CN105699520A - Quick measurement method for fluxapyroxad residue in animal derived foods - Google Patents
Quick measurement method for fluxapyroxad residue in animal derived foods Download PDFInfo
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- CN105699520A CN105699520A CN201610070241.2A CN201610070241A CN105699520A CN 105699520 A CN105699520 A CN 105699520A CN 201610070241 A CN201610070241 A CN 201610070241A CN 105699520 A CN105699520 A CN 105699520A
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- SXSGXWCSHSVPGB-UHFFFAOYSA-N fluxapyroxad Chemical group FC(F)C1=NN(C)C=C1C(=O)NC1=CC=CC=C1C1=CC(F)=C(F)C(F)=C1 SXSGXWCSHSVPGB-UHFFFAOYSA-N 0.000 title claims abstract description 52
- 235000013305 food Nutrition 0.000 title claims abstract description 26
- 241001465754 Metazoa Species 0.000 title claims abstract description 20
- 238000000691 measurement method Methods 0.000 title abstract 2
- 239000005788 Fluxapyroxad Substances 0.000 claims abstract description 48
- 238000001514 detection method Methods 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 19
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000000605 extraction Methods 0.000 claims abstract description 18
- 239000011159 matrix material Substances 0.000 claims abstract description 6
- 238000000746 purification Methods 0.000 claims abstract description 5
- 239000000523 sample Substances 0.000 claims description 48
- 239000000047 product Substances 0.000 claims description 21
- 239000007788 liquid Substances 0.000 claims description 17
- 239000000243 solution Substances 0.000 claims description 16
- 150000002500 ions Chemical class 0.000 claims description 15
- 239000012224 working solution Substances 0.000 claims description 15
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 14
- 238000000451 chemical ionisation Methods 0.000 claims description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 11
- 239000012086 standard solution Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 238000003556 assay Methods 0.000 claims description 8
- 239000007789 gas Substances 0.000 claims description 8
- 150000002632 lipids Chemical class 0.000 claims description 8
- 238000001819 mass spectrum Methods 0.000 claims description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 7
- 238000002098 selective ion monitoring Methods 0.000 claims description 7
- 239000011780 sodium chloride Substances 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000012496 blank sample Substances 0.000 claims description 5
- 239000012046 mixed solvent Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- 238000004458 analytical method Methods 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 238000000611 regression analysis Methods 0.000 claims description 4
- NDAUXUAQIAJITI-UHFFFAOYSA-N albuterol Chemical compound CC(C)(C)NCC(O)C1=CC=C(O)C(CO)=C1 NDAUXUAQIAJITI-UHFFFAOYSA-N 0.000 claims description 3
- 230000005540 biological transmission Effects 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
- 238000012544 monitoring process Methods 0.000 claims description 3
- 229960002052 salbutamol Drugs 0.000 claims description 3
- 239000000575 pesticide Substances 0.000 abstract description 18
- 238000011084 recovery Methods 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 3
- 239000006185 dispersion Substances 0.000 abstract description 2
- 238000010812 external standard method Methods 0.000 abstract description 2
- 239000012535 impurity Substances 0.000 abstract description 2
- 238000000262 chemical ionisation mass spectrometry Methods 0.000 abstract 1
- 241000282894 Sus scrofa domesticus Species 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 235000013399 edible fruits Nutrition 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 235000013339 cereals Nutrition 0.000 description 4
- 244000144972 livestock Species 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 239000003899 bactericide agent Substances 0.000 description 3
- 235000019625 fat content Nutrition 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 238000004445 quantitative analysis Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 210000000582 semen Anatomy 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 235000013311 vegetables Nutrition 0.000 description 3
- 241000123650 Botrytis cinerea Species 0.000 description 2
- 241000221785 Erysiphales Species 0.000 description 2
- 241001361634 Rhizoctonia Species 0.000 description 2
- 241000813090 Rhizoctonia solani Species 0.000 description 2
- 241001674391 Sphaerulina musiva Species 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 238000003705 background correction Methods 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000000417 fungicide Substances 0.000 description 2
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000004451 qualitative analysis Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 241001465180 Botrytis Species 0.000 description 1
