CN104678023B - A kind of GC-MS/MS measures the method for fluorine ether bacterium amide residual in fruit and vegerable - Google Patents
A kind of GC-MS/MS measures the method for fluorine ether bacterium amide residual in fruit and vegerable Download PDFInfo
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 title claims abstract description 126
- 229910052731 fluorine Inorganic materials 0.000 title claims abstract description 65
- 241000894006 Bacteria Species 0.000 title claims abstract description 64
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 title claims abstract description 64
- 239000011737 fluorine Substances 0.000 title claims abstract description 64
- 150000001408 amides Chemical class 0.000 title claims abstract description 62
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 22
- 235000013399 edible fruits Nutrition 0.000 title claims abstract description 16
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 48
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000001819 mass spectrum Methods 0.000 claims abstract description 7
- 239000000523 sample Substances 0.000 claims description 55
- 239000000243 solution Substances 0.000 claims description 26
- 239000007788 liquid Substances 0.000 claims description 24
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 20
- 239000012224 working solution Substances 0.000 claims description 19
- 150000002500 ions Chemical class 0.000 claims description 16
- 238000000605 extraction Methods 0.000 claims description 15
- 239000007789 gas Substances 0.000 claims description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 12
- 239000012046 mixed solvent Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 7
- 239000012496 blank sample Substances 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 235000013311 vegetables Nutrition 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 5
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims description 5
- 238000000611 regression analysis Methods 0.000 claims description 5
- 239000001632 sodium acetate Substances 0.000 claims description 5
- 235000017281 sodium acetate Nutrition 0.000 claims description 5
- 230000005540 biological transmission Effects 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 3
- 238000004364 calculation method Methods 0.000 claims description 3
- 239000012159 carrier gas Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000002552 multiple reaction monitoring Methods 0.000 claims description 3
- 239000011159 matrix material Substances 0.000 claims description 2
- 230000010355 oscillation Effects 0.000 claims description 2
- 230000002000 scavenging effect Effects 0.000 claims description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims 2
- 229910052786 argon Inorganic materials 0.000 claims 1
- 239000000575 pesticide Substances 0.000 abstract description 22
- 238000001514 detection method Methods 0.000 abstract description 13
- 238000011084 recovery Methods 0.000 abstract description 7
- 235000013305 food Nutrition 0.000 abstract description 6
- 239000000758 substrate Substances 0.000 abstract description 6
- 238000005516 engineering process Methods 0.000 abstract description 5
- 230000008901 benefit Effects 0.000 abstract description 3
- 238000011161 development Methods 0.000 abstract description 3
- 238000010812 external standard method Methods 0.000 abstract description 2
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical group CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 abstract description 2
- UIDUKLCLJMXFEO-UHFFFAOYSA-N propylsilane Chemical compound CCC[SiH3] UIDUKLCLJMXFEO-UHFFFAOYSA-N 0.000 abstract 2
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 abstract 1
- 239000006185 dispersion Substances 0.000 abstract 1
- 240000004160 Capsicum annuum Species 0.000 description 5
- 235000002567 Capsicum annuum Nutrition 0.000 description 5
- 230000018044 dehydration Effects 0.000 description 5
- 238000006297 dehydration reaction Methods 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 5
- 125000001153 fluoro group Chemical group F* 0.000 description 4
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 4
- 238000002203 pretreatment Methods 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 239000012086 standard solution Substances 0.000 description 4
- 238000004949 mass spectrometry Methods 0.000 description 3
- 244000052769 pathogen Species 0.000 description 3
- 230000001717 pathogenic effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000004885 tandem mass spectrometry Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 2
- 238000003705 background correction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- DNJKFZQFTZJKDK-UHFFFAOYSA-N fluopimomide Chemical compound FC1=C(F)C(OC)=C(F)C(F)=C1C(=O)NCC1=NC=C(C(F)(F)F)C=C1Cl DNJKFZQFTZJKDK-UHFFFAOYSA-N 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
