensitivity, and the rate of recovery is stablized, and reproduction is acted charitably, met
The requirement of multi-residue analysis.
Specific embodiment
The present invention is further illustrated with specific embodiment below, but is not intended to limit the invention
Protection domain.
59 kinds are carried out respectively to six kinds of celery, Chinese cabbage, tomato, citrus, grape, apple samples using the above method below
The measure of persticide residue and suspicious peak examination.
1. instrument and reagent
Key instrument：ACQUITYTMUltra Performance Liquid Chromatography instrument (Waters, US)；AB Sciex QTrap 5500
Quadrupole rod-ion trap mass spectrometer (American AB company)；QuEChERS kits (Shandong Province of China green grass or young crops into Instrument Ltd.).
Main agents：The purity of 59 kinds of standard items >=98.0%, purchased from Inst. of Environment Protection ＆ Scientific Research Monitor, Ministry of Agric (my god
Tianjin), standard items production firm and the research institution such as State center for standard matter.
2. standard solution is prepared：
Prepare standard reserving solution：Weigh that each standard items are appropriate respectively, suitable solvent (first is selected according to the similar principle that mixes
Alcohol, acetonitrile etc.) standard reserving solution of about 10 μ g/mL is formulated as, it is placed in brown bottle and is saved backup at -20 DEG C or 4 DEG C.
Prepare matrix matching standard working solution：In order to eliminate influence of the matrix effect to quantitative determination, blank sample is used
Solution dilutes hybrid standard stock solution.It is molten with blank sample matrix to blank sample by following identical pretreatment mode processing
Liquid dilutes hybrid standard stock solution, and constant volume obtains blank sample matrix solution, compound concentration 5ng/mL, 20ng/mL,
The series matching standard working solution of 50ng/mL, 100ng/mL, 250ng/mL, 500ng/mL.
3. chromatograph mass spectrum analysis condition：
Chromatographiccondition：
Waters BEH C18Reverse-phase chromatographic column；Mobile phase A：Volumetric concentration be 0.2% aqueous formic acid, B：Volume is dense
Spend the formic acid acetonitrile solution for 0.2%；Using gradient elution, flow velocity 0.3mL/min, gradient elution program：0min, 95%A；
1min, 95%A；10min, 60%A；15min, 5%A；18min, 5%A；18.1min 95%A；23min, 95%A；Column temperature 40
℃；10 μ L of sampling volume.
Mass spectral analysis condition：
Ion gun：Electric spray ion source；Scan pattern：Cation scans；Drainage pattern：Multiple-reaction monitoring (MRM)；EFI
Mist voltage (IS)：5500V；Atomization gas pressure (GS1)：50psi；Gas curtain atmospheric pressure (CUR)：30psi；Assist gas pressure power (GS2)：
60psi；Ion source temperature (TEM)：550℃；Remove cluster voltage (DP)：80V；Entrance potential (EP)：10V.Wherein, 59 kinds of pesticides are residual
The retention time and mass spectrum acquisition parameter stayed are shown in Table 2.
4. sample-pretreating method：
Sample extraction：The accurate sample 15.00g (being accurate to 0.01g) for weighing homogeneous extracts in the 50mL for being loaded with ammonium sulfate
Take in pipe, add the acetate acetonitrile solution that 15mL volumetric concentrations are 1%, will after vortex oscillation 1min on eddy mixer
Salt bag, which is opened, in QuEChERS kits all adds in extracting tube, and homogeneous extraction 1min, 3min is centrifuged with 3000r/min.
Sample purification：Supernatant is extracted in purifying in pipe with syringe clarifier, is inserted into plug, vortex oscillation 1min, filtering
Film, accurate to measure 5mL scavenging solutions, 40 DEG C are concentrated to dryness, and add 1.0mL methanol aqueous solution (vol/vol methanols：Water=1：4) dissolve
Residue, crosses 0.2 μm of filter membrane, spare.
