CN105717222A - GC-EI-MS method for determining residual aminobenzene pyrazolone amount - Google Patents

GC-EI-MS method for determining residual aminobenzene pyrazolone amount Download PDF

Info

Publication number
CN105717222A
CN105717222A CN201610070304.4A CN201610070304A CN105717222A CN 105717222 A CN105717222 A CN 105717222A CN 201610070304 A CN201610070304 A CN 201610070304A CN 105717222 A CN105717222 A CN 105717222A
Authority
CN
China
Prior art keywords
amine
phenopyrazone
sample
acetonitrile
solution
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201610070304.4A
Other languages
Chinese (zh)
Inventor
郭庆龙
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to CN201610070304.4A priority Critical patent/CN105717222A/en
Publication of CN105717222A publication Critical patent/CN105717222A/en
Pending legal-status Critical Current

Links

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

Landscapes

  • 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 GC-EI-MS method for determining the residual aminobenzene pyrazolone amount.The method is mainly used for determining the residual aminobenzene pyrazolone amount in cereal grains, animal derived foods and other complicated matrix foods or agricultural products.Acetonitrile or an acetonitrile solution containing 1% acetic acid are adopted to homogeneously extract the residual aminobenzene pyrazolone in a sample, purification by adopting a C18/PSA solid-phase extraction column and concentration are performed, then gas chromatography-electron impact ionization-mass spectrometry (GC-EI-MS) detection is performed, a blank matrix solution containing no pesticide to be detected is adopted to establish a standard working curve for correction, and quantification is performed by adopting an external standard method.The average recovery rate of the method is 90.5%-94.7%, the average relative standard deviation (RSD) of the method is 5.9%-8.9%, the detection limit of the method reaches 2.14 micrograms/kilogram or below, and the method has the advantages of being simple and convenient to operate, rapid, good in impurity removal effect, high in sensitivity, good in characteristic performance and repeatability and accurate in qualititation and quantification, can meet the technical requirements for corresponding product safety detection of the United States, European Union, Japan and other countries and provides a powerful technical support for ensuring the people's food safety and healthy development of the abroad export trade of our country.