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- 230000008569 process Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 1
- 241000193738 Bacillus anthracis Species 0.000 description 1
- KXDAEFPNCMNJSK-UHFFFAOYSA-N Benzamide Chemical class NC(=O)C1=CC=CC=C1 KXDAEFPNCMNJSK-UHFFFAOYSA-N 0.000 description 1
- 241000123650 Botrytis cinerea Species 0.000 description 1
- 241000609455 Corynespora cassiicola Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 0.000 description 1
- 244000307700 Fragaria vesca Species 0.000 description 1
- 235000016623 Fragaria vesca Nutrition 0.000 description 1
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 1
- 241000223218 Fusarium Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241001330975 Magnaporthe oryzae Species 0.000 description 1
- 241000220225 Malus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000222291 Passalora fulva Species 0.000 description 1
- 241000233616 Phytophthora capsici Species 0.000 description 1
- 241000233622 Phytophthora infestans Species 0.000 description 1
- 241000813090 Rhizoctonia solani Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
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- 238000001727 in vivo Methods 0.000 description 1
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- 244000144972 livestock Species 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
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- 238000011160 research Methods 0.000 description 1
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- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of GC MS/MS and measure the method for fluorine ether bacterium amide residual in fruit and vegerable, extracting the fluorine ether bacterium amide of residual in sample with acetonitrile or the acetonitrile solution homogenizing containing 1% acetic acid, extracting solution is through ethylenediamine N propyl silane (PSA) and octadecylsilane Bonded Phase (C18) substrate dispersion purify after, gas chromatogram tandem mass spectrum (GC MS/MS) detect, use without pesticide to be measured vehicle solution set up correction standard curve, quantified by external standard method.This method average recovery rate is 89.1%～95.0%, and average relative standard's deviation (RSD) is 4.0%～5.3%, and detection limit is less than 1.02 μ g/kg, has easy and simple to handle, quick, highly sensitive, reproducible, qualitative, quantitative advantage accurately." uniform limit " technology requirement of 0.01 mg/kg residue limits can be met, for ensureing that our people's food safety, export abroad trade sound development provide strong technical support.
Description
Technical field
The present invention relates to a kind of GC-MS/MS and measure the method for fluorine ether bacterium amide residual in fruit and vegerable, more specifically use gas
Phase chromatographic tandem mass spectrum (GC-MS/MS) qualitative, quantitative measures the method for the fluorine ether bacterium amide content of residual in vegetable and fruit,
Belong to the determination techniques field of persticide residue.
Background technology
Fluorine ether bacterium amide (LH-2010A) is that Shandong United Pesticide Industry Co., Ltd. is new in the one of innovation synthesis in 2010
Type fluorine-containing Benzoylamide series bactericidal agent, chemical name is: N-(3-chloro-5-(trifluoromethyl) pyridine-2-methyl)-2,3,5,6-tetrafluoros
-4-methoxy benzamide, structural formula is:
Containing 7 fluorine atoms in fluorine ether bacterium amide structure formula.The C-F key bond energy that fluorine atom is formed is much larger than c h bond, hence it is evident that
Add stability and the physiologically active of organofluorine compound.Fluorinated organic compound has higher fat-soluble and hydrophobicity, energy
Enough promote that it absorbs and transmission in vivo, strengthen the binding ability with organism, make the physiological action of organism change.
Fluoro-containing pesticide has such effect equally, and pathogen or the suppression of insect or toxic action are also greatly improved by fluoro-containing pesticide.According to grinding
Studying carefully discovery, fluorine ether bacterium amide is to cotton rhizoctonia solani, botrytis cinerea, apple anthrax bacteria, Fulvia fulva, Fructus Mali pumilae
Target spot pathogen, phytophthora infestans, Leaf Spot Caused by Corynespora cassiicola on Cucumber bacterium, phytophthora blight of pepper, Strawberry Fusarium Wilt and Pyricularia oryzae 10 kinds are planted
The virulence of thing pathogen is the highest, it is seen then that fluorine ether bacterium amide sterilizes more wide spectrum, can be as a kind of New-type wide-spectrum type antibacterial
Using, development prospect is wide.
