CN104459001A - Method for measuring residual quantity of chlorantraniliprole in fruit and vegetable - Google Patents
Method for measuring residual quantity of chlorantraniliprole in fruit and vegetable Download PDFInfo
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- CN104459001A CN104459001A CN201410843724.2A CN201410843724A CN104459001A CN 104459001 A CN104459001 A CN 104459001A CN 201410843724 A CN201410843724 A CN 201410843724A CN 104459001 A CN104459001 A CN 104459001A
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- PSOVNZZNOMJUBI-UHFFFAOYSA-N chlorantraniliprole Chemical compound CNC(=O)C1=CC(Cl)=CC(C)=C1NC(=O)C1=CC(Br)=NN1C1=NC=CC=C1Cl PSOVNZZNOMJUBI-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 238000000034 method Methods 0.000 title claims abstract description 18
- 239000005886 Chlorantraniliprole Substances 0.000 title claims abstract description 16
- 235000012055 fruits and vegetables Nutrition 0.000 title claims abstract description 12
- 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
- 238000000605 extraction Methods 0.000 claims abstract description 14
- 238000000746 purification Methods 0.000 claims abstract description 9
- 239000011159 matrix material Substances 0.000 claims abstract description 6
- 239000000523 sample Substances 0.000 claims description 56
- 239000007788 liquid Substances 0.000 claims description 22
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 20
- 239000000243 solution Substances 0.000 claims description 19
- 239000012224 working solution Substances 0.000 claims description 17
- 150000002500 ions Chemical class 0.000 claims description 16
- 239000012086 standard solution Substances 0.000 claims description 16
- 238000000451 chemical ionisation Methods 0.000 claims description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 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 11
- 239000000284 extract Substances 0.000 claims description 10
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 10
- 235000013311 vegetables Nutrition 0.000 claims description 10
- 241000238631 Hexapoda Species 0.000 claims description 9
- 238000004458 analytical method Methods 0.000 claims description 9
- 238000003556 assay Methods 0.000 claims description 9
- 235000013399 edible fruits Nutrition 0.000 claims description 9
- 239000007789 gas Substances 0.000 claims description 9
- 239000012046 mixed solvent Substances 0.000 claims description 9
- JMANVNJQNLATNU-UHFFFAOYSA-N oxalonitrile Chemical compound N#CC#N JMANVNJQNLATNU-UHFFFAOYSA-N 0.000 claims description 8
- 230000008569 process Effects 0.000 claims description 7
- 238000002098 selective ion monitoring Methods 0.000 claims description 7
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 239000012496 blank sample Substances 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 239000001632 sodium acetate Substances 0.000 claims description 6
- 235000017281 sodium acetate Nutrition 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 238000001819 mass spectrum Methods 0.000 claims description 5
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- 150000001408 amides Chemical class 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 239000006228 supernatant Substances 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
- 238000012544 monitoring process Methods 0.000 claims description 3
- 229960002052 salbutamol Drugs 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 230000010355 oscillation Effects 0.000 claims description 2
- 230000002000 scavenging effect Effects 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 17
- 239000000575 pesticide Substances 0.000 abstract description 13
- 238000011084 recovery Methods 0.000 abstract description 10
- 235000013305 food Nutrition 0.000 abstract description 7
- 230000008901 benefit Effects 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 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
- 238000000262 chemical ionisation mass spectrometry Methods 0.000 abstract 1
- 238000007689 inspection Methods 0.000 abstract 1
- 238000011002 quantification Methods 0.000 abstract 1
- 239000003905 agrochemical Substances 0.000 description 14
- 238000004949 mass spectrometry Methods 0.000 description 7
- 244000046052 Phaseolus vulgaris Species 0.000 description 6
- 230000014759 maintenance of location Effects 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 235000005489 dwarf bean Nutrition 0.000 description 5
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 241000607479 Yersinia pestis Species 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000002203 pretreatment Methods 0.000 description 4
