CN104502506B - A kind of GC-NCI-MS assay method of spiral shell worm ethyl ester residual quantity - Google Patents
A kind of GC-NCI-MS assay method of spiral shell worm ethyl ester residual quantity Download PDFInfo
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Abstract
The invention discloses a kind of GC-NCI-MS assay method of spiral shell worm ethyl ester residual quantity, the method is mainly used in the method measuring spiral shell worm ethyl ester content residual in the complex matrices such as Cereals, animal derived food food agricultural product.Spiral shell worm ethyl ester residual in sample is extracted, C with acetonitrile or containing the acetonitrile solution homogeneous of 1% acetic acid
18after/PSA Solid-Phase Extraction column purification is concentrated, Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detects, and adopts and does not set up the typical curve corrected, quantified by external standard method containing the vehicle solution of agricultural chemicals to be measured.This method average recovery rate is 86.5% ~ 91.7%, average relative standard's deviation (RSD) is 4.1% ~ 7.6%, detection limit lower than 4.72 μ g/kg, have easy and simple to handle, quick, impurity elimination is effective, highly sensitive, characteristic is strong, reproducible, qualitative, quantitative advantage accurately.The U.S., European Union, Japan and other countries being met to the technical requirement of corresponding food safety detection residue limits, providing strong technical support by for ensureing that our people's food security and export abroad trade develop in a healthy way.
Description
Technical field
The present invention relates to a kind of GC-NCI-MS assay method of spiral shell worm ethyl ester residual quantity, be more particularly the method adopting Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) qualitative, quantitative to measure spiral shell worm ethyl ester content residual in the animals and plants derived food of the complex matrices such as animal muscle and goods such as Cereals, pork, beef, mutton, chicken, belong to the determination techniques field of persticide residue.
Background technology
Spiral shell worm ethyl ester (Spirotetramat) is the new tetronic acid derivatives insecticides of Bayer A.G's Development and Production, and insecticidal spectrum is wide, and the lasting period is long.Fatty biosynthesizing mainly through interference insect causes dead larvae, reduces the fertility of adult.Due to the mechanism of action of its uniqueness, effectively can prevent and treat the insect existing pesticide being produced to resistance, can be used as the important kind of nicotinic insecticide resistance management simultaneously.Spiral shell worm ethyl ester is the only modern pesticides had at xylem and the two-way Uptake and translocation performance of bast so far.This compound upwards can move down in whole plant, arrives at blade face and bark, thus prevents the insect on hiding on romaine lettuce and Chinese cabbage internal lobe and fruits and vegetables skin.The interior absorption of this uniqueness can protect newly sprout, leaf and root, prevent ovum and the larval growth of insect.Two-way Uptake and translocation means that insect does not have the place that can hide of safety, and preventive and therapeutic effect is more thorough.Therefore, this agricultural chemicals has good application prospect.Chemical name is cis-4-(ethoxy carbonyl oxygen base)-8-methoxyl-3-(2,5-xylyl)-1-azaspiro [4.5]-3-in last of the ten Heavenly stems alkene-2-ketone.English language Chemical name is called cis-4-(ethoxycarbonyloxy)-8-methoxy-3-(2,5-xylyl)-1-azaspiro [4.5] dec-3-en-2-one.CAS accession number is 382608-10-8, and molecular weight is 373.5, and structural formula is:
Spiral shell worm ethyl ester submits registration to 69 countries and regions.Within 2008, spiral shell worm ethyl ester is successful in the U.S., Canada, Austria, New Zealand, Morocco, Turkey and Tunisia registration.Now obtain registration in more countries and regions successively.Spiral shell worm ethyl ester can be used for various crop, comprises the various suckings pest of control such as cotton, soybean, oranges and tangerines, tropical fruit tree, nut, grape, hops, potato and vegetables, as aphid, thrips, wood louse, aleyrodid, mealybug and scale insect etc.Its to important beneficial insect as ladybug, wasp fly and parasitic wasp have good selectivity.