Along with fluorine ether bacterium amide registration, promote and use, relevant fluorine ether bacterium amide Residue Degradation is dynamically and final residue etc.
The research of environmental behaviour certainly will increase, meanwhile, if European Union, Japan and other countries regulation field as China's main exit market make
Do not register in this country with pesticide, when not formulating corresponding residue limits standard, be exported to the food agricultural product bag of its country
Include residue limits in the animal derived foods such as livestock meat and all carry out " uniform limit " of 0.01mg/L.
Up to now, have no and be related to the report of fluorine ether bacterium amide residues detection method in food agricultural product both at home and abroad, use
Gas chromatogram tandem mass spectrum (GC-MS/MS) analyze food Residual Pesticides in Farm Produce have quick, easy, highly sensitive,
The advantages such as selectivity is strong, can realize the multi-residue analysis of hundreds of kind of pesticide, hence set up simplicity, quick, accurate, durable, energy
In accurate qualitative and quantitative analysis vegetable and fruit, the GC-MS/MS detection method of fluorine ether bacterium amide residual quantity is significant.
Summary of the invention
It is an object of the invention to provide a kind of GC-MS/MS and measure the method for fluorine ether bacterium amide residual in fruit and vegerable.
For realizing object above, the technical solution adopted in the present invention is: a kind of GC-MS/MS measures fluorine ether bacterium acyl in fruit and vegerable
The method of amine residual, comprises the steps:
(1) extract
Weigh sample in tool plug centrifuge tube, add acetonitrile or the acetonitrile solution homogenizing containing 1% acetic acid extracts 1min, add sodium chloride or second
One in acid sodium and anhydrous magnesium sulfate, centrifugal after vibration.
(2) purify
Pipette sample extracting solution supernatant in centrifuge tube, add Dispersive solid phase extraction agent, vortex oscillation, centrifugal, draw certain
After amount scavenging solution nitrogen dries up, dissolve constant volume with the acetone that volume ratio is 1/1/normal hexane mixed solvent, after crossing film, treat gas chromatogram
Tandem mass spectrum (GC-MS/MS) detects.
(3) preparation of standard working solution
When the same kind matrix blank sample of not fluorine-containing ether bacterium amide is processed by above-mentioned steps (1), (2), obtain sample extraction and purify
Residue, obtains vehicle solution after adding the mixing of appropriate solvent vortex, with the fluorine ether bacterium amide of this solution at least 3 concentration of preparation
Series hybrid standard working solution.
(4) gas chromatogram tandem mass spectrometry (GC-MS/MS) measures
The standard working solution of each Concentraton gradient in step (3) is carried out GC-MS/MS mensuration, with the chromatographic peak area of standard working solution
Its respective concentration is carried out regression analysis, obtains standard working curve；Sample after purifying in step (2) under the same conditions
Liquid injects GC-MS/MS and is measured, and records the chromatographic peak area of fluorine ether bacterium amide in sample liquid, substitutes into standard curve, obtains
Fluorine ether bacterium amide content in sample liquid, then obtains fluorine ether bacterium amide in sample according to the Mass Calculation of sample representated by sample liquid residual
Allowance.If fluorine ether bacterium amide residual quantity exceedes the range of linearity upper limit in upper machine solution, need to be with constant volume solvent by dilute for upper machine solution concentration
Release to the range of linearity.
Step (1) if in sample dehydrated vegetables and fruit, need to reduce sample weighting amount, and add suitable quantity of water and fully infiltrate.