- 244000291564 Allium cepa Species 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 235000002732 Allium cepa var. cepa Nutrition 0.000 description 2
- 235000016623 Fragaria vesca Nutrition 0.000 description 2
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 2
- 241000255777 Lepidoptera Species 0.000 description 2
- 231100000703 Maximum Residue Limit Toxicity 0.000 description 2
- 241000256259 Noctuidae Species 0.000 description 2
- 238000003705 background correction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000005516 engineering process Methods 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
- 230000000749 insecticidal effect Effects 0.000 description 2
- 230000002045 lasting effect Effects 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000004451 qualitative analysis Methods 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 241000590412 Agromyzidae Species 0.000 description 1
- 240000006108 Allium ampeloprasum Species 0.000 description 1
- 235000005254 Allium ampeloprasum Nutrition 0.000 description 1
- 235000010167 Allium cepa var aggregatum Nutrition 0.000 description 1
- 235000017060 Arachis glabrata Nutrition 0.000 description 1
- 244000105624 Arachis hypogaea Species 0.000 description 1
- 235000010777 Arachis hypogaea Nutrition 0.000 description 1
- 235000018262 Arachis monticola Nutrition 0.000 description 1
- 241000254127 Bemisia tabaci Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 241001347514 Carposinidae Species 0.000 description 1
- 241001124134 Chrysomelidae Species 0.000 description 1
- 241000254173 Coleoptera Species 0.000 description 1
- 244000307700 Fragaria vesca Species 0.000 description 1
- 240000009088 Fragaria x ananassa Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 244000289433 Pistacia texana Species 0.000 description 1
- 235000004292 Pistacia texana Nutrition 0.000 description 1
- 241000500437 Plutella xylostella Species 0.000 description 1
- 241000255893 Pyralidae Species 0.000 description 1
- 241001253920 Ryania Species 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 244000037666 field crops Species 0.000 description 1
- 235000021393 food security Nutrition 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- ZHNUHDYFZUAESO-UHFFFAOYSA-N formamide Substances NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- -1 methyl-carbamoyl Chemical group 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
- 235000020232 peanut Nutrition 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000002728 pyrethroid Substances 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 150000003335 secondary amines Chemical class 0.000 description 1
- 235000002639 sodium chloride Nutrition 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a method for measuring the residual quantity of chlorantraniliprole in fruit and vegetable. The method comprises performing homogeneous extraction on chlorantraniliprole residue in a sample through acetonitrile or an acetonitrile solution containing 1% of acetic acid; performing dispersed purification on extracted solution through ethylenediamine-N-propyl silicane (PSA) and octadecyl silane bonded phase (C18) matrix; then performing gas chromatograph-negative chemical ionization-mass spectrometry (GC-NCI-MS) detection, establishing a calibration standard curve through blank matrix solution not containing pesticide to be measured, and performing quantitation through the external standard method. The method for measuring the residual quantity of the chlorantraniliprole in fruit and vegetable achieves an average recovery rate of 90.7-101.9%, an average relative standard deviation (RSD) of 5.4-9.6% and a detection limit lower than 0.77 mu g/kg, has the advantages of being convenient and rapid to operate, high in sensitivity and repeatability and accurate in quantitation and quantification, can meet the technical requirements of the corresponding product safety inspection of nations such as the U.S., Japan, the European Union and Canada, and provides powerful technical support for food safety of people and healthy development of export trade in China.
Description
Technical field
The present invention relates to the assay method of Rynaxypyr residual quantity in a kind of fruits and vegetables, be more particularly the method adopting Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) qualitative, quantitative to measure Rynaxypyr content residual in vegetables and fruit, belong to the determination techniques field of persticide residue.