5. quantitative analysis and the efficiency evaluation of method：
Linearly, detection limit and quantitative limit：The matrix matching standard solution of series concentration is measured, using concentration as horizontal seat
Mark, the peak area of quota ion draw standard curve for ordinate.Then test sample is measured, it is residual obtains various pesticides
The peak area of the quota ion stayed, substitute into standard curve, calculate 59 kinds of pesticide residues content.Added not into blank sample
The standard solution of homogenous quantities, the detection limit and quantitative limit of method are calculated with 3 times of signal-to-noise ratio and 10 times of signal-to-noise ratio respectively.59 kinds of agricultures
The remaining detection limit of medicine is between 0.01~0.81 μ g/kg, and quantitative limit is between 0.03~2.67 μ g/kg, the line of each compound
Property regression equation, detection limit, quantitative limit are shown in Table 3.
The equation of linear regression of 3. pesticide residue of table, correlation coefficient r, detection limit, quantitative limit
Accuracy and precision：Recovery testu, each concentration level are done in 5,10,50 μ g/kg, tri- concentration levels
6 parallel samples are done, carry out sample pre-treatments and Instrumental Analysis respectively, and according to adding scalar sum measured value to calculate the rate of recovery.Vegetable
The average recovery rate of 59 kinds of pesticide residues and relative standard deviation (RSD) are shown in Table 4 in dish fruit.As can be seen from Table 4,59 kinds of pesticides
The average recovery rate in six kinds of celery, Chinese cabbage, tomato, citrus, grape, apple samples is remained in 60.0%~111.4%
Between, relative standard deviation meets the requirement of multi-residue analysis between 1.2%~18.7%.
The rate of recovery and relative standard deviation (n=6) in 4. veterinary antibiotics sample of table under different pitch-based spheres
6. suspicious peak qualitatively screening analysis citing
High-throughout unknown material rapid screening is carried out using Analyst softwares.Actual sample is detected, in MRM scan patterns
Lower discovery suspected target thing, by comparing fragment in actual sample testing result and database under three kinds of energy, abundance ratio, protecting
The information such as time are stayed to carry out compound confirmation.
Mass Spectrometry Conditions are：Ion gun：Electric spray ion source；Scan pattern：Cation scans；Drainage pattern：More reaction prisons
Survey (MRM)；Information association scans (IDA)：Intensity threshold 100cps；Enhancer ion scan (EPI), mass number acquisition range m/
50~1000Da of z；Electron spray voltage (IS)：5500V；Atomization gas pressure (GS1)：50psi；Gas curtain atmospheric pressure (CUR)：
30psi；Assist gas pressure power (GS2)：60psi；Ion source temperature (TEM)：550℃；Remove cluster voltage (DP)：80V；Entrance potential
(EP)：10V；Collision gas energy (CE)：20、35、50V；Extend collision energy (CES)：15V.
Select MS2Spectrogram and maximize figure, right click mouse clicks on Search Library, and software can be given automatically
Go out the result of examination.Fit values (matching value) be obtained after being contrasted with standard substance spectrogram to suspicious specimen spectrogram it is similar
Angle value, full marks 100, the possibility of the higher explanation compound of the bigger confidence level of Fit values are bigger；Revfit values (anti-phase matching value)
It is the similarity value obtained after being contrasted with suspicious specimen spectrogram and standard substance spectrogram；Purity values (Reinheitszahl) are comprehensive
The numerical value that first two result obtains.Fig. 1 is the MS at suspicious peak in sample2Spectrogram, Fig. 2 are the MS of chlopyrifos standard items2Spectrogram, from
For table 5 it will be evident that storehouse Chlorpyrifos (Chlorpyrifos) score is high, arrangement is most forward, and fragments characteristic ion all with
Matched in sample, thus can determine whether to contain chlopyrifos in this sample, further increased the confidence to result confirmation, avoid vacation
The generation of positive findings.
Table 5.