Description

A kind of GC-EI-MS assay method of amine Phenopyrazone residual quantity
Technical field
The present invention relates to the GC-EI-MS assay method of a kind of amine Phenopyrazone residual quantity, more specifically use gas chromatogram -electron impact ion source-mass spectrum (GC-EI-MS) qualitative, quantitative measures the animal fleshes such as Cereals, Carnis Sus domestica, beef, Carnis caprae seu ovis, Carnis Gallus domesticus In the animals and plants derived food of the complex matrices such as meat and goods, the method for the amine Phenopyrazone content of residual, belongs to persticide residue Determination techniques field.
Background technology
Amine Phenopyrazone (Fenpyrazamine) is the heterocyclic based on pyridine of SUMITOMO CHEMICAL KCC research and development Antifungal, by the pathogen of the mechanism of action of suppression ergosterol biosynthesis pathway, demonstrates mycelial growth and spore Sprout and the inhibitory action of pollen tube growth.No. CAS is: 473798-59-3, and molecular formula is C17H21N3O2S, molecular weight is 331.43, the chemical structural formula of amine Phenopyrazone is as follows:
Amine benzene pyrrole bacterium ketone is the novel germicide of Sumitomo Chemical stand-alone development, has novel chemical constitution, can rapidly permeate into and make object In, to the fungus energy fast onset drug effect infected, it is mainly used in gray mold, the sclerotium controlling in Fructus Vitis viniferae, carpolities, vegetable crop Disease and chain sclerotinia sclerotiorum.It addition, amine benzene pyrrole bacterium ketone is to mammal high safety, can degrade rapidly in the environment, can be multiple Crop is applied before gathering.Amine benzene pyrrole bacterium ketone is initially released in Korea S, after the amine benzene pyrrole former medicine of bacterium ketone obtains first whole world registration, Sumitomo Chemical releases its antibacterial new product containing the amine benzene pyrrole former medicine of bacterium ketone in Korea S again, and sells with Botrycide trade (brand) name, This product is the suspending agent type of 30%, for fruits and vegetables crop.2009, Sumitomo Chemical was used for temperature in the registration of application European Union The vegetable crops such as Fructus Lycopersici esculenti, Fructus Solani melongenae, Fructus Capsici and the calabash of room cultivation, the also Field information of Fructus Vitis viniferae, but the most only obtain in Poland Must register for 3 years by a definite date, trade name Prolectu temporarily.In June, 2012, European Union's food and animal health standing committee Having voted its registration application through, amine benzene pyrrole bacterium ketone has obtained European Union's approval registration, will be formal in 1 day January in 2013 Listing the registered effective ingredient list under European Union's pesticide 1107/2009 regulation in, effect duration is 10 years.Amine benzene pyrrole bacterium ketone is in European Union After obtaining registration, Fructus Vitis viniferae and some chamber crops will be used for, such as Fructus Lycopersici esculenti, Fructus Solani melongenae, Fructus Capsici and calabash etc..Amine benzene pyrrole bacterium ketone is to Portugal Grape spore bacterium has an excellent prevention and control ability, Sumitomo Chemical will further in 2012/13 year to Italy and Chile's this product of marketing Product, its whole world trade name Prolectus.Registered in the U.S. and a lot of Asian countries, be there is the most wide application Prospect.
Along with amine Phenopyrazone registration, promote and use, as European Union in China's main exit market, the U.S., Canada Its maximum allowable residual quantity (MRL) in the food agricultural product such as veterinary antibiotics, Cereals and livestock products has been formulated in country, European Union formulates antibacterial amine benzene pyrrole bacterium ketone MRL standard in various crop.This standard is on May 7th, 2012 Come into force.Amine benzene pyrrole bacterium ketone MRL in Rhizoma Solani tuber osi, Fructus Capsici, Fructus Solani melongenae is 3mg/kg；At calabash, Fructus Cucumidis sativi, little Huang MRL in melon, Courgettes is 0.7mg/kg；In the fruit such as Citrus, grapefruit, Fructus Citri Limoniae and various dry fruit Residue limits is most stringent, for 0.01mg/kg.The U.S. specifies that amine Phenopyrazone MRL in Semen Armeniacae Amarum is 0.02 mg/kg；MRL in Semen Armeniacae Amarum shell is 1.5mg/kg；MRL in Fructus Pistaciae Verae is 0.02mg/kg； MRL in lettuce heads is 1.5mg/kg；MRL in leaf lettuce is 2mg/kg；Sub-cranberry MRL in group 13-07A is 5mg/kg；MRL in bush fruit class subgroup 13-07B is 5 mg/kg；In small-sized fruit 13-07F subgroup of overgrowing, the MRL of (in addition to Fructus actinidiae chinensis) is 3mg/kg；At Fructus Vitis viniferae MRL in juice is 4mg/kg；MRL in short youngberry class subgroup 13-07G is 3mg/kg；? MRL in Radix Ginseng is 0.7mg/kg.Canada regulation amine Phenopyrazone Sucus Vitis viniferae, Caulis et Folium Lactucae Sativae, Caulis et Folium Lactucae sativae, Radix Ginseng, M RL in Semen Armeniacae Amarum nut, Fructus Pistaciae Verae is respectively 4.0mg/kg, 2.0mg/kg, 1.5mg/kg, 0.7mg/kg and 0.02mg/kg； When European Union, Japan and other countries specify the residue limits standard not formulating corresponding pesticide, it is exported to the food agricultural product bag of its country Include residue limits in the animal derived foods such as livestock meat and all carry out " uniform limit " of 0.01mg/L.