Background technology
Rynaxypyr (Chlorantraniliprole), trade name health is wide, it is the novel O-formammidotiazol-benzamide pesticide of E.I.Du Pont Company's exploitation, Rynaxypyr high-efficiency broad spectrum, good control effects is all had to lepidopterous Noctuidae, Pyralidae, Carposinidae, tortricid, miller section, diamond-back moth section, Gelechidae, Gracilariidae etc., coleoptera Culculionidae can also be controlled, Chrysomelidae; Diptera Agromyzidae; The multiple non-lepidoptera pest such as Bemisia tabaci.Chemical name is: [4-chloro-2-methyl-6-[(methyl-carbamoyl) benzene]-1-(3-chloropyridine-2-base)-1H-pyrazoles-5-formamide, English language Chemical name is called 3-Bromo-N-[4-chloro-2-methyl-6-[(methylamino) carbonyl] phenyl]-1-(3-chloro-2-pyridinyl)-1H-pyrazole-5-carboxamide to the bromo-N-of 3-.CAS accession number is 500008-45-7, and molecular weight is 483.15, and structural formula is:
The a lot of country of Rynaxypyr obtains registration, has achieved all certificates of agriculture chemical registration sales applications in China, can large scale application.Chemical constitution due to Rynaxypyr has the brand-new desinsection principle that other any pesticides do not possess, efficiently can activate insect ryania (muscle) acceptor, calcium ion in excessive release cells in calcium storehouse, insect is caused to be paralysed dead, high to the larva activity of lepidoptera pest, insecticidal spectrum is wide, and lasting effect is good.10-100 is exceeded doubly according to the specific activity other products of current test findings to target pest. and some lepidopterous insects mating process can be caused disorderly, research proves that it can reduce the spawning rate of multiple noctuidae pests, the biological characteristics of and resistance of rainwater washing against good due to its lasting effect, these characteristics are actually perviousness, conduction, chemical stability, high insecticidal activity and cause insect to stop the comprehensive embodiment of effects such as taking food immediately.Therefore determine its than at present most other pesticide have longer and more stable and to crop protective effect.To the biting mouth parts insect in harm field crops, fruit tree, other special crops of vegetables and turf, can provide long-acting, the preventive and therapeutic effect of wide spectrum.
Along with the registration of Rynaxypyr, popularization and use, as U.S. in China's veterinary antibiotics main exit market, European Union, Canada and Japan and other countries, residue limits standard is formulated to it.On October 19th, 2009, the U.S. sends No. G/SPS/N/USA/1935 circular, and Environmental Protection Agency formulates Final Rule to pesticide Rynaxypyr (Chlorantraniliprole) license limitation.Content comprises: the residue limits of Rynaxypyr on Pericarppium Armeniacae Amarum is 5.0ppm; Residue limits on American pistachios is: 0.04ppm; Residue limits on spring onion is: 0.20ppm; Residue limits on strawberry is: 1.2ppm etc.On May 4th, 2010, the U.S. issues circular, intends extending pesticide Rynaxypyr (Chlorantraniliprole) and permits limitation in limited time.What this Final Rule prolongation pesticide Rynaxypyr (Chlorantraniliprole) remained indirectly or unintentionally permits limitation in limited time: Cereals 16 groups of forages, feed, culms; Leek; Green onion; Shallot; Peanut hay; Verdant; Bean feed; Soybean hay and 2 groups of leaf class rhizomes, tuberous vegetable: 0.20ppm.Canada to the maximum residue limit of Rynaxypyr (Chlorantraniliprole) is:
European Union and Japan have also formulated the maximum residue limit of Rynaxypyr in a lot of food agricultural product, to the food agricultural product not formulating maximum permission residue limits, all carry out " uniform limit " of 0.01mg/L.