Add sodium chloride when step (1) uses acetonitrile extraction to saltout, use the acetonitrile solution containing 1% acetic acid to add when extracting
Sodium acetate is saltoutd；A certain amount of water need to be added when the sample of moisture content less is saltoutd.
The dispersive solid-phase extraction agent of step (2) mesostroma is by anhydrous magnesium sulfate, C18Form with PSA, nothing in every volume extracting solution
Water magnesium sulfate, C18It is respectively 150mg, 50mg and 25mg with PSA addition.
In step (4), GC conditions is: chromatographic column: HP-5MS capillary chromatographic column, column length 30m, internal diameter 0.25
Mm, thickness 0.25 μm；Injector temperature 250 DEG C；Carrier gas: He, not shunt mode sample introduction, sample size: 1 μ L；Constant current is entered
Original mold formula, flow velocity 1.2mL/min；Heating schedule: initial temperature 60 DEG C keeps 2min, rises to 200 DEG C with the speed of 20 DEG C per minute,
Then rise to 220 DEG C with the speed of 2 DEG C per minute, then rise to 280 DEG C with the speed of 20 DEG C per minute, keep 10min；Transmission line
Temperature: 290 DEG C.
In step (4), Mass Spectrometry Conditions is: ionization pattern: electron impact ionization (EI, 70eV)；Ion source temperature is 280 DEG C,
Collision gas: methane gas；Multiple-reaction monitoring scan mode SRM, the parent ion of fluorine ether bacterium amide is 379.8～381.2, and daughter ion divides
It is not 205.8～206.2 and 190.7～191.1.
When step (4) measures sample liquid and extraction standard working solution, if sample liquid Pesticides chromatographic peak retention time and standard
In solution, corresponding pesticide retention time is consistent, and in the sample mass spectrum after background correction, selected ion all occurs,
And abundance of ions is more consistent than the abundance of ions ratio with standard solution, then can determine whether sample liquid exists this pesticide；If above-mentioned two
Individual condition can not meet simultaneously, then judge without this kind of pesticide.
The beneficial effects of the present invention is:
The present invention utilizes dispersive solid-phase extraction technology, establishes simplicity, quickly and can be prevented effectively from the sample of sample mesostroma interference
Product pre-treating method, this pre-treating method is combined GC-MS/MS be applied in vegetable and fruit the qualitative confirmation of fluorine ether bacterium amide and
Detection by quantitative, average recovery rate is 89.1%～95.0%, and average relative standard's deviation (RSD) is 4.0%～5.3%, detection limit
Less than 1.02 μ g/kg, there is easy and simple to handle, quick, accurate, highly sensitive and reproducible advantage.0.01mg/kg can be met
" uniform limit " technology requirement of residue limits, for ensureing that our people's food safety, export abroad trade sound development provide
Strong technical support.
Accompanying drawing explanation
Fig. 1 is the selection chromatography of ions figure being added on the 5ng/mL fluorine ether bacterium amide mark liquid in blank Fructus Mali pumilae substrate.
Fig. 2 is the selection chromatography of ions figure of the Fructus Mali pumilae blank sample of not fluorine-containing ether bacterium amide.
Fig. 3 is with the Fructus Mali pumilae blank sample of not fluorine-containing ether bacterium amide as substrate～the fluorine ether bacterium amide standard working curve of preparation.
Detailed description of the invention
Now with following embodiment, the present invention is described, but is not to limit the scope of the present invention.
The instrument used in embodiment and reagent
T18Basic homogenizer (IKA, Germany)；5810R centrifuge (Eppendorf, Germany)；MS3 is basic
Type vortex mixer (IKA, Germany)；Trace 1310 gas chromatogram-TSQ 8000 mass spectrograph (Thermo, USA)；Second
Two amine-n-propyl silane (PSA) adsorbents (40～60 μm), octadecylsilane Bonded Phase (C18) cleanser (40～60
μm) all it is purchased from Anjelen Sci. & Tech. Inc of the U.S..