Present stage, the research to amine Phenopyrazone determination of residual amount method is less, and the detection method of report is mainly vegetable and water Amine Phenopyrazone method for detecting residue in Guo, these detection methods all use Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) to measure The detection method of amine Phenopyrazone residual quantity in vegetable and fruit, uses LC-MS/MS to measure food Residual Pesticides in Farm Produce tool There is quick, easy, sensitivity advantages of higher, but due to its price costly, a lot of testing agencies, enterprise or scientific research institutions are not Configure this instrument or configuration number of units is less, when using LC-MS/MS detection due to different compounds, different flowings need to be used Mutually or chromatographic column, such needs are constantly changed chromatographic column, flowing and mutually and are expended the long time and be balanced system, this Determine to constrain in degree the application of LC-MS/MS.The gaschromatographic mass spectrometry (GC-EI-MS) being equipped with electron impact ionization source is analyzed Food Residual Pesticides in Farm Produce tool has great advantage, and owing to electron impact ionization source mass spectrum is versatility detector, can realize several The multi-residue analysis of hundred kinds of pesticide, can be the most qualitative and quantitative, moderate, and therefore existing various testing agencies and enterprise are equipped with Pesticide residues in food agricultural product are detected by gas chromatogram-electron impact ion source-mass spectrograph (GC-EI-MS), but so far Having no the report of the GC-EI-MS detection method of amine Phenopyrazone residual quantity in food agricultural product till the present, it is negative that amine Phenopyrazone belongs to electricity Property compound, owing to the food agricultural product substrate such as Cereals, animal derived food is more complicated, must set up the sample that clean-up effect is good Product pre-treating method and instrumental conditions could meet testing requirement, therefore, set up gas chromatogram-electron impact ion source-matter In spectrum (GC-EI-MS) qualitative and quantitative analysis Cereals and animal derived food, the detection method of amine Phenopyrazone residual quantity has weight Want meaning.
Summary of the invention
It is an object of the invention to provide the GC-EI-MS assay method of a kind of amine Phenopyrazone residual quantity, be mainly used in measuring grain Amine Phenopyrazone residual quantity in the complex matrices food agricultural product such as paddy, animal derived food.
For realizing object above, the technical solution adopted in the present invention is: the GC-EI-MS of a kind of amine Phenopyrazone residual quantity Assay method, comprises the steps:
(1) extract
Weigh mixing sample in tool plug centrifuge tube, after adding suitable quantity of water recovery, be quantitatively adding acetonitrile or equal containing the acetonitrile solution of 1% acetic acid Matter or oscillating ultrasonic extract, and are subsequently 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 C18/ PSA Solid-Phase Extraction column purification, acetonitrile eluting, collection is washed De-liquid, after being concentrated to dryness, dissolves constant volume with the acetone that volume ratio is 1/1/normal hexane mixed solvent, after crossing film, treat gas chromatogram- Electron impact ion source-mass spectrum (GC-EI-MS) detects.
(3) preparation of standard working solution
When same kind matrix blank sample without amine Phenopyrazone is processed by above-mentioned steps (1), (2), obtain sample extraction and purify Residue, adds appropriate solvent and standard solution, and vortex mixes, and is configured to the amine Phenopyrazone series standard work of at least 3 concentration Liquid.
(4) gas chromatogram-electron impact ion source-mass spectrography (GC-EI-MS) measures