- 235000005637 Brassica campestris Nutrition 0.000 description 1
- 241001301148 Brassica rapa subsp. oleifera Species 0.000 description 1
- 241001157813 Cercospora Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 241000221787 Erysiphe Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- 241000283898 Ovis Species 0.000 description 1
- 241000440444 Phakopsora Species 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 241000221300 Puccinia Species 0.000 description 1
- 239000005869 Pyraclostrobin Substances 0.000 description 1
- 241001533598 Septoria Species 0.000 description 1
- 102000019259 Succinate Dehydrogenase Human genes 0.000 description 1
- 108010012901 Succinate Dehydrogenase Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 150000002430 hydrocarbons Chemical group 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002222 matrix solid-phase dispersion Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000003987 organophosphate pesticide Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- HZRSNVGNWUDEFX-UHFFFAOYSA-N pyraclostrobin Chemical compound COC(=O)N(OC)C1=CC=CC=C1COC1=NN(C=2C=CC(Cl)=CC=2)C=C1 HZRSNVGNWUDEFX-UHFFFAOYSA-N 0.000 description 1
- 239000002728 pyrethroid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000004763 spore germination Effects 0.000 description 1
- 230000028070 sporulation Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 150000003852 triazoles Chemical class 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
-
- 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
-
- 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
- G01N2030/025—Gas chromatography
-
- 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
- G01N2030/062—Preparation extracting sample from raw material
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a quick measurement method for fluxapyroxad residue in animal derived foods. The method is mainly used for measuring the content of residual fluxapyroxad in the animal derived foods with high fat content. The method comprises the following steps: homogeneously extracting residual fluxapyroxad from a sample using acetonitrile; performing EMR (enhanced matrix removal) matrix dispersion purification as well as reverse-extraction tube extraction and concentration on the extract; performing GC-NCI-MS (gas chromatography-negative chemical ionization-mass spectrometry) detection; establishing a corrected standard work curve using a blank matrix solution without the pesticide to be detected; and quantifying by an external standard method. With average recovery rate of 86.7-93.8%, average RSD (relative standard deviation) of 6.0-7.8% and detection limit lower than 2.46mug/kg, the method has the advantages of simplicity and quickness in detection, good impurity removal effect, high sensitivity, strong characteristic feature, good repeatability and qualitative and quantitative accuracies and can meet the technical requirements on the safety detection of corresponding products in countries including America, Europe and Japan and provide powerful technical support to guarantee the food safety for national people and the sound development of export trade.
Description
Technical field
The present invention relates to the rapid assay methods of fluxapyroxad residual in a kind of animal derived food, more specifically adopt Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) qualitative, quantitative to measure the method for the fluxapyroxad content of residual in the animal derived food that the fat contents such as animal muscle and goods such as Carnis Sus domestica, beef, Carnis caprae seu ovis, Carnis Gallus domesticus are high, belong to the determination techniques field of persticide residue。
Background technology
Fluxapyroxad (fluxapyroxad) is the succinate dehydrogenase inhibitors series bactericidal agent developed by BASF AG。Chemical name: 3-difluoromethyl-1-methyl-N-(3', 4', 5'-trifluoro-biphenyl-2-base)-1H-pyrazole-4-carboxamide；Molecular formula: C18H12F5N3O；Relative molecular mass: 381.3；Fusing point: 157 DEG C；Relative density: 1.42。Dissolubility (g/L, 20 DEG C) in solvent: acetone > 250, acetonitrile 168, ethyl acetate 123, methanol 53.4, toluene 20.0。The chemical structural formula of fluxapyroxad is as follows:
Fluxapyroxad has efficiently, wide spectrum, holding effect, excellent Uptake and translocation, have advantages such as preventive and therapeutic action simultaneously。It can suppress the elongation of spore-germination, spore pipe, mycelial growth and Sporulation, can effectively prevent and treat the Major Diseases of corn, Semen sojae atricolor, Semen Maydis, Brassica campestris L and special crop etc.。This product prevents and treats a series of fungal disease by blade face and seed treatment, such as the disease caused by Septoria musiva (Septoria), the pathogen of Botrytis cinerea (Botrytis), powdery mildew (Erysiphe), tail spore bacterium (Cercospora), handle rest fungus (Puccinia), rhizoctonia (Rhizoctonia), nuclear cavity bacteria (Pyrenophoraspp.) etc. on corn, Semen sojae atricolor, fruit tree and vegetable。It is particularly well-suited to bean, gray mold (Botrytiscinerea), rust, powdery mildew and the microbial disease of septoria musiva that preventing and treating is caused by alternaric bacteria (Alternariaspp.), by the microbial disease of rod method etc. on soybean rust (Phakopsora), the disease that Cotton Gossypii is caused by Rhizoctonia solani Kuhn (Rhizoctoniasolani) and Helianthi and oilseeds dish。Under all test doses, all crops are very safe。Fluxapyroxad suitability is strong, can with the composite use of other products of pyraclostrobin, triazole bactericidal agent and BASF。Research shows, fluxapyroxad has better activity than commercially available acid amide fungicides, and no matter this product is active, multi-functional, or is all that modern fungicides establishes new mark post in interior absorption and rain wash resistance etc.。BASF intends to introduce fluxapyroxad more than 50 country in the world, for 100 various crop。2011, fluxapyroxad was first in Britain's registration and listing, and the country of existing registration and listing includes Australia, the U.S., Canada, state of European Union 13, Brazilian and Chinese etc.。Company predict, this product year peak value sales volume up to 600,000,000 Euros。This is in all SDHI series bactericidal agents, the product that expected value that development company expresses is the highest。
Along with the registration of fluxapyroxad, popularization and use, its maximum allowable residual quantity (MRL) in the food agricultural product such as veterinary antibiotics, Cereals and livestock products has been formulated as countries such as the U.S. in China's main exit market, Canada, as the U.S. specifies that fluxapyroxad MRL in fruit and vegerable is 0.5～30mg/kg, in the animal derived food such as livestock products and aquatic products, MRL is 0.01mg/kg, and in nut and Cereals, MRL is 0.06～15mg/kg；Canada regulation fluxapyroxad MRL in fruit and vegerable is 0.02～2mg/kg, and in the animal derived food such as livestock products and aquatic products, MRL is 0.01～0.05mg/kg, and in nut and Cereals, MRL is 0.01～3mg/kg；If European Union, Japan and other countries regulation field use pesticide not register in this country, when not formulating corresponding residue limits standard, it is exported to the food agricultural product of its country and includes residue limits in the animal derived foods such as livestock meat and all carry out " uniform limit " of 0.01mg/L。
Present stage, the research of fluxapyroxad determination of residual amount method is less, the detection method of report is mainly in vegetable and fruit fluxapyroxad method for detecting residue, these detection methods all adopt Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) to measure the detection method of fluxapyroxad residual quantity in vegetable and fruit, use LC-MS/MS to measure food Residual Pesticides in Farm Produce and have quickly, easy, sensitivity advantages of higher, but due to its price costly, a lot of testing agencies, enterprise or scientific research institutions do not configure this instrument or configuration number of units is less, when adopting LC-MS/MS detection due to different compounds, different mobile phases or chromatographic column need to be used, chromatographic column constantly changed by such needs, mobile phase also expends the long time system is balanced, this constrains the application of LC-MS/MS to a certain extent。The gaschromatographic mass spectrometry (GC-NCI-MS) in outfit negative chemical ionization source is analyzed food Residual Pesticides in Farm Produce tool and is had great advantage, Negative chemical ionization (NCI source) is referred to as mass spectrum " soft ionization source ", analyte containing electronegativity group is had high selectivity and high sensitivity, owing to its characteristic is strong, when utilizing it to carry out retention analysis, matrix interference is little, very accurately object can be carried out qualitative and quantitative analysis。Existing various testing agencies and enterprise have all purchased gas chromatograph-mass spectrometer (GC-MS), also generally all it is equipped with Negative chemical ionization (NCI), now a lot of class pesticide all contain electronegativity group, organochlorine and pyrethroid pesticide molecule and mostly contain the strong electronegative group such as-F ,-Cl ,-Br or-COO-；Organophosphorus pesticide molecule mostly contains=S ,-OR ,-P, the electronegativity group such as-O-,-Cl or-P=O；And the novel agrochemical developed in recent years contains-F group mostly, therefore, GC-NCI-MS is used can conveniently to realize the multi-residue analysis of Multiple Pesticides, compared with GC-EI-MS, better capacity of resisting disturbance, less sensitivity and better selectivity can be obtained, but have no the report of the GC-NCI-MS detection method of fluxapyroxad residual quantity in food agricultural product up to now。