In recent years, a lot of to the research of Rynaxypyr residues detection method in varieties of food items agricultural product, the detection method of report all adopts liquid chromatography (LC) or Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) to measure the detection method of cyanogen insect amide residual quantity in vegetables and fruit, using LC-MS/MS to measure food Residual Pesticides in Farm Produce has fast, easy, sensitivity advantages of higher, but due to its price costly, a lot of testing agency, enterprise or scientific research institutions do not configure this instrument or configuration number of units is less, during due to different compounds employing LC-MS/MS detection, different mobile phases or chromatographic column need be used, such needs constantly change chromatographic column, mobile phase also expends the long time and balances system, 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 called as mass spectrum " soft ionization source ", to the analysis thing containing electronegativity group, there is high selectivity and high sensitivity, because its characteristic is strong, when utilizing it to carry out retention analysis, matrix interference is little, can carry out qualitative and quantitative analysis very accurately to object.Existing various testing agency and enterprise have all purchased gas chromatograph-mass spectrometer (GCMS) (GC-MS), generally also be provided with Negative chemical ionization (NCI), now a lot of class agricultural chemicals is all containing electronegativity group, and organochlorine and pyrethroid pesticide molecule are mostly containing strong electronegative group such as-F ,-Cl ,-Br or-COO-, organophosphorus pesticide molecule is mostly containing the=electronegativity group such as S ,-OR ,-P ,-O-,-Cl or-P=O, and mostly containing-F group in the novel agrochemical developed in recent years, therefore, use GC-NCI-MS conveniently can realize the multi-residue analysis of Multiple Pesticides, compared with GC-NCI-MS, better antijamming capability can be obtained, lower sensitivity and better selectivity, Rynaxypyr belongs to electronegativity compound, but have no the report of the GC-NCI-MS detection method of Rynaxypyr residual quantity in vegetables and fruit up to now, therefore, the detection method setting up Rynaxypyr residual quantity in Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) qualitative and quantitative analysis vegetables and fruit is significant.
Summary of the invention
The object of this invention is to provide the assay method of Rynaxypyr residual quantity in a kind of fruits and vegetables.
For realizing above object, the technical solution adopted in the present invention is: the assay method of Rynaxypyr residual quantity in a kind of fruits and vegetables, comprises the steps:
(1) extract
Take sample in tool plug centrifuge tube, add acetonitrile or extract 1min containing the acetonitrile solution homogeneous of 1% acetic acid, add the one in sodium chloride or sodium acetate 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 after a certain amount of scavenging solution nitrogen dries up, dissolve constant volume with acetone/normal hexane mixed solvent that volume ratio is 1/1, after crossing film, treat that Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detects.
(3) preparation of standard working solution
During by the same kind matrix blank sample of not containing chlorantraniliprole by above-mentioned steps (1), (2) process, obtain sample extraction purification residue, add appropriate solvent and mixed standard solution, vortex mixes, and is mixed with the Rynaxypyr series hybrid standard working fluid of at least 3 concentration.
(4) Gas Chromatography-Negative chemical ionization source-mass spectroscopy (GC-NCI-MS) measures
The standard working solution of each concentration gradient in step (3) is carried out GC-NCI-MS mensuration, with the chromatographic peak area of standard working solution, regretional analysis is carried out to its respective concentration, obtain standard working curve; Under the same conditions the sample liquid after purification in step (2) is injected GC-NCI-MS to measure, record the chromatographic peak area of Rynaxypyr in sample liquid, substitute into typical curve, obtain Rynaxypyr content in sample liquid, then the Mass Calculation of sample representated by liquid obtains Rynaxypyr residual quantity in sample per sample.If cyanogen insect amide residual quantity exceedes the range of linearity upper limit in upper machine solution, with constant volume solvent, upper machine solution concentration need be diluted within the range of linearity.
Step (1) if in sample dehydrated vegetables and fruit, need sample weighting amount be reduced, and add suitable quantity of water and fully infiltrate.
Add sodium chloride when adopting acetonitrile to extract in step (1) to saltout, add sodium acetate when adopting the acetonitrile solution containing 1% acetic acid to extract and saltout; A certain amount of water need be added when the sample of moisture content less is saltoutd.
The agent of step (2) mesostroma dispersive solid-phase extraction is by anhydrous magnesium sulfate, C
18with PSA composition, anhydrous magnesium sulfate, C in every volume extract
18150mg, 50mg and 25mg is respectively with PSA addition.