The standard working solution of each Concentraton gradient in step (3) is carried out GC-EI-MS mensuration, with the chromatographic peak area of standard working solution Its respective concentration is carried out regression analysis, obtains standard working curve；Sample after purifying in step (2) under the same conditions Liquid injects GC-EI-MS and is measured, and records the chromatographic peak area of amine Phenopyrazone in sample liquid, substitutes into standard working curve, Amine Phenopyrazone content in sample liquid, then obtains amine Phenopyrazone in sample according to the Mass Calculation of sample representated by sample liquid Residual quantity.If amine Phenopyrazone residual quantity exceedes the range of linearity upper limit in upper machine solution, need to be with constant volume solvent by upper machine solution concentration Within being diluted to the range of linearity.
Step (1) if in the sample of the moisture content less such as sample Cereals and animal livers, suitable quantity of water must be added before extraction abundant Infiltration.
Add sodium chloride when step (1) uses acetonitrile extraction to saltout, use the acetonitrile solution containing 1% acetic acid to add when extracting Sodium acetate is saltoutd.
Step carries out C in (2)18/ PSA Solid phase extraction, during acetonitrile eluting, elution volume is 6～8mL.
In step (4), GC conditions is: chromatographic column: HP-5MS capillary chromatographic column, column length 30m, internal diameter 0.25 Mm, thickness 0.25 μm；Injector temperature 250 DEG C；Carrier gas: He, not shunt mode sample introduction, sample size: 1 μ L；Constant current mould Formula, 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, so After rise to 220 DEG C with the speed of 2 DEG C per minute, then rise to 280 DEG C with the speed of 20 DEG C per minute, keep 10min；Transmission line temperature Degree: 280 DEG C.
In step (4), Mass Spectrometry Conditions is: ion source temperature 150 DEG C；Quadrupole rod temperature 150 DEG C；Ionization pattern: electronics bangs Hit ionization, i.e. EI pattern, energy 70eV；Scan mode: Salbutamol Selected Ion Monitoring (SIM) pattern, the ion of monitoring is: 230.1, 331.1、256.1、231.1。
When step (4) measures sample liquid and extraction standard working solution, if sample liquid Pesticides chromatographic peak retention time and standard In solution, corresponding pesticide retention time is consistent, and in the sample mass spectrum after background correction, selected ion all occurs, And abundance of ions is more consistent than the abundance of ions ratio with standard solution, then can determine whether sample liquid exists this pesticide；If above-mentioned two Individual condition can not meet simultaneously, then judge without this kind of pesticide.
The beneficial effects of the present invention is:
The present invention utilizes dispersive solid-phase extraction technology, establishes simplicity, quickly and can be prevented effectively from the sample of sample mesostroma interference Product pre-treating method, this pre-treating method combines GC-EI-MS, and to be applied to amine Phenopyrazone in Cereals, animal derived food qualitative Confirmation and detection by quantitative, average recovery rate is 90.5%～94.7%, and average relative standard's deviation (RSD) is 5.9%～8.9%, Detection limit is less than 2.14 μ g/kg, has easy and simple to handle, quick, accurate, highly sensitive and reproducible advantage.U.S. can be met State, European Union, the Japan and other countries technology requirement to corresponding product safety detection, for ensureing our people's food safety and to going out Mouth trade sound development provides strong technical support.
Accompanying drawing explanation
Fig. 1 is that the GC-EI-MS being added on the amine Phenopyrazone in blank Carnis Sus domestica substrate selects chromatography of ions figure.
Fig. 2 is that the GC-EI-MS of the Carnis Sus domestica blank sample without amine Phenopyrazone selects chromatography of ions figure.
Fig. 3 is the amine Phenopyrazone standard working curve with the Carnis Sus domestica blank sample without amine Phenopyrazone as substrate preparation.
Detailed description of the invention
Now with following embodiment, the present invention is described, but is not to limit the scope of the present invention.
The instrument used in embodiment and reagent
T18Basic homogenizer (IKA, Germany)；CR21G III centrifuge (Hitachi, Japan)；MS3 basic model is revolved Whirlpool blender (IKA, Germany)；TurboVap LV pattern product automatic concentration instrument (Caliper, USA)；7890N gas phase color Spectrum-5977C mass spectrograph (Agilent, USA)；C18/ PSA solid-phase extraction column (6mL, 500mg/500mg) is purchased from Tianjin and wins Na Aijieer Science and Technology 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 analytical pure, are purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Standard substance: purity 98.0%, purchased from Japan and Guang Chun agro-industry Co., Ltd..
Embodiment 1: the detection of amine Phenopyrazone residual quantity in Semen Tritici aestivi
(1) sample pre-treatments