| Name | Formula | Molecularweight(Da) | Fit | RevFit | Purity | CE | |
| 1 | Chlorpyrifos | C9H11Cl3NO3PS | 348.9000 | 84.418 | 70.480 | 66.438 | 35.000 |
| 2 | Chlorpyrifos | C9H11Cl3NO3PS | 348.9000 | 74.588 | 84.052 | 65.038 | 35.000 |
| 3 | Chlorpyrifos | C9H11Cl3NO3PS | 348.9000 | 85.133 | 62.197 | 55.456 | 50.000 |
| 4 | Chlorpyrifos | C9H11Cl3NO3PS | 348.9000 | 61.269 | 68.891 | 47.973 | 20.000 |
| 5 | Dicloxacillin | C19H17Cl2N3O5S | 469.0000 | 20.855 | 38.684 | 8.067 | 50.000 |
| 6 | Cadusafos | C10H23O2PS2 | 270.4000 | 56.461 | 13.265 | 7.912 | 50.000 |
| 7 | Sulprofos | C12H19O2PS3 | 322.3000 | 53.210 | 7.694 | 7.252 | 50.000 |
| 8 | Ethoprophos | C8H19O2PS2 | 242.0000 | 49.436 | 13.264 | 6.557 | 50.000 |
| … |
Those skilled in the art will readily occur to the present invention its after considering specification and putting into practice invention disclosed herein
Its embodiment.This application is intended to cover the present invention any variations, uses, or adaptations, these modifications, purposes or
Person's adaptive change follows the general principle of the present invention and including undocumented common knowledge in the art of the invention
Or conventional techniques.Description and embodiments are considered only as exemplary, and true scope and spirit of the invention are by following
Claim is pointed out.
It should be appreciated that the invention is not limited in the precision architecture for being described above and being shown in the drawings, and
And various modifications and changes may be made without departing from the scope thereof.The scope of the present invention is only limited by appended claim.
Claims (6)
1. the remaining method of pesticide in one kind detection veterinary antibiotics, it is characterised in that include the following steps：
1) sample pre-treatments：
Weigh sample to be positioned in extracting tube, and add the acetate acetonitrile solution vortex oscillation that volumetric concentration is 1%, add salt bag
Homogeneous extracts afterwards, centrifugation；
Take supernatant to be placed in purification pipe purified, concentrated, after constant volume, it is spare；
2) qualitative analysis：
Examination is carried out to the veterinary antibiotics sample to be detected Jing Guo sample pre-treatments with UPLC-MS/MS-Qtrap methods, is used
The enhancer ion scan function of liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer, establishes MRM-IDA-EPI examination sides
Method, obtains the MS of each target pesticide residue parent ion in veterinary antibiotics sample to be detected2Spectrogram, with each agriculture in given data storehouse
The remaining standard MS of medicine2Spectrogram and the MS for obtaining each target pesticide residue parent ion in veterinary antibiotics sample to be detected2Spectrogram point
After not contrasted, similarity value is obtained, and matches both fragments characteristic ions, and then judges veterinary antibiotics to be detected
Whether contain target pesticide residue in sample；
3) quantitative analysis：
Each trace standard of pesticide product are weighed respectively, prepare the standard storage of 10 μ g/mL respectively according to the similar principle selection solvent that mixes
Standby liquid, it is spare；
After being handled according to the sample-pretreating method in step 1) blank sample, diluted and marked with blank sample matrix solution
Quasi- storing solution, is made hybrid standard stock solution；
After hybrid standard stock solution constant volume, blank sample matrix solution is obtained, compound concentration is 5ng/mL~500ng/mL
Series matching standard working solution；
Using liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer to series matching standard working solution and by before sample
The veterinary antibiotics sample to be detected of processing is measured respectively, is obtained using concentration as abscissa, the peak area of quota ion is vertical
The drafting standard curve of coordinate, the content of target pesticide residue in veterinary antibiotics sample to be detected is calculated with this standard curve.
2. the remaining method of pesticide in veterinary antibiotics is detected according to claim 1, it is characterised in that the pesticide is different
Willow phosphorus, cadusafos, malathion, pyridaphethione, Profenofos, DDVP, fenifrothion, 2-dichloroethylk dimethyl phosphate, azinphos-methyl, parathion, first
The phonetic sulphur of base chlopyrifos, parathion-methyl, Azodrin, phosfolan, isazofos, Menite, methidathion, edifenphos, thimet, second
Phosphorus, pirimiphos-methyl, acephatemet, Carbicron, demeton, disulfoton, Entex, quinalphos, fensulfothion, zolone, sulfotep,
It is flolimat, fenamiphos, Rogor, diazinon, phosmet, phosphamidon, phosfolan_methyl, Hostathion, imidacloprid, carbendazim, residual
Kill prestige, indoxacarb, tetrachlorvinphos, Benfuracard micro, carbofuran, Aldicarb sulfone, thiodicarb, Methomyl, Aphox, Aldicarb, Evil
One or more in worm prestige, Propamocarb, carbaryl, Mobucin, diethofencarb, Bassa, methiocarb, taufluvalinate.