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, then rises 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 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: 278,279,280.
When measuring sample liquid and extraction standard working solution in step (4), if sample liquid Pesticides chromatographic peak retention time agricultural chemicals retention time corresponding to standard solution is consistent, and in the sample mass spectrogram after background correction, selected ion all occurs, and abundance of ions than with the abundance of ions of standard solution than consistent, then can judge to exist in sample liquid this agricultural chemicals; If above-mentioned two conditions can not meet simultaneously, then judge not containing this kind of agricultural chemicals.
Beneficial effect of the present invention is:
The present invention utilizes dispersive solid-phase extraction technology, establish sample-pretreating method that is easy, that also can effectively avoid sample mesostroma to disturb fast, this pre-treating method is applied to the qualitative confirmation of Rynaxypyr and quantitatively detection in vegetables and fruit in conjunction with GC-NCI-MS, average recovery rate is 90.7% ~ 101.9%, average relative standard's deviation (RSD) is 5.4% ~ 9.6%, detection limit, lower than 0.77 μ g/kg, has easy and simple to handle, quick, accurate, highly sensitive and reproducible advantage.The technical requirement of the countries such as the U.S., Japan, European Union, Canada to corresponding product safety detection being met, providing strong technical support by for ensureing that our people's food security and export abroad trade develop in a healthy way.
Accompanying drawing explanation
The total chromatogram of GC-NCI-MS Selective ion mode of Fig. 1 to be concentration be Rynaxypyr mark liquid of 100ng/mL.
Fig. 2 is the total chromatogram of GC-NCI-MS Selective ion mode of the apple blank sample of not containing chlorantraniliprole.
Fig. 3 is the total chromatogram of GC-NCI-MS Selective ion mode of the Rynaxypyr be added in blank apple matrix.
The Rynaxypyr standard working curve that Fig. 4 is is substrate preparation with the apple blank sample of not containing chlorantraniliprole.
Embodiment
Now with following embodiment, the present invention is described, but is not limit the scope of the invention.
The instrument used in embodiment and reagent
T18Basic homogenizer (IKA, Germany); 5810R hydro-extractor (Eppendorf, Germany); MS3 basic model vortex mixer (IKA, Germany); 7890N gas chromatography-5975C mass spectrometer (Agilent, USA); Primary secondary amine (PSA) adsorbent (40 ~ 60 μm), octadecylsilane Bonded Phase (C
18) cleanser (40 ~ 60 μm) is all 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 pure for analyzing, all purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Standard substance: purity >=98.0%, purchased from German Dr.Ehrenstorfer company.
Embodiment 1: the detection of Rynaxypyr residual quantity in apple
(1) sample pre-treatments
Taking apple 10.0g through fully mixing in 50mL centrifuge tube, accurately adding 20mL acetonitrile, homogeneous extracts 1min, adds 3g anhydrous magnesium sulfate and 2g sodium chloride, after vortex 1min, and the centrifugal 5min of 7000r/min.After centrifugal, get 6mL acetonitrile extract and be transferred to 900mg anhydrous magnesium sulfate, 300mg C are housed
18with in the centrifuge tube of 150mg PSA, the centrifugal 5min of vortex 1min, 5000r/min.Get 4mL supernatant in nitrogen blowpipe, dry up in 40 DEG C of nitrogen, add acetone/normal hexane mixed solvent dissolved residue that volume ratio is 1/1, after vortex mixed film, move in sample injection bottle and treat that GC-NCI-MS measures.