Reagent: acetonitrile, acetone, normal hexane (HPLC level, Merke, Germany)；Acetic acid (HPLC level, CNW,
Germany)；Anhydrous magnesium sulfate, sodium chloride and sodium acetate are analytical pure, are purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Standard substance: purity 97.8%, purchased from Shandong United Pesticide Industry Co., Ltd..
Embodiment 1: the detection of fluorine ether bacterium amide residual quantity in Fructus Mali pumilae
(1) sample pre-treatments
Weighing through the abundant Fructus Mali pumilae 10.0g mixed in 50mL centrifuge tube, accurately add 20mL acetonitrile, homogenizing extracts 1min, addition 3
G anhydrous magnesium sulfate and 2g sodium chloride, after vortex 1min, 7000r/min is centrifuged 5min.After Li Xin, take 6mL acetonitrile extraction liquid
It is transferred to equipped with 900mg anhydrous magnesium sulfate, 300mg C18With in the centrifuge tube of 150mg PSA, vortex 1min, 5000r/min
Centrifugal 5min.Take 1mL supernatant in nitrogen blowpipe, dry up in 40 DEG C of nitrogen, add acetone that 1mL volume ratio is 1/1/just oneself
Alkane mixed solvent dissolved residue, after vortex mixed film, moves in sample injection bottle and treats that GC-MS/MS measures.
(2) preparation of standard working solution
Accurately weighing 25 ± 0.1mg standard substance in 25mL volumetric flask, dissolve with acetonitrile, constant volume obtains 1000.0 μ g/mL standard reserving solutions；
Pipette 1.0mL standard reserving solution to be placed in 100mL volumetric flask, obtain with the acetone that volume ratio is 1/1/normal hexane mixed solvent constant volume
10.0 μ g/mL standard intermediate liquids.The Fructus Mali pumilae blank sample of not fluorine-containing ether bacterium amide is processed by above-mentioned pre-treatment step, takes 10mL
Blank extraction and cleaning liquid, after being concentrated to dryness, adds acetone/normal hexane mixed solvent vortex that 10mL volume ratio is 1/1 and dissolves, use
This solvent is finally configured to 1,2,5,10,20,50 μ g/L extraction standard working solutions.
(3) gas chromatogram tandem mass spectrometry (GC-MS/MS) measures
The standard working solution of variable concentrations gradient is injected separately into GC-MS/MS, carries out quantitatively dividing of fluorine ether bacterium amide content with external standard method
Analysis, i.e. carries out regression analysis with the chromatographic peak area of standard working solution to its respective concentration, obtains standard curve；At the same terms
Lower sample extracting solution is injected GC-MS/MS it be measured, record the chromatographic peak area of fluorine ether bacterium amide in sample liquid, substitute into mark
Directrix curve, obtains fluorine ether bacterium amide content in sample liquid, then obtains in sample according to the Mass Calculation of sample representated by sample liquid
Fluorine ether bacterium amide residual quantity.
Wherein chromatographic condition is:
Chromatographic column: HP-5MS capillary chromatographic column, column length 30m, internal diameter 0.25mm, thickness 0.25 μm.
Injector temperature: 250.0 DEG C, sample introduction pattern: Splitless injecting samples, sample size: 1 μ L.
Carrier gas: He, constant current mode, flow velocity 1.2mL/min.
Furnace box heating schedule: initial temperature 60 DEG C keeps 2min, rises to 200 DEG C with the speed of 20 DEG C per minute, then with per minute
The speed of 2 DEG C rises to 220 DEG C, then rises to 280 DEG C with the speed of 20 DEG C per minute, keeps 10min.
Transmission line temperature: 290 DEG C.
Wherein, mass spectrometry parameters is:
Ionization pattern: electron impact ionization, i.e. EI pattern, energy 70eV.
Ion source temperature: 280 DEG C.
Collision gas: methane gas.
