CN106770704B - The detection method of cyflufenamid residual quantity in a kind of wheat - Google Patents
The detection method of cyflufenamid residual quantity in a kind of wheat Download PDFInfo
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- CN106770704B CN106770704B CN201611018989.4A CN201611018989A CN106770704B CN 106770704 B CN106770704 B CN 106770704B CN 201611018989 A CN201611018989 A CN 201611018989A CN 106770704 B CN106770704 B CN 106770704B
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- 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
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- 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
Abstract
The invention discloses a kind of detection method of cyflufenamid residual quantity in wheat, step is divided into: 1, the preparation of standard solution；2, standard curve is drawn；3, sample extraction；4, sample purification；5, sample detection；6, result calculates；Detection method of the invention is using acetonitrile as extracting solution, and dosage is less compared with reported acetone is as extracting solution, and later purification is simpler；