(2) preparation of standard working solution
Accurately take 25 ± 0.1mg standard items in 25mL volumetric flask, dissolve with acetonitrile, 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/normal hexane mixed solvent constant volume by volume ratio being 1/1; 10 μ g/mL standard solution dilutions are made into 5,2,1,0.5,0.2,0.1 μ g/mL standard solution.Strawberry blank sample not containing spiral shell worm ethyl ester is pressed above-mentioned pre-treatment step process, obtain sample extraction purification residue, acetone/normal hexane mixed solvent and the above-mentioned mixed standard solution of 100 μ L that 900 μ L volume ratios are 1/1 is added in this residue, 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 spectroscopy (GC-NCI-MS) measures
The standard working solution of variable concentrations gradient is injected GC-NCI-MS respectively, carries out the quantitative test of Rynaxypyr content with external standard method, namely with the chromatographic peak area of standard working solution, regretional analysis is carried out to its respective concentration, obtain typical curve; Under the same conditions sample extracting solution is injected GC-NCI-MS to measure, record the chromatographic peak area of Rynaxypyr in sample liquid, substitute into typical curve, obtain Rynaxypyr content in sample liquid, then the Mass Calculation of sample representated by liquid obtains Rynaxypyr residual quantity in sample per sample.If cyanogen insect amide residual quantity exceedes the range of linearity upper limit in upper machine solution, with constant volume solvent, upper machine solution concentration need be diluted within the range of linearity.
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, then rises 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 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: 278,279,280, and quota ion is 278.
Qualitative Identification: at identical conditions, if sample liquid Pesticides chromatographic peak retention time agricultural chemicals retention time corresponding to standard solution is consistent, and in the sample mass spectrogram after background correction, selected ion all occurs, and abundance of ions than with the abundance of ions of standard solution than consistent, then can judge to exist in sample liquid this agricultural chemicals; If above-mentioned two conditions can not meet simultaneously, then judge not containing this kind of agricultural chemicals.
With the chromatographic peak area of standard working solution, regretional analysis is carried out to its respective concentration, obtain standard working curve as table 1.
The typical curve of Rynaxypyr in table 1 apple bare substrate
| Title | Retention time (min) | Regression equation | Related coefficient |
| Rynaxypyr Chlorantraniliprole | 23.39 | Y＝40.12X-202.37 | 0.9996 |
Recovery of standard addition and repeatability:
The Rynaxypyr standard solution of 10,20, a 400 and 1200 μ g/kg4 concentration level is added in the apple of not containing chlorantraniliprole, add after 30min until agricultural chemicals and carry out the determination of residual amount by above-mentioned treatment step, purification constant volume liquid is diluted to residual quantity within the range of linearity with acetone/normal hexane mixed solvent that volume ratio is 1/1 before adding the sample liquid GC-MS mensuration to be measured of concentration by 400 μ g/kg and 1200 μ g/kg.Mensuration concentration and agricultural chemicals theory are added concentration compare, obtain agricultural chemicals TIANZHU XINGNAO Capsul, each Pitch-based sphere replicate determination 6 times, obtain its relative standard deviation, measurement result is in table 2.As can be seen from Table 2, in 4 mark-on levels, the average recovery rate of Rynaxypyr is 90.7% ~ 101.9%, and average relative standard's deviation (RSD) is 5.7% ~ 7.8%, illustrates that the recovery of the inventive method is higher, reproducible.
The recovery of table 2 Rynaxypyr and repeatability (n=6)
Detection limit:
The Rynaxypyr extraction standard working solution of variable concentrations is injected GC-NCI-MS, calculate detection limit with the cycles of concentration (cycles of concentration of apple is 2.0 times) of 3 times of signal to noise ratio (S/N ratio)s of least concentration extraction standard solution chromatographic peak and sample handling processes, detecting of Rynaxypyr is limited to 0.77 μ g/kg.
Embodiment 2: the detection of Rynaxypyr residual quantity in French bean
(1) sample pre-treatments
Take French bean 10.0g through fully mixing in 50mL centrifuge tube, accurately add the acetonitrile solution of 20mL containing 1% acetic acid, homogeneous extracts 1min, adds 3g anhydrous magnesium sulfate and 2g sodium acetate, after vortex 1min, and the centrifugal 5min of 7000r/min.After centrifugal, get 6mL acetonitrile extract and be transferred to 900mg anhydrous magnesium sulfate, 300mg C are housed
18with in the centrifuge tube of 150mg PSA, the centrifugal 5min of vortex 1min, 5000r/min.Get 4mL supernatant in nitrogen blowpipe, 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 in sample injection bottle and treat that GC-NCI-MS measures.