Scan mode: multiple-reaction monitoring (SRM) scan pattern.
MRM detection parameter is shown in Table 1.
Table 1: the MRM of embodiment 1 detects parameter
*For quota ion pair.
Qualitative Identification: at identical conditions, if sample liquid Pesticides chromatographic peak retention time pesticide corresponding to standard solution
Retention time is consistent, and in the sample mass spectrum after background correction, selected ion all occurs, and abundance of ions
Ratio is consistent with the abundance of ions ratio of standard solution, then can determine whether to exist in sample liquid this pesticide；If above-mentioned two condition can not be same
Time meet, then judge without this kind of pesticide.
With the chromatographic peak area of standard working solution, its respective concentration is carried out regression analysis, obtain standard working curve such as table 1.
The standard curve of fluorine ether bacterium amide in table 1 Fructus Mali pumilae bare substrate
| Title | Retention time (min) | Regression equation | Correlation coefficient |
| Fluorine ether bacterium amide LH-2010A | 17.85 | Y=107919X+30605 | 0.9999 |
Recovery of standard addition and repeatability:
The fluorine ether bacterium amide standard of 10,20 and 200 g/kg3 concentration levels of μ is added in the Fructus Mali pumilae of not fluorine-containing ether bacterium amide
Solution, carries out the determination of residual amount by above-mentioned process step after pesticide adds 30min, and the sample that 200 μ g/kg add concentration is to be measured
Liquid volume ratio be 1/1 acetone/normal hexane mixed solvent dilute 5 times after measure with GC-MS again.Concentration and pesticide will be measured
Theoretical concentration of adding compares, and obtains pesticide TIANZHU XINGNAO Capsul, each pitch-based sphere parallel assay 6 times, obtains its relative standard
Deviation, measurement result is shown in Table 2.As can be seen from Table 2, in 3 mark-on levels, the average recovery rate of fluorine ether bacterium amide is
92.8%～95.0%, average relative standard's deviation (RSD) is 4.3%～5.0%, illustrates that the response rate of the inventive method is higher,
Reproducible.
The response rate of table 2 fluorine ether bacterium amide and repeatability (n=6)
Detection limit:
The fluorine ether bacterium amide substrate standard working solution of variable concentrations is injected GC-MS/MS, molten with least concentration extraction standard
3 times of signal to noise ratios at liquid chromatography peak and the cycles of concentration (cycles of concentration of Fructus Mali pumilae is 0.5 times) of sample handling processes calculate detection limit,
The detection of fluorine ether bacterium amide is limited to 0.192 μ g/kg.
Embodiment 2: the detection of fluorine ether bacterium amide residual quantity in dehydration Capsicum annuum L.
(1) sample pre-treatments
Weigh through the abundant dehydration Capsicum annuum L. 2.0g mixed in 50mL centrifuge tube, after adding 5mL water recovery 30min, accurately add 20
The mL acetonitrile solution containing 1% acetic acid, homogenizing extracts 1min, adds 3g anhydrous magnesium sulfate, 2g sodium acetate and 2mL water, vortex 1
After min, 7000r/min is centrifuged 5min.After Li Xin, take 6mL acetonitrile extraction liquid be transferred to equipped with 900mg anhydrous magnesium sulfate, 300
mg C18With in the centrifuge tube of 150mg PSA, vortex 1min, 5000r/min are centrifuged 5min.Take 1mL supernatant in nitrogen blowpipe
In, dry up in 40 DEG C of nitrogen, add acetone/normal hexane mixed solvent dissolved residue that volume ratio is 1/1, after vortex after mixing, move
Enter and sample injection bottle being treated, GC-MS/MS measures.