Extract
Weigh through the abundant 5g wheat samples (grinding to form flour) mixed in 50mL centrifuge tube, after adding 20mL water recovery 30min, Accurately add the 20mL acetonitrile solution containing 1% acetic acid, mechanical shaking extraction 20min, supersound extraction 5min, add 3g anhydrous magnesium sulfate With 2g sodium acetate, after vortex 1min, 7000r/min is centrifuged 5min.After Li Xin, take 8mL acetonitrile extraction liquid in 40 DEG C rotation steam or Nitrogen is blown near dry, after adding 1mL acetonitrile vortex, to be clean.
Purify
With 5mL acetonitrile prewashing C18/ PSA solid-phase extraction column, when liquid level arrives the top of adsorbent, proceeds to post by said extracted solution In, wash test tube with 2mL acetonitrile, and cleaning mixture is moved in SPE post, when solution reaches adsorbent top, add 4mL Acetonitrile carries out eluting to pillar, and eluent all receives in quantitative test tube, nitrogen dry up after with the acetone that volume ratio is 1/1/ Normal hexane mixed solvent is settled to 1mL, after crossing 0.22 μm filter membrane, treats that GC-EI-MS measures.
(2) preparation of standard working solution
Accurately weighing 25 ± 0.1mg standard substance in 25mL volumetric flask, dissolve with acetonitrile, constant volume obtains 1000.0 μ g/mL standard reserving solutions； Pipette 1.0mL standard reserving solution to be placed in 100mL volumetric flask, obtain with the acetone that volume ratio is 1/1/normal hexane mixed solvent constant volume To 10.0 μ g/mL standard intermediate liquids；10 μ g/mL standard solution dilutions are made into 5,2,1,0.5,0.2,0.1 μ g/mL mark Quasi-solution.Semen Tritici aestivi blank sample without amine Phenopyrazone is processed by above-mentioned pre-treatment step, obtains sample extraction and purify residue, Adding acetone/normal hexane mixed solvent and the 100 above-mentioned standard solution of μ L that 900 μ L volume ratios are 1/1 in this residue, vortex mixes Even, it is made into 10,20,50,100,200,500 μ g/L extraction standard working solutions.
(3) gas chromatogram-electron impact ion source-mass spectrography (GC-EI-MS) measures
The standard working solution of variable concentrations gradient is injected separately into GC-EI-MS, carries out quantitatively dividing of amine Phenopyrazone content with external standard method Analysis, i.e. carries out regression analysis with the chromatographic peak area of standard working solution to its respective concentration, obtains standard working curve；Identical Under the conditions of sample extracting solution injected GC-EI-MS be measured, record the chromatographic peak area of amine Phenopyrazone in sample liquid, substitute into Standard working curve, obtains amine Phenopyrazone content in sample liquid, then obtains according to the Mass Calculation of sample representated by sample liquid Amine Phenopyrazone residual quantity in 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.
Furnace box heating schedule: initial temperature 60 DEG C keeps 2min, rises to 200 DEG C with the speed of 20 DEG C per minute, then with per minute The speed of 2 DEG C rises to 220 DEG C, then rises to 280 DEG C with the speed of 20 DEG C per minute, keeps 10min.
Transmission line temperature: 280 DEG C.
Wherein, mass spectrometry parameters is:
Ionization pattern: electron impact ionization, i.e. EI 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: 230.1,331.1,256.1,231.1； Quota ion is 230.1.
Qualitative Identification: at identical conditions, if sample liquid Pesticides chromatographic peak retention time pesticide corresponding to standard solution Retention time is consistent, and in the sample mass spectrum after background correction, selected ion all occurs, and abundance of ions Ratio is consistent with the abundance of ions ratio of standard solution, then can determine whether to exist in sample liquid this pesticide；If above-mentioned two condition can not be same Time meet, then judge without this kind of pesticide.
With the chromatographic peak area of standard working solution, its respective concentration is carried out regression analysis, obtain standard working curve such as table 1.
The standard working curve of amine Phenopyrazone in table 1 Semen Tritici aestivi bare substrate
Title Retention time (min) Regression equation Correlation coefficient
Amine Phenopyrazone 26.27 Y=879.32X-606.04 0.9998
Recovery of standard addition and repeatability:
The amine Phenopyrazone standard of 10,20 and 200 g/kg3 concentration levels of μ is added in the Semen Tritici aestivi without amine Phenopyrazone Solution, carries out the determination of residual amount by above-mentioned process step after pesticide adds 30min.Mensuration concentration is added dense with pesticide theory Degree compares, and obtains pesticide TIANZHU XINGNAO Capsul, each pitch-based sphere parallel assay 6 times, obtains its relative standard deviation, measures The results are shown in Table 2.As can be seen from Table 2, in 3 mark-on levels, the average recovery rate of amine Phenopyrazone is 90.5%～94.7%, Average relative standard's deviation (RSD) is 5.9%～8.0%, illustrates that the response rate of the inventive method is higher, reproducible.