Along with the registration of spiral shell worm ethyl ester, popularization and use, as U.S. in China's food agricultural product main exit market and European Union, in Cereals and animal derived food, residue limits standard is formulated to it.Environmental Protection Agency issues circular, and announcing draws up has determined in following food/the residual license limitation [calculating by spirotetramat] of table pesticide-spirotetramat: ox, goat, pig, sheep, horse lean meat: 0.01mg/kg; Ox, goat, pig, sheep, horse fat meat: 0.01mg/kg; Ox, goat, pig, sheep, horse liver: 0.01mg/kg; Ox except liver, goat, pig, sheep, horseflesh secondary product: 0.02mg/kg; Regulation has all been made to the maximum maximum permission quantity of spiral shell worm ethyl ester (MRL) in oil crops, cereal, animal & animal product by European Union, and residue limits is from 0.01mg/kg to 0.7mg/kg.
Up to now, have no and the research of spiral shell worm ethyl ester determination of residual amount method in Cereals and animal derived food is reported, liquid chromatography (LC) is all adopted to the detection method of spiral shell worm ethyl ester residual quantity in fruits and vegetables, Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) measures spiral shell worm ethyl ester residual quantity, 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, spiral shell worm ethyl ester belongs to electronegativity compound, but have no the report of the GC-NCI-MS detection method of spiral shell worm ethyl ester residual quantity in vegetables and fruit up to now, due to Cereals, the food agricultural product matrix more complicated such as animal derived food, the good sample-pretreating method of clean-up effect must be set up and instrumental conditions could meet testing requirement, therefore, the detection method setting up spiral shell worm ethyl ester residual quantity in Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) qualitative and quantitative analysis Cereals and animal derived food is significant.
Summary of the invention
The object of this invention is to provide a kind of GC-NCI-MS assay method of spiral shell worm ethyl ester residual quantity, be mainly used in measuring spiral shell worm ethyl ester residual quantity in the complex matrices such as Cereals, animal derived food food agricultural product.
For realizing above object, the technical solution adopted in the present invention is: a kind of GC-NCI-MS assay method of spiral shell worm ethyl ester residual quantity, comprises the steps:
(1) extract
Take mixing sample in tool plug centrifuge tube, after adding suitable quantity of water recovery, quantitatively add acetonitrile or extract containing the acetonitrile solution homogeneous of 1% acetic acid or oscillating ultrasonic, then adding the one in sodium chloride or sodium acetate and anhydrous magnesium sulfate, centrifugal after violent vortex 1min.
(2) purify
Pipette certain volume sample extracting solution, be concentrated into about 1mL, through C
18/ PSA Solid-Phase Extraction column purification, acetonitrile, collects eluent, is concentrated into after doing, and dissolves constant volume, after crossing film, treat that Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detects with acetone/normal hexane mixed solvent that volume ratio is 1/1.
(3) preparation of standard working solution
When same kind matrix blank sample not containing spiral shell worm ethyl ester is processed by above-mentioned steps (1), (2), obtain sample extraction purification residue, add appropriate solvent and mixed standard solution, vortex mixes, and is mixed with the spiral shell worm ethyl ester 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 spiral shell worm ethyl ester in sample liquid, substitute into typical curve, obtain spiral shell worm ethyl ester content in sample liquid, then the Mass Calculation of sample representated by liquid obtains spiral shell worm ethyl ester residual quantity in sample per sample.
Step (1), if the sample of the middle moisture content less such as sample Cereals and animal's liver, must add suitable quantity of water and fully infiltrate before extraction.
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.
Step carries out C in (2)
18/ PSA Solid phase extraction, during acetonitrile, elution volume is 6 ~ 8mL.
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: the ion of Salbutamol Selected Ion Monitoring (SIM) mode monitoring is: 283,89,284.
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 to be applied in Cereals, animal derived food the qualitative confirmation of spiral shell worm ethyl ester in conjunction with GC-NCI-MS and quantitatively to detect, average recovery rate is 86.5% ~ 91.7%, average relative standard's deviation (RSD) is 4.1% ~ 7.6%, detection limit, lower than 4.72 μ g/kg, has easy and simple to handle, quick, accurate, highly sensitive and reproducible advantage.The U.S., European Union, Japan and other countries being met to the technical requirement of corresponding food safety detection residue limits, 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 GC-NCI-MS Selective ion mode chromatogram of Fig. 1 to be concentration be spiral shell worm ethyl ester mark liquid of 100ng/mL.
Fig. 2 is not containing the GC-NCI-MS Selective ion mode chromatogram of the wheat blank sample of spiral shell worm ethyl ester.
Fig. 3 is the GC-NCI-MS Selective ion mode chromatogram of the spiral shell worm ethyl ester be added in blank wheat matrix.
The spiral shell worm ethyl ester standard working curve that Fig. 4 is is substrate preparation with the wheat blank sample not containing spiral shell worm ethyl ester.
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); CR21G III hydro-extractor (Hitachi, Japan); MS3 basic model vortex mixer (IKA, Germany); TurboVapLV type sample automatic concentration instrument (Caliper, USA); 7890N gas chromatography-5975C mass spectrometer (Agilent, USA); C
18/ PSA solid-phase extraction column (6mL, 500mg/500mg) is purchased from Tianjin Bonaaijieer Technology Co., Ltd.
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 spiral shell worm ethyl ester residual quantity in wheat
(1) sample pre-treatments
Extract
Take 5g wheat samples through fully mixing in 50mL centrifuge tube, add the mixing of 5mL water, place 30min, accurately add 20mL acetonitrile, homogeneous extracts 2min, adds 3g anhydrous magnesium sulfate and 2g sodium chloride, after vortex 1min, and the centrifugal 5min of 7000r/min.After centrifugal, get 8mL acetonitrile extract in 40 DEG C revolve steaming or nitrogen blow to about 1mL, to be clean.
Purification
With 5mL acetonitrile prewashing C
18/ PSA solid-phase extraction column, when liquid level arrives the top of adsorbent, said extracted solution is proceeded in post, with 2mL acetonitrile wash test tube, and cleansing solution is moved in SPE post, when solution reaches adsorbent top, add on 4mL acetonitrile to pillar and carry out wash-out, eluent all receives in quantitative test tube, nitrogen dry up rear volume ratio be 1/1 acetone/normal hexane mixed solvent be settled to 1mL, after crossing 0.22 μm of filter membrane, 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 spiral shell worm ethyl ester 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 spiral shell worm ethyl ester in sample liquid, substitute into typical curve, obtain spiral shell worm ethyl ester content in sample liquid, then the Mass Calculation of sample representated by liquid obtains spiral shell worm ethyl ester 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, 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: 283,89,284; Quota ion is 283.
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 spiral shell worm ethyl ester in table 1 wheat bare substrate
Recovery of standard addition and repeatability:
In the wheat not containing spiral shell worm ethyl ester, add the spiral shell worm ethyl ester standard solution of 10, a 20 and 200 μ g/kg3 concentration level, add after 30min until agricultural chemicals and carry out the determination of residual amount by above-mentioned treatment step.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 3 mark-on levels, the average recovery rate of spiral shell worm ethyl ester is 86.9% ~ 90.6%, and average relative standard's deviation (RSD) is 5.1% ~ 7.1%, illustrates that the recovery of the inventive method is higher, reproducible.
The recovery of table 2 spiral shell worm ethyl ester and repeatability (n=6)
Detection limit:
The spiral shell worm ethyl ester matrix standard working solution of variable concentrations is injected GC-NCI-MS, calculate detection limit with the cycles of concentration (cycles of concentration of wheat 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 spiral shell worm ethyl ester is limited to 3.12 μ g/kg.
Embodiment 2: the detection of spiral shell worm ethyl ester residual quantity in pork
(1) sample pre-treatments
Extract
Take 5g pork sample (grinding to form flour) through fully mixing in 50mL centrifuge tube, after adding 20mL water recovery 30min, accurately add the acetonitrile solution of 20mL containing 1% acetic acid, mechanical shaking extraction 20min, ultrasonic extraction 5min, add 3g anhydrous magnesium sulfate and 2g sodium acetate, after vortex 1min, the centrifugal 5min of 7000r/min.After centrifugal, get 8mL acetonitrile extract and revolve steaming or nitrogen blows near dry in 40 DEG C, after adding 1mL acetonitrile vortex, to be clean.
Purification
With 5mL acetonitrile prewashing C
18/ PSA solid-phase extraction column, when liquid level arrives the top of adsorbent, said extracted solution is proceeded in post, with 2mL acetonitrile wash test tube, and cleansing solution is moved in SPE post, when solution reaches adsorbent top, add on 4mL acetonitrile to pillar and carry out wash-out, eluent all receives in quantitative test tube, nitrogen dry up rear volume ratio be 1/1 acetone/normal hexane mixed solvent be settled to 1mL, after crossing 0.22 μm of filter membrane, treat that GC-NCI-MS measures.
(2) preparation of standard working solution
By 100ng/mL standard solution volume ratio be 1/1 acetone/normal hexane mixed solvent be diluted to 10ng/mL standard intermediate liquid, by 10 μ g/mL standard solution dilution be made into 5,2,1,0.5,0.2,0.1 μ g/mL standard solution.Pork 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
Operation steps, chromatogram are consistent with the mensuration of spiral shell worm ethyl ester in above-mentioned pork sample with Mass Spectrometry Conditions.
Qualitative Identification: consistent with the mensuration of spiral shell worm ethyl ester in above-mentioned pork 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=5.603X-68.447, and related coefficient is 0.9997.
Recovery of standard addition and repeatability:
The spiral shell worm ethyl ester standard solution of 10, a 20 and 200 μ g/kg3 concentration level is added in the pork not containing spiral shell worm ethyl ester, add after 30min until agricultural chemicals and carry out the determination of residual amount by above-mentioned treatment step, 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 3 mark-on levels, the average recovery rate of spiral shell worm ethyl ester is 86.5% ~ 91.7%, and average relative standard's deviation (RSD) is 4.1% ~ 7.6%, illustrates that the recovery of the inventive method is high, reproducible.
The recovery of table 3 spiral shell worm ethyl ester and repeatability (n=6)
Detection limit:
The spiral shell worm ethyl ester matrix standard working solution of variable concentrations is injected GC-NCI-MS, calculate detection limit with the cycles of concentration (cycles of concentration of pork 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 spiral shell worm ethyl ester is limited to 4.72 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 (1)
1. a GC-NCI-MS assay method for spiral shell worm ethyl ester residual quantity, is characterized in that, said method comprising the steps of:
(1) sample pretreating: take mixing sample in tool plug centrifuge tube, after adding suitable quantity of water, add acetonitrile or after extracting containing the acetonitrile solution homogeneous of 1% acetic acid or oscillating ultrasonic, add the one in sodium chloride or sodium acetate and anhydrous magnesium sulfate, centrifugal after violent vortex 1min, after centrifugal, pipette certain volume sample extracting solution, be concentrated into about 1mL, through C
18/ PSA Solid-Phase Extraction column purification, acetonitrile, collects eluent, is concentrated into after doing, and dissolves constant volume, after crossing film, treat that Gas Chromatography-Negative chemical ionization source-mass spectrum (GC-NCI-MS) detects with acetone/normal hexane mixed solvent that volume ratio is 1/1;
(2) detect: adopt Gas Chromatography-Negative chemical ionization source-mass spectroscopy, wherein:
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: 283,89,284;
(3) preparation of standard working solution
Processed by above-mentioned steps (1) by same kind matrix blank sample not containing spiral shell worm ethyl ester, obtain sample extraction purification residue, add appropriate solvent and standard solution, vortex mixes, and is mixed with the spiral shell worm ethyl ester series 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 (1) is injected GC-NCI-MS to measure, record the chromatographic peak area of spiral shell worm ethyl ester in sample liquid, substitute into extraction standard working curve, obtain spiral shell worm ethyl ester content in sample liquid, then the Mass Calculation of sample representated by liquid obtains spiral shell worm ethyl ester residual quantity in sample per sample.
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