3. the remaining method of pesticide in veterinary antibiotics is detected according to claim 1, it is characterised in that the step 1) sample
Product pre-treatment is specially：
Weigh sample 15.00g to be positioned in the 100mL extracting tubes for being loaded with ammonium sulfate, and it is 1% to add 15mL volumetric concentrations
Acetate acetonitrile solution, in vortex oscillation 1min on eddy mixer, the salt in salt bag is all added in extracting tube, homogeneous extraction
1min, 3min is centrifuged with 3000r/min；
Supernatant is extracted in purifying in pipe with syringe clarifier, is inserted into plug, vortex oscillation 1min, filter membrane, accurately measures 5mL
Scavenging solution, is concentrated to dryness under the conditions of 40 DEG C, adds the methanol aqueous solution dissolved residue that 1.0mL volumes solubility is 20%, crosses 0.2
μm filter membrane, it is spare.
4. the remaining method of pesticide in veterinary antibiotics is detected according to claim 1, it is characterised in that in the step 2)
The Mass Spectrometry Conditions of liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer are：
Ion gun：Electric spray ion source；
Scan pattern：Cation scans；
Drainage pattern：Multiple-reaction monitoring；
Information association scans：Intensity threshold 100cps；
Enhancer ion scan：50~1000Da of mass number acquisition range m/z；
Electron spray voltage：5500V；
Atomization gas pressure：50psi；
Gas curtain atmospheric pressure：30psi；
Assist gas pressure power：60psi；
Ion source temperature：550℃；
Remove cluster voltage：80V
Entrance potential：10V；
Collision gas energy：20、35、50V；
Extend collision energy：15V.
5. the remaining method of pesticide in veterinary antibiotics is detected according to claim 1, it is characterised in that in the step 3)
The chromatographic condition of liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer is：
Chromatographic column：Waters BEH C18Reverse-phase chromatographic column；
Column temperature：40℃；
Sampling volume：10μL；
Flow velocity：0.3mL/min；
Mobile phase：Mobile phase A：Volumetric concentration be 0.2% aqueous formic acid, B：The formic acid acetonitrile that volumetric concentration is 0.2% is molten
Liquid, gradient elution program：0min, 95%A；1min, 95%A；10min, 60%A；15min, 5%A；18min, 5%A；
18.1min 95%A；23min, 95%A；
The Mass Spectrometry Conditions of liquid chromatography-tandem quadrupole rod linear ion trap mass spectrometer are in the step 3)：
Ion gun：Electric spray ion source；
Scan pattern：Cation scans；
Drainage pattern：Multiple-reaction monitoring；
Electron spray voltage：5500V；
Atomization gas pressure：50psi；
Gas curtain atmospheric pressure：30psi；
Assist gas pressure power：60psi；
Ion source temperature：550℃；
Remove cluster voltage：80V；
Entrance potential：10V；
Collision gas energy：20、35、50V；
Extend collision energy：15V.