The response rate of table 2 amine Phenopyrazone and repeatability (n=6)
Detection limit:
The amine Phenopyrazone extraction standard working solution of variable concentrations is injected GC-EI-MS, molten with least concentration extraction standard 3 times of signal to noise ratios at liquid chromatography peak and the cycles of concentration (cycles of concentration of Semen Tritici aestivi is 2.0 times) of sample handling processes calculate detection limit, The detection of amine Phenopyrazone is limited to 1.69 μ g/kg.
Embodiment 2: the detection of amine Phenopyrazone residual quantity in Carnis Sus domestica
(1) sample pre-treatments
Extract
Weigh through the abundant 5g Carnis Sus domestica sample mixed in 50mL centrifuge tube, add the mixing of 5mL water, place 30min, accurately add 20mL acetonitrile, homogenizing extracts 2min, adds 3g anhydrous magnesium sulfate and 2g sodium chloride, and after vortex 1min, 7000r/min is centrifuged 5min.After Li Xin, take 8mL acetonitrile extraction liquid and be blown to about 1mL in 40 DEG C of rotation steamings or nitrogen, to be clean.
Purify
With 5mL acetonitrile prewashing C18/ PSA solid-phase extraction column, when liquid level arrives the top of adsorbent, proceeds to post by said extracted solution In, wash test tube with 2mL acetonitrile, and cleaning mixture is moved in SPE post, when solution reaches adsorbent top, add 4mL Acetonitrile carries out eluting to pillar, and eluent all receives in quantitative test tube, nitrogen dry up after with the acetone that volume ratio is 1/1/ Normal hexane mixed solvent is settled to 1mL, after crossing 0.22 μm filter membrane, treats that GC-EI-MS measures.
(2) preparation of standard working solution
Acetone/normal hexane mixed solvent that 1000 μ g/mL standard solution volume ratios are 1/1 is diluted to 10 μ g/mL standard intermediate liquids, will 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 pig without amine Phenopyrazone Meat blank sample is processed by above-mentioned pre-treatment step, obtains sample extraction and purifies residue, and adding 900 μ L volume ratios in this residue is The acetone of 1/1/normal hexane mixed solvent and the 100 above-mentioned standard solution of μ L, vortex mixes, be made into 10,20,50,100,200, 500 μ g/L extraction standard working solutions.
(3) gas chromatogram-electron impact ion source-mass spectrography (GC-EI-MS) measures
Operating procedure, chromatograph are consistent with the mensuration of amine Phenopyrazone in above-mentioned wheat samples with Mass Spectrometry Conditions.
Qualitative Identification: consistent with the mensuration of amine Phenopyrazone in above-mentioned wheat samples.
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=1117.3X-4751.1, correlation coefficient is 0.9994.
Recovery of standard addition and repeatability:
The amine Phenopyrazone standard of 10,20 and 200 3 concentration levels of μ g/kg is added in the Carnis Sus domestica without amine Phenopyrazone Solution, carries out the determination of residual amount by above-mentioned process step after pesticide adds 30min, adds dense by mensuration concentration with pesticide theory Degree compares, and obtains pesticide TIANZHU XINGNAO Capsul, each pitch-based sphere parallel assay 6 times, obtains its relative standard deviation, measures The results are shown in Table 3.As can be seen from Table 3, in 3 mark-on levels, the average recovery rate of amine Phenopyrazone is 90.5%～92.3%, Average relative standard's deviation (RSD) is 7.6%～8.9%, illustrates that the response rate of the inventive method is high, reproducible.
The response rate of table 3 amine Phenopyrazone and repeatability (n=6)
Detection limit:
The amine Phenopyrazone extraction standard working solution of variable concentrations is injected GC-EI-MS, molten with least concentration extraction standard 3 times of signal to noise ratios at liquid chromatography peak and the cycles of concentration (cycles of concentration of Carnis Sus domestica is 2.0 times) of sample handling processes calculate detection limit, The detection of amine Phenopyrazone is limited to 1.69 μ g/kg.
Above embodiment is only to be described the preferred embodiment of the present invention, not limits the scope of the present invention Fixed, on the premise of designing spirit without departing from the present invention, it is each that technical scheme is made by this area ordinary skill technology Plant modification and improvement, all should fall in the protection domain that claims of the present invention determines.