(2) preparation of standard working solution
10 μ g/mL standard solution dilutions are made into 5,2,1,0.5,0.2,0.1 μ g/mL standard solution.By the French bean blank sample of not containing chlorantraniliprole by above-mentioned pre-treatment step process, obtain sample extraction purification residue, acetone/normal hexane mixed solvent and the above-mentioned mixed standard solution of 100 μ L that 900 μ L volume ratios are 1/1 is added in this residue, 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 spectroscopy (GC-NCI-MS) measures
Operation steps, chromatogram are consistent with the mensuration of Rynaxypyr in above-mentioned apple sample with Mass Spectrometry Conditions.
Qualitative Identification:
Consistent with the mensuration of Rynaxypyr in above-mentioned apple sample.
Linear relationship:
Carry out regretional analysis with the chromatographic peak area of standard working solution to its respective concentration, obtaining standard working curve is Y=50.8X-306.68, and related coefficient is 0.9995.
Recovery of standard addition and repeatability:
The Rynaxypyr standard solution of 10,20,200 μ g/kg and 2mg/kg, 4 concentration levels is added in the French bean of not containing chlorantraniliprole, add after 30min until agricultural chemicals and carry out the determination of residual amount by above-mentioned treatment step, purification constant volume liquid is diluted to residual quantity within the range of linearity with acetone/normal hexane mixed solvent that volume ratio is 1/1 before adding the sample liquid GC-MS mensuration to be measured of concentration by 2mg/kg.Mensuration concentration and agricultural chemicals theory are added concentration compare, obtain agricultural chemicals TIANZHU XINGNAO Capsul, each Pitch-based sphere replicate determination 6 times, obtain its relative standard deviation, measurement result is in table 3.As can be seen from Table 3, in 4 mark-on levels, the average recovery rate of Rynaxypyr is 94.4% ~ 100.5%, and average relative standard's deviation (RSD) is 5.4% ~ 9.6%, illustrates that the recovery of the inventive method is high, reproducible.
The recovery of table 3 Rynaxypyr and repeatability (n=6)
Detection limit:
The Rynaxypyr extraction standard working solution of variable concentrations is injected GC-NCI-MS, calculate detection limit with the cycles of concentration (cycles of concentration of French bean is 2.0 times) of 3 times of signal to noise ratio (S/N ratio)s of least concentration extraction standard solution chromatographic peak and sample handling processes, detecting of Rynaxypyr is limited to 0.49 μ g/kg.
Above embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various modification that the common engineering in this area is made technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determine.
Claims (5)
1. the assay method of Rynaxypyr residual quantity in fruits and vegetables, is characterized in that, said method comprising the steps of:
(1) extract
Take vegetables and fruit sample in tool plug centrifuge tube, add acetonitrile or extract 1min containing the acetonitrile solution homogeneous of 1% acetic acid, centrifugal after adding 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, get after a certain amount of scavenging solution nitrogen dries up, dissolve constant volume with acetone/normal hexane mixed solvent that volume ratio is 1/1, after crossing film, treat that Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detects;
(3) preparation of standard working solution
By the same kind matrix blank sample of not containing chlorantraniliprole by above-mentioned steps (1), (2) process, obtain sample extraction purification residue, add appropriate solvent and mixed standard solution, vortex mixes, and is mixed with the Rynaxypyr series hybrid standard working fluid of at least 3 concentration;
(4) mensuration and result calculate
The standard working solution of each concentration gradient in step (3) is carried out GC-NCI-MS mensuration, with the chromatographic peak area of standard working solution, regretional analysis is carried out to its respective concentration, obtain extraction standard working curve; Under the same conditions the sample liquid after purification in step (2) is injected GC-NCI-MS to measure, record the chromatographic peak area of Rynaxypyr in sample liquid, substitute into extraction standard curve, obtain Rynaxypyr content in sample liquid, then the Mass Calculation of sample representated by liquid obtains Rynaxypyr residual quantity in sample per sample.If cyanogen insect amide residual quantity exceedes the range of linearity upper limit in upper machine solution, with constant volume solvent, upper machine solution concentration need be diluted within the range of linearity.
2. the assay method of Rynaxypyr residual quantity in a kind of fruits and vegetables according to claim 1, is characterized in that, step (1) if in vegetables and fruit sample dehydrated sample, need sample weighting amount be reduced, and add suitable quantity of water and fully infiltrate.
3. the assay method of Rynaxypyr residual quantity in a kind of fruits and vegetables according to claim 1, it is characterized in that, sodium chloride need be added when adopting acetonitrile to extract in step (1) to saltout, sodium acetate need be added when adopting the acetonitrile solution containing 1% acetic acid to extract and saltout.
4. the assay method of Rynaxypyr residual quantity in a kind of fruits and vegetables according to claim 1, it is characterized in that, the agent of step (2) mesostroma dispersive solid-phase extraction is by anhydrous magnesium sulfate, C
18with PSA composition, anhydrous magnesium sulfate, C in every volume extract
18150mg, 50mg and 25mg is respectively with PSA addition.
5. the assay method of Rynaxypyr residual quantity in a kind of fruits and vegetables 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, then rises 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 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: 278,279,280.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106526009A (en) * | 2016-10-24 | 2017-03-22 | 连云港市农村能源环境保护办公室 | Quantitative determination method for general chlorantraniliprole residual quantity |
| CN106546675A (en) * | 2016-10-24 | 2017-03-29 | 苏州市农业科学院 | The quantitative detecting method of Rynaxypyr residual quantity in a kind of tealeaves |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102827145A (en) * | 2012-04-16 | 2012-12-19 | 江苏扬农化工股份有限公司 | Novel deuterated o-aminobenzamide compound, and preparation method and application thereof |
| CN103512993A (en) * | 2013-10-12 | 2014-01-15 | 崔淑华 | Hot pepper and determining method for 96 pesticide residues in product of hot pepper |
-
2014
- 2014-12-30 CN CN201410843724.2A patent/CN104459001B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102827145A (en) * | 2012-04-16 | 2012-12-19 | 江苏扬农化工股份有限公司 | Novel deuterated o-aminobenzamide compound, and preparation method and application thereof |
| CN103512993A (en) * | 2013-10-12 | 2014-01-15 | 崔淑华 | Hot pepper and determining method for 96 pesticide residues in product of hot pepper |
Non-Patent Citations (4)
| Title |
|---|
| SERGIO C. NANITA ET AL: "Fast Extraction and Dilution Flow Injection Mass Spectrometry Method for Quantitative Chemical Residue Screening in Food", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| XIAOSHUI CHEN ET AL: "Development and Validation of a Method for the Determination of 159 Pesticide Residues in Tobacco by Gas Chromatography−Tandem Mass Spectrometry", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| XIAOSHUI CHEN ET AL: "Development and Validation of a Method for the Determination of 159 Pesticide Residues in Tobacco by Gas Chromatography−Tandem Mass Spectrometry", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》, vol. 61, no. 24, 6 June 2013 (2013-06-06) * |
| 朱建华 等: "LC-MS/MS法测定蔬菜和水果中唑虫酰胺、氟啶虫酰胺、氯虫苯甲酰胺和氟虫双酰胺残留量", 《宁波农业科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106526009A (en) * | 2016-10-24 | 2017-03-22 | 连云港市农村能源环境保护办公室 | Quantitative determination method for general chlorantraniliprole residual quantity |
| CN106546675A (en) * | 2016-10-24 | 2017-03-29 | 苏州市农业科学院 | The quantitative detecting method of Rynaxypyr residual quantity in a kind of tealeaves |
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