(2) preparation of standard working solution
Acetone/normal hexane mixed solvent that 1000ng/mL standard reserving solution volume ratio is 1/1 is diluted to 10ng/mL standard intermediate liquid,
The dehydration Capsicum annuum L. blank sample of not fluorine-containing ether bacterium amide is processed by above-mentioned pre-treatment step, takes 10mL blank extraction and cleaning liquid, dense
Be reduced to dry after, add acetone/normal hexane mixed solvent vortex that 10mL volume ratio is 1/1 and dissolve, with this solvent be finally configured to 1,
2,5,10,20,50 μ g/L extraction standard working solution.
(3) gas chromatogram tandem mass spectrometry (GC-MS/MS) measures
Operating procedure, chromatograph are consistent with the mensuration of fluorine ether bacterium amide in above-mentioned Fructus Mali pumilae sample with Mass Spectrometry Conditions.
Qualitative Identification:
Consistent with the mensuration of fluorine ether bacterium amide in above-mentioned Fructus Mali pumilae sample.
Linear relationship:
With the chromatographic peak area of standard working solution, its respective concentration being carried out regression analysis, obtaining standard working curve is
Y=126125X+14390, correlation coefficient is 0.9998.
Recovery of standard addition and repeatability:
The fluorine ether bacterium amide of 50,100 and 200 3 concentration levels of μ g/kg is added in the dehydration Capsicum annuum L. of not fluorine-containing ether bacterium amide
Standard solution, carries out the determination of residual amount by above-mentioned process step after pesticide adds 30min, mensuration concentration is added with pesticide theory
Add concentration to compare, obtain pesticide TIANZHU XINGNAO Capsul, each pitch-based sphere parallel assay 6 times, obtain its relative standard deviation,
Measurement result is shown in Table 3.As can be seen from Table 3, in 3 mark-on levels, the average recovery rate of fluorine ether bacterium amide be 89.1%～
95.0%, average relative standard's deviation (RSD) is 4.3%～5.0%, illustrates that the response rate of the inventive method is high, reproducible.
The response rate of table 3 fluorine ether bacterium amide and repeatability (n=6)
Detection limit:
The fluorine ether bacterium amide substrate standard working solution of variable concentrations is injected GC-MS/MS, molten with least concentration extraction standard
3 times of signal to noise ratios at liquid chromatography peak and the cycles of concentration (cycles of concentration of dehydration Capsicum annuum L. is 0.1 times) of sample handling processes calculate inspection
Rising limit, the detection of fluorine ether bacterium amide is limited to 1.02 μ g/kg.
Above embodiment is only to be described the preferred embodiment of the present invention, not limits the scope of the present invention
Fixed, on the premise of designing spirit without departing from the present invention, it is each that technical scheme is made by this area ordinary skill technology
Plant modification and improvement, all should fall in the protection domain that claims of the present invention determines.
Claims (4)
1. a GC-MS/MS measures the method for fluorine ether bacterium amide residual in fruit and vegerable, it is characterised in that said method comprising the steps of:
(1) extract
Weigh vegetable and fruit sample in tool plug centrifuge tube, add acetonitrile or the acetonitrile solution homogenizing containing 1% acetic acid extracts 1min, centrifugal after adding the one in sodium chloride or sodium acetate and anhydrous magnesium sulfate vibration；
(2) purify
Pipette sample extracting solution supernatant in centrifuge tube, add Dispersive solid phase extraction agent, vortex oscillation, centrifugal, take after a certain amount of scavenging solution nitrogen dries up, dissolve constant volume with the acetone that volume ratio is 1/1/normal hexane mixed solvent, after crossing film, treat that gas chromatogram tandem mass spectrum (GC-MS/MS) detects；
(3) preparation of standard working solution