Claims (5)

1. the GC-EI-MS assay method of an amine Phenopyrazone residual quantity, 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, add acetonitrile or containing the acetonitrile solution homogenizing of 1% acetic acid or shake After swinging supersound extraction, add 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 C18/ PSA Solid-Phase Extraction column purification, acetonitrile eluting, receive Collection eluent, after being concentrated to dryness, dissolves constant volume with the acetone that volume ratio is 1/1/normal hexane mixed solvent, after crossing film, treats gas phase color Spectrum-electron impact ion source-mass spectrum (GC-EI-MS) detects；
(3) preparation of standard working solution
Same kind matrix blank sample without amine Phenopyrazone is processed by above-mentioned steps (1), (2), obtains sample extraction clean Changing residue, add appropriate solvent and standard solution, vortex mixes, and is configured to the amine Phenopyrazone series standard work of at least 3 concentration Make liquid；
(4) measure and result calculates
The standard working solution of each Concentraton gradient in step (3) is carried out GC-EI-MS mensuration, with the chromatographic peak of standard working solution Area carries out regression analysis to its respective concentration, obtains extraction standard working curve；Step (2) will be purified under the same conditions After sample liquid inject GC-EI-MS and be measured, record the chromatographic peak area of amine Phenopyrazone in sample liquid, substitute into extraction standard Working curve, obtains amine Phenopyrazone content in sample liquid, then obtains sample according to the Mass Calculation of sample representated by sample liquid Middle amine Phenopyrazone residual quantity；If amine Phenopyrazone residual quantity exceedes the range of linearity upper limit in upper machine solution, need to incite somebody to action with constant volume solvent Within upper machine solution concentration is diluted to the range of linearity.
The GC-EI-MS assay method of a kind of amine Phenopyrazone residual quantity the most according to claim 1, it is characterised in that step Suddenly (1) if in the sample of sample moisture content less, suitable quantity of water must be added before extraction and fully infiltrate.
The GC-EI-MS assay method of a kind of amine Phenopyrazone residual quantity the most according to claim 1, it is characterised in that step Sodium chloride need to be added when (1) uses acetonitrile extraction suddenly to saltout, use the acetonitrile solution containing 1% acetic acid need to add acetic acid when extracting Sodium salt is analysed.
The GC-EI-MS assay method of a kind of amine Phenopyrazone residual quantity the most according to claim 1, it is characterised in that step Suddenly (2) carry out C18/ PSA Solid phase extraction, during acetonitrile eluting, elution volume is 6～8mL.
The GC-EI-MS assay method of a kind of amine Phenopyrazone residual quantity the most according to claim 1, it is characterised in that step Suddenly in (4), GC-EI-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 keep 2min, rise to 200 DEG C with the speed of 20 DEG C per minute, then with The speed of 2 DEG C per minute rises to 220 DEG C, then rises to 280 DEG C with the speed of 20 DEG C per minute, keeps 10min；Transmission line temperature: 280℃；Ionization pattern: electron impact ionization, i.e. EI pattern, energy 70eV；Ion source temperature 150 DEG C；Scan mode: choosing Selecting ion monitoring (SIM) pattern, the ion of monitoring is: 230.1,331.1,256.1,231.1.
CN201610070304.4A 2016-01-30 2016-01-30 GC-EI-MS method for determining residual aminobenzene pyrazolone amount Pending CN105717222A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610070304.4A CN105717222A (en) 2016-01-30 2016-01-30 GC-EI-MS method for determining residual aminobenzene pyrazolone amount