6. the remaining method of pesticide in veterinary antibiotics is detected according to claim 1, it is characterised in that the vegetable to be detected
Dish, fruit sample are celery, Chinese cabbage, tomato, citrus, grape or the apple being homogenized through tissue refiner.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108318643A (en) * | 2018-05-12 | 2018-07-24 | 北京逸智联科技有限公司 | Pesticide residue determination method and detection device |
| CN108572227A (en) * | 2018-06-22 | 2018-09-25 | 苏州仁益生物科技有限公司 | A kind of vegetables detection remaining method of carbendazim of agricultural |
| CN109870529A (en) * | 2019-03-22 | 2019-06-11 | 生态环境部南京环境科学研究所 | Detect the pre-treating method and detection method of the high dead tick of pigment substrate intoxication |
| CN109870530A (en) * | 2019-03-22 | 2019-06-11 | 生态环境部南京环境科学研究所 | The detection method of the high dead tick of pigment substrate intoxication |
| CN110057928A (en) * | 2019-02-27 | 2019-07-26 | 中国热带农业科学院农产品加工研究所 | A kind of ultra performance liquid chromatography tandem mass spectrometry detecting captan residual quantity in tomato |
| CN110887926A (en) * | 2019-12-19 | 2020-03-17 | 浙江省农业科学院 | Sample pretreatment method for determining residual quantity of various pesticides in dendrobium officinale and detection method thereof |
| CN112557529A (en) * | 2020-11-24 | 2021-03-26 | 湖南省农业生物技术研究所 | Ultra-high performance liquid chromatography tandem mass spectrometry detection method for epoxiconazole and carbendazim |
| CN112730678A (en) * | 2020-12-24 | 2021-04-30 | 锐德检测技术(天津)有限公司 | Method for simultaneously detecting and analyzing various pesticide residues in fruits and vegetables |
| CN115372492A (en) * | 2021-12-16 | 2022-11-22 | 广东微谱检测技术有限公司 | Method for detecting residual quantity of thiophosphoryl methyl by LC-MS/MS and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103698195A (en) * | 2013-12-24 | 2014-04-02 | 烟台青云仪器设备有限公司 | QuEchERS method package for efficiently and homogeneously extracting residues of pesticide and veterinary drug |
| CN106546693A (en) * | 2016-10-25 | 2017-03-29 | 烟台大学 | A kind of method of portable imaging method detection sulfa drug residue and application thereof |
-
2017
- 2017-09-29 CN CN201710912245.5A patent/CN107976490A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103698195A (en) * | 2013-12-24 | 2014-04-02 | 烟台青云仪器设备有限公司 | QuEchERS method package for efficiently and homogeneously extracting residues of pesticide and veterinary drug |
| CN106546693A (en) * | 2016-10-25 | 2017-03-29 | 烟台大学 | A kind of method of portable imaging method detection sulfa drug residue and application thereof |
Non-Patent Citations (6)
| Title |
|---|
| 刘柱 等: "同位素稀释-超高效液相色谱-线性离子阱串联质谱分析谷物及其制品中呕吐毒素及其衍生物和代谢物", 《分析化学》 * |
| 张玉芬 等: "《农药残留检测与安全性评价》", 31 March 2013, 黑龙江大学出版社 * |
| 蔡勤仁 等: "液相色谱-串联质谱结合谱库检索法同时测定猪组织中12种类固醇激素", 《色谱》 * |
| 赵颖 等: "蔬菜和水果中59种农药残留同时检测-超高效液相色谱串联质谱法", 《广东化工》 * |
| 金雁 等: "EPI扫描模式筛查与鉴定畜禽产品中多种抗生素残留", 《重庆理工大学学报（自然科学）》 * |
| 黎娟 等: "高效液相色谱-四极杆/离子阱质谱确证测定面粉及面制品中氨基脲", 《分析测试学报》 * |
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|---|---|---|---|---|
| CN108318643A (en) * | 2018-05-12 | 2018-07-24 | 北京逸智联科技有限公司 | Pesticide residue determination method and detection device |
| CN108318643B (en) * | 2018-05-12 | 2020-12-25 | 曼哈格检测技术股份有限公司 | Method and device for detecting pesticide residue |
| CN108572227A (en) * | 2018-06-22 | 2018-09-25 | 苏州仁益生物科技有限公司 | A kind of vegetables detection remaining method of carbendazim of agricultural |
| CN110057928A (en) * | 2019-02-27 | 2019-07-26 | 中国热带农业科学院农产品加工研究所 | A kind of ultra performance liquid chromatography tandem mass spectrometry detecting captan residual quantity in tomato |
| CN109870529A (en) * | 2019-03-22 | 2019-06-11 | 生态环境部南京环境科学研究所 | Detect the pre-treating method and detection method of the high dead tick of pigment substrate intoxication |
| CN109870530A (en) * | 2019-03-22 | 2019-06-11 | 生态环境部南京环境科学研究所 | The detection method of the high dead tick of pigment substrate intoxication |
| CN110887926A (en) * | 2019-12-19 | 2020-03-17 | 浙江省农业科学院 | Sample pretreatment method for determining residual quantity of various pesticides in dendrobium officinale and detection method thereof |
| CN110887926B (en) * | 2019-12-19 | 2022-02-25 | 浙江省农业科学院 | Sample pretreatment method for determining residual quantity of various pesticides in dendrobium officinale and detection method thereof |
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