The same kind matrix blank sample of not fluorine-containing ether bacterium amide is processed by above-mentioned steps (1), (2), obtain sample extraction and purify residue, vehicle solution is obtained, with the fluorine ether bacterium amide series standard working solution of this solution at least 3 concentration of preparation after adding the mixing of appropriate solvent vortex；
(4) measure and result calculates
In step (4), GC-MS/MS analysis condition is: chromatographic column: HP-5MS capillary chromatographic column, column length 30m, internal diameter 0.25mm, thickness 0.25 μm；Injector temperature 250 DEG C；Carrier gas: He, not shunt mode sample introduction, sample size: 1 μ L；Constant current sample introduction pattern, flow velocity 1.2mL/min；Heating schedule: initial temperature 60 DEG C keeps 2min, rises to 200 DEG C with the speed of 20 DEG C per minute, then rises to 220 DEG C with the speed of 2 DEG C per minute, then rises to 280 DEG C with the speed of 20 DEG C per minute, keeps 10min；Transmission line temperature is 290 DEG C；Ionization pattern: electron impact ionization；Ion source temperature is 280 DEG C, collision gas: argon；Multiple-reaction monitoring scan mode, the parent ion of fluorine ether bacterium amide is 379.8～381.2, and daughter ion is respectively 205.8～206.2 and 190.7～191.1；
The standard working solution of each Concentraton gradient in step (3) is carried out GC-MS/MS mensuration, with the chromatographic peak area of standard working solution, its respective concentration is carried out regression analysis, obtain extraction standard working curve；Sample liquid after purifying in step (2) under the same conditions is injected GC-MS/MS and is measured, record the chromatographic peak area of fluorine ether bacterium amide in sample liquid, substitute into extraction standard working curve, obtain fluorine ether bacterium amide content in sample liquid, then obtain fluorine ether bacterium amide residual quantity in sample according to the Mass Calculation of sample representated by sample liquid；If fluorine ether bacterium amide residual quantity exceedes the range of linearity upper limit in upper machine solution, within upper machine solution concentration need to being diluted to the range of linearity with constant volume solvent.
A kind of GC-MS/MS the most according to claim 1 measures the method for fluorine ether bacterium amide residual in fruit and vegerable, it is characterised in that step (1) if in vegetable and fruit sample dehydrated sample, need to reduce sample weighting amount, and add suitable quantity of water and fully infiltrate.
A kind of GC-MS/MS the most according to claim 1 measures the method for fluorine ether bacterium amide residual in fruit and vegerable, it is characterized in that, sodium chloride need to be added when step (1) uses acetonitrile extraction to saltout, use the acetonitrile solution containing 1% acetic acid need to add sodium acetate when extracting and saltout.
A kind of GC-MS/MS the most according to claim 1 measures the method for fluorine ether bacterium amide residual in fruit and vegerable, it is characterised in that the dispersive solid-phase extraction agent of step (2) mesostroma is by anhydrous magnesium sulfate, C18Form with PSA, anhydrous magnesium sulfate, C in every milliliter of extracting solution18It is respectively 150mg, 50mg and 25mg with PSA addition.
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| CN105548440A (en) * | 2016-01-30 | 2016-05-04 | 崔淑华 | GC (Gas Chromatograph)-EI (Electronic Impact Ionization)-MS (Mass Spectrometry) determining method of residual quantity of penflufen |
| CN105717220A (en) * | 2016-01-30 | 2016-06-29 | 崔淑华 | GC-MS/MS rapid measurement method for penflufen residual amount |
| CN105510506A (en) * | 2016-01-30 | 2016-04-20 | 崔淑华 | Method for GC-MS/MS (gas chromatography tandem mass spectrometry) determination of penflufen remains in fruits and vegetables |
| CN105717212A (en) * | 2016-01-30 | 2016-06-29 | 崔淑华 | GC-MS/MS method for determining fluxapyroxad residues in fruits and vegetables |
| CN105717223A (en) * | 2016-01-30 | 2016-06-29 | 崔淑华 | GC-EI-MS method for determining penflufen residues in fruits and vegetables |
| CN115201353A (en) * | 2022-06-16 | 2022-10-18 | 广州海关技术中心 | Method for detecting residual quantity of kresoxim-methyl in vegetables and fruits |
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