Additionally, it is known that, QuEChERS pretreatment technology has become sample-pretreating method most widely used in pesticide residue analysis, but QuEChERS method is primarily adapted for use in the High water cut substrate of low-fat content, such as most fruit and vegetable, when in the face of high fat content substrate limited in one's ability, it is generally required to increase normal hexane grease removal, freezing sample extracting solution goes the steps such as fat, but limited efficiency。Some company is proposed enhancement mode lipid and removes patented product-EnhancedMatrixRemoval, EMR-lipid in recent years, is mainly used in removing the lipid chaff interference with straight chain hydrocarbon structure, including free fatty, cholesterol, triglyceride, phospholipid etc.。This product has been made into matrix solid phase dispersion extractant, either directly through dispersive solid-phase extraction flow process, the lipid impurities in extracting solution can be removed after hydroactivated, the present invention applies when this pre-treating method purifies animal derived food and finds, the method is simply efficient, lipid material removal effect is good, sets up the detection method of fluxapyroxad residual quantity in the dispersion purification of EMR substrate, Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) qualitative and quantitative analysis animal derived food significant。
Summary of the invention
It is an object of the invention to provide the rapid assay methods of fluxapyroxad residual in a kind of animal derived food, be mainly used in measuring fluxapyroxad residual quantity in the food agricultural product that the fat contents such as animal derived food are high。
For realizing object above, the technical solution adopted in the present invention is: the rapid assay methods of fluxapyroxad residual in a kind of animal derived food, comprises the steps:
(1) extract
Weigh mixing sample in tool plug centrifuge tube, after adding suitable quantity of water recovery, be quantitatively adding acetonitrile solution homogenizing and extract, be subsequently adding sodium chloride and anhydrous magnesium sulfate, centrifugal after violent vortex 1min。
Purify
Enhancement mode lipid is removed product E MR water activate, pipette sample extracting solution and purify in pipe in the EMR activated, after vortex 1min, 7000r/min is centrifuged 5min, shifts whole supernatant and saltouts to the centrifuge tube equipped with anhydrous magnesium sulfate and sodium chloride, after vortex centrifugal, pipette the supernatant of certain volume, be concentrated into after doing, dissolve constant volume with the acetone that volume ratio is 1/1/normal hexane mixed solvent, after crossing film, treat Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detection。
(3) preparation of standard working solution
The same kind matrix blank sample of not Fluxapyroxad-containsterilization is processed by above-mentioned steps (1), (2), obtain sample extraction and purify residue, adding appropriate solvent and standard solution, vortex mixes, and is configured to the fluxapyroxad series standard working solution of at least 3 concentration。
(4) measure and result calculates
The standard working solution of each Concentraton gradient in step (3) is carried out GC-NCI-MS mensuration, with the chromatographic peak area of standard working solution, its respective concentration is carried out regression analysis, obtain extraction standard working curve；Under the same conditions the sample liquid after purification in step (2) is injected GC-NCI-MS to be measured, record the chromatographic peak area of fluxapyroxad in sample liquid, substitute into extraction standard working curve, obtaining fluxapyroxad content in sample liquid, then the Mass Calculation of sample representated by liquid obtains fluxapyroxad residual quantity in sample per sample；If fluxapyroxad residual quantity exceedes the range of linearity upper limit in upper machine solution, with constant volume solvent, upper machine solution concentration need to be diluted within the range of linearity。
Step (1) if in the sample of the moisture content less such as sample animal livers, suitable quantity of water must be added before extraction and fully infiltrate。
In step (4), GC conditions 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 mode, flow velocity 1.0mL/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: 280 DEG C。
In step (4), Mass Spectrometry Conditions is: ion source temperature 150 DEG C；Quadrupole rod temperature 150 DEG C；Ionization pattern: negative chemical ionization, i.e. NCI pattern, energy 70eV；Scan mode: Salbutamol Selected Ion Monitoring (SIM) pattern, the ion of monitoring is: 361.1,362.1,362.9。
When step (4) measures sample liquid and extraction standard working solution, if sample liquid Pesticides chromatographic peak retention time pesticide retention time corresponding to standard solution 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 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, establish simplicity, quickly and the sample-pretreating method of sample mesostroma interference can be prevented effectively from, this pre-treating method is applied in Cereals, animal derived food the qualitative confirmation of fluxapyroxad and detection by quantitative in conjunction with GC-NCI-MS, average recovery rate is 86.7%～93.8%, average relative standard's deviation (RSD) is 6.0%～7.8%, detection limit, lower than 2.46 μ g/kg, has easy and simple to handle, quick, accurate, highly sensitive and reproducible advantage。The U.S., European Union, the Japan and other countries technology requirement to corresponding product safety detection can be met, for ensureing that our people's food safety and export abroad trade sound development provide strong technical support。
Accompanying drawing explanation
The GC-NCI-MS that Fig. 1 is the fluxapyroxad being added in blank Carnis Sus domestica substrate selects chromatography of ions figure。
The GC-NCI-MS that Fig. 2 is the Carnis Sus domestica blank sample of not Fluxapyroxad-containsterilization selects chromatography of ions figure。
The fluxapyroxad standard working curve that Fig. 3 is is substrate preparation with the Carnis Sus domestica blank sample of not Fluxapyroxad-containsterilization。
Detailed description of the invention
Now with following embodiment, the present invention is described, but is not restriction the scope of the present invention。
The instrument used in embodiment and reagent
T18Basic homogenizer (IKA, Germany)；CR21G III centrifuge (Hitachi, Japan)；MS3 basic model vortex mixer (IKA, Germany)；TurboVapLV pattern product automatic concentration instrument (Caliper, USA)；7890N gas chromatogram-5977C mass spectrograph (Agilent, USA)；Enhancement mode lipid is removed product (EnhancedMatrixRemoval, EMR) and is purchased from Anjelen Sci. & Tech. Inc of the U.S.。
Reagent
Acetonitrile, acetone, normal hexane (HPLC level, Merke, Germany)；Anhydrous magnesium sulfate, sodium chloride are analytical pure, all purchased from Chemical Reagent Co., Ltd., Sinopharm Group。
Standard substance: purity 99.5%, purchased from Dr.Ehrenstorfer company of Germany。
Embodiment 1: the detection of fluxapyroxad residual quantity in Carnis Sus domestica
(1) sample pre-treatments
Extract
Weigh the 5g Carnis Sus domestica sample through fully mixing in 50mL centrifuge tube, after adding 10mL water recovery 30min, accurately add 20mL acetonitrile solution, mechanical shaking extraction 20min, supersound extraction 5min, add 3g anhydrous magnesium sulfate and 2g sodium chloride, after vortex 1min, after the centrifugal 5min of 7000r/min, to be clean。
Purify
Add 4mL water in the purification pipe removing product E MR equipped with enhancement mode lipid, vortex makes it fully activate, pipette 6mL sample extracting solution and purify in pipe in the EMR activated, after vortex 1min, 7000r/min is centrifuged 5min, shift whole supernatant to saltout to the centrifuge tube equipped with anhydrous magnesium sulfate and sodium chloride, after vortex centrifugal, pipette 4mL supernatant, it is concentrated into after doing, dissolve constant volume with the acetone that volume ratio is 1/1/normal hexane mixed solvent, after crossing film, treat Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detection。
(2) preparation of standard working solution
Accurately weighing 25 ± 0.1mg standard substance in 25mL volumetric flask, dissolve with methanol, constant volume obtains 1000.0 μ g/mL standard reserving solutions；Pipette 1.0mL standard reserving solution and be placed in 100mL volumetric flask, obtain 10.0 μ g/mL standard intermediate liquids with the acetone that volume ratio is 1/1/normal hexane mixed solvent constant volume；10 μ g/mL standard solution dilutions are made into 5,2,1,0.5,0.2,0.1 μ g/mL standard solution。The Carnis Sus domestica blank sample of not Fluxapyroxad-containsterilization is processed by above-mentioned pre-treatment step, obtain sample extraction and purify residue, this residue adds acetone/normal hexane mixed solvent and the 100 above-mentioned standard solution of μ L that 900 μ L volume ratios are 1/1, vortex mixes, and is made into 10,20,50,100,200,500 μ g/L extraction standard working solutions。
(3) Gas Chromatography-Negative chemical ionization source-mass spectrography (GC-NCI-MS) measures
The standard working solution of variable concentrations gradient is injected separately into GC-NCI-MS, carries out the quantitative analysis of fluxapyroxad content with external standard method, namely with the chromatographic peak area of standard working solution, its respective concentration is carried out regression analysis, obtain standard working curve；Under the same conditions sample extracting solution is injected GC-NCI-MS to be measured, record the chromatographic peak area of fluxapyroxad in sample liquid, substitute into standard working curve, obtaining fluxapyroxad content in sample liquid, then the Mass Calculation of sample representated by liquid obtains fluxapyroxad residual quantity in sample per sample。
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 1.0mL/min。
Stove case 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: 280 DEG C。
Wherein, mass spectrometry parameters is:
Ionization pattern: negative chemical ionization, i.e. NCI pattern, energy 70eV。
Ion source temperature: 150 DEG C；Quadrupole rod temperature 150 DEG C。
Scan mode: Salbutamol Selected Ion Monitoring (SIM) pattern；The ion of SIM monitoring is: 361.1,362.1,362.9, and quota ion is 361.1。
Qualitative Identification: at identical conditions, if sample liquid Pesticides chromatographic peak retention time pesticide retention time corresponding to standard solution 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 condition can not meet simultaneously, then judge without this kind of pesticide。
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=208.32X-1299.8, and correlation coefficient is 0.9992。
Recovery of standard addition and repeatability:
The Carnis Sus domestica of not Fluxapyroxad-containsterilization adds the fluxapyroxad standard solution of 10,20 and 200 g/kg3 concentration levels of μ, add until pesticide and carry out the determination of residual amount by above-mentioned process step after 30min, mensuration concentration and pesticide theory are added concentration compare, obtain pesticide TIANZHU XINGNAO Capsul, each pitch-based sphere parallel assay 6 times, obtaining its relative standard deviation, measurement result is in Table 1。As can be seen from Table 1, in 3 mark-on levels, the average recovery rate of fluxapyroxad is 86.7%～93.8%, and average relative standard's deviation (RSD) is 6.0%～7.8%, illustrates that the response rate of the inventive method is high, reproducible。
The response rate of table 1 fluxapyroxad and repeatability (n=6)
Detection limit:
The fluxapyroxad extraction standard working solution of variable concentrations is injected GC-NCI-MS, calculating detection limit with the cycles of concentration (cycles of concentration of Carnis Sus domestica is for 1.0 times) of 3 times of signal to noise ratios of least concentration extraction standard solution chromatographic peak and sample handling processes, the detection of fluxapyroxad is limited to 2.46 μ g/kg。
Above embodiments is only that the preferred embodiment of the present invention is described; not the scope of the present invention is defined; under the premise designing spirit without departing from the present invention; various modification that technical scheme is made by this area ordinary skill technology and improvement, all should fall in the protection domain that claims of the present invention are determined。
Claims (3)
1. the rapid assay methods of fluxapyroxad residual in an animal derived food, it is characterised in that said method comprising the steps of:
(1) extract
Weigh mixing sample in tool plug centrifuge tube, after adding suitable quantity of water, after adding the extraction of acetonitrile solution homogenizing, add sodium chloride and anhydrous magnesium sulfate, centrifugal after violent vortex 1min；
(2) purify
Enhancement mode lipid is removed product E MR water activate, pipette sample extracting solution and purify in pipe in the EMR activated, after vortex 1min, 7000r/min is centrifuged 5min, shifts whole supernatant and saltouts to the centrifuge tube equipped with anhydrous magnesium sulfate and sodium chloride, after vortex centrifugal, pipette the supernatant of certain volume, be concentrated into after doing, dissolve constant volume with the acetone that volume ratio is 1/1/normal hexane mixed solvent, after crossing film, treat Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detection；
(3) preparation of standard working solution
The same kind matrix blank sample of not Fluxapyroxad-containsterilization is processed by above-mentioned steps (1), (2), obtain sample extraction and purify residue, adding appropriate solvent and standard solution, vortex mixes, and is configured to the fluxapyroxad series standard working solution of at least 3 concentration；
(4) measure and result calculates
The standard working solution of each Concentraton gradient in step (3) is carried out GC-NCI-MS mensuration, with the chromatographic peak area of standard working solution, its respective concentration is carried out regression analysis, obtain extraction standard working curve；Under the same conditions the sample liquid after purification in step (2) is injected GC-NCI-MS to be measured, record the chromatographic peak area of fluxapyroxad in sample liquid, substitute into extraction standard working curve, obtaining fluxapyroxad content in sample liquid, then the Mass Calculation of sample representated by liquid obtains fluxapyroxad residual quantity in sample per sample；If fluxapyroxad residual quantity exceedes the range of linearity upper limit in upper machine solution, with constant volume solvent, upper machine solution concentration need to be diluted within the range of linearity。
2. the rapid assay methods of fluxapyroxad residual in a kind of animal derived food according to claim 1, it is characterised in that step (1) if in the sample of sample moisture content less, suitable quantity of water must be added before extraction and fully infiltrate。
3. the rapid assay methods of fluxapyroxad residual in a kind of animal derived food according to claim 1, it is characterized in that, in step (4), GC-NCI-MS analysis condition is: chromatographic column: HP-5MS capillary chromatographic column, column length 30m, internal diameter 0.25mm, thickness 0.25 μm；Injector temperature 250.0 DEG C；Carrier gas: He, not shunt mode sample introduction, sample size: 1 μ L；Constant current mode, flow velocity 1.0mL/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: 280 DEG C；Ionization pattern: negative chemical ionization, i.e. NCI pattern, energy 70eV；Ion source temperature 150 DEG C；Scan mode: Salbutamol Selected Ion Monitoring (SIM) pattern, the ion of monitoring is: 361.1,362.1,362.9。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112739441A (en) * | 2018-09-21 | 2021-04-30 | 沃特世科技公司 | Systems and methods for lipid quantification |
| CN116973468A (en) * | 2023-05-06 | 2023-10-31 | 广州海关技术中心 | Method for determining residual quantity of succinate dehydrogenase inhibitor bactericides in food |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104502484A (en) * | 2014-12-30 | 2015-04-08 | 崔淑华 | GC-NCI-MS (Gas Chromatography-Negative Chemical Ionization-Mass Spectrum) detecting method for residual quantity of cyantraniliprole in fruits and vegetables |
| CN104569271A (en) * | 2015-01-09 | 2015-04-29 | 韩超 | Solid-phase extraction-gas chromatography tandem mass spectrometry detection method for pyrazol bactericides in wine |
| CN104597189A (en) * | 2015-01-29 | 2015-05-06 | 山东出入境检验检疫局检验检疫技术中心 | Determining method of Pyrifluquinazon residual quantity |
| CN104655783A (en) * | 2015-03-12 | 2015-05-27 | 郭庆龙 | GC-NCI-MS (gas chromatography-negative chemical ionization-mass spectrometry) determination method of residual amount of metrafenone in fruits and vegetables |
| CN104655763A (en) * | 2015-03-12 | 2015-05-27 | 崔淑华 | GC-NCI-MS (gas chromatography-negative chemical ionization-mass spectrometry) determination method of residual amount of fluoride ether bacterium amide |
-
2016
- 2016-01-30 CN CN201610070241.2A patent/CN105699520A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104502484A (en) * | 2014-12-30 | 2015-04-08 | 崔淑华 | GC-NCI-MS (Gas Chromatography-Negative Chemical Ionization-Mass Spectrum) detecting method for residual quantity of cyantraniliprole in fruits and vegetables |
| CN104569271A (en) * | 2015-01-09 | 2015-04-29 | 韩超 | Solid-phase extraction-gas chromatography tandem mass spectrometry detection method for pyrazol bactericides in wine |
| CN104597189A (en) * | 2015-01-29 | 2015-05-06 | 山东出入境检验检疫局检验检疫技术中心 | Determining method of Pyrifluquinazon residual quantity |
| CN104655783A (en) * | 2015-03-12 | 2015-05-27 | 郭庆龙 | GC-NCI-MS (gas chromatography-negative chemical ionization-mass spectrometry) determination method of residual amount of metrafenone in fruits and vegetables |
| CN104655763A (en) * | 2015-03-12 | 2015-05-27 | 崔淑华 | GC-NCI-MS (gas chromatography-negative chemical ionization-mass spectrometry) determination method of residual amount of fluoride ether bacterium amide |
Non-Patent Citations (3)
| Title |
|---|
| FENGSHOU DONG ET AL: "Simultaneous determination of five pyrazole fungicides in cereals, vegetables and fruits using liquid chromatography/tandem mass spectrometry", 《JOURNAL OF CHROMATOGRAPHY A》 * |
| LIMIAN ZHAO 等: "使用 Agilent Bond Elut EMR-Lipid 增强型脂质去除产品对牛油果中的农药多残留分析进行GC/MS/MS检测", 《HTTP://CN.AGILENT.COM/CS/LIBRARY/APPLICATIONS/5991-6097CHCN.PDF》 * |
| 刘磊 等: "固相萃取-高效液相色谱法测定草莓中氟唑菌酰胺残留量", 《农药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112739441A (en) * | 2018-09-21 | 2021-04-30 | 沃特世科技公司 | Systems and methods for lipid quantification |
| CN116973468A (en) * | 2023-05-06 | 2023-10-31 | 广州海关技术中心 | Method for determining residual quantity of succinate dehydrogenase inhibitor bactericides in food |
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