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610070304.4A CN105717222A (en) 2016-01-30 2016-01-30 GC-EI-MS method for determining residual aminobenzene pyrazolone amount

Publications (1)

Publication Number Publication Date
CN105717222A true CN105717222A (en) 2016-06-29

Family

ID=56154581

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610070304.4A Pending CN105717222A (en) 2016-01-30 2016-01-30 GC-EI-MS method for determining residual aminobenzene pyrazolone amount

Country Status (1)

Country Link
CN (1) CN105717222A (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014101319A (en) * 2012-11-21 2014-06-05 Nissan Chem Ind Ltd Germicide, nematicide or bactericide compositions
CN104535686A (en) * 2014-12-30 2015-04-22 山东出入境检验检疫局检验检疫技术中心 GC-NCI-MS determination method for cyenopyrafen residual quantity

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014101319A (en) * 2012-11-21 2014-06-05 Nissan Chem Ind Ltd Germicide, nematicide or bactericide compositions
CN104535686A (en) * 2014-12-30 2015-04-22 山东出入境检验检疫局检验检疫技术中心 GC-NCI-MS determination method for cyenopyrafen residual quantity

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
PARK,HYEJI 等: "Development and validation of an analytical method for fungicide fenpyrazamine determination in agricultural products by HPLC-UVD", 《ANALYTICAL SCIENCE & TECHNOLOGY》 *
SUSAN STRANGE HERRMANN 等: "《www.eurl-pesticides.eu/docs/public/tmplt_article.asp?LabID=400&CntID=618&Theme_ID=1&Pdf=False&Lang=EN》", 31 December 2015 *
杨淑娴 等: "胺苯吡菌酮原药的液相色谱分析方法研究", 《农药科学与管理》 *

Similar Documents

Publication Publication Date Title
CN104502515B (en) A kind of LC-MS/MS assay method of four chlorantraniliprole residual quantities
CN104502507A (en) GC-NCI-MS determination method for determining residual amount of chlorantraniliprole
CN104678043B (en) A kind of GC-EI-MS assay method of fluorine ether bacterium amide residual quantity
CN104502484B (en) A kind of GC-NCI-MS measures the method that in fruits and vegetables, cyanogen insect amide is residual
CN104730191B (en) A kind of LC-MS/MS assay method of fluorine ether bacterium amide residual quantity
CN106546675B (en) The quantitative detecting method of Rynaxypyr residual quantity in a kind of tealeaves
CN104458968B (en) The GC-EI-MS assay method of spiral shell worm ethyl ester residual quantity in a kind of fruits and vegetables
CN104502508B (en) A kind of GC-NCI-MS assay method of cyanogen insect amide residual quantity
CN104459000A (en) GC-EI-MS (gas chromatography-electron ionization-mass spectrum) measuring method for chlorantraniliprole residual quantity
CN104655781B (en) A kind of assay method of metrafenone residual quantity
CN105717224A (en) GC-EI-MS method for rapidly determining residual aminobenzene pyrazolone amount
CN104535686B (en) A kind of GC-NCI-MS assay method of nitrile pyrrole mite ester residual quantity
CN104502505B (en) A kind of GC-EI-MS assay method of spiral shell worm ethyl ester residual quantity
CN105717222A (en) GC-EI-MS method for determining residual aminobenzene pyrazolone amount
CN105717211A (en) GC-MS/MS measurement method for penflufen residual amount
CN105548444A (en) GC-MS/MS (gas chromatography-tandem mass spectrometry) determining method of aniline pyrazolone residual quantity
CN105717213A (en) GC-NIC-MS measurement method for penflufen residual amount
CN104569198B (en) A kind of GC-EI-MS assay method of nitrile pyrrole mite ester residual quantity
CN105717219A (en) Method for rapidly determining residual fenpyrazamine in animal-derived foods
CN105717218A (en) GC-MS/MS determining method for residual silthiopham quantity
CN105717221A (en) GC-EI-MS rapid measurement method for penflufen residual amount
CN105548447A (en) GC (Gas Chromatograph)-EI (Electronic Impact Ionization)-MS (Mass Spectrometry) determining method of residual quantity of fluxapyroxad
CN105572288A (en) Method for determining aminobenzene pyrazolone residues in fruits and vegetables through GC-EI-MS
CN105699523A (en) Measurement method for aniline pyrazolone residue in vegetables and fruits
CN105606755A (en) GC-NCI-MS (gas chromatography-negative chemical ionization-mass spectrometry) measuring method for aminobenzene pyrazolone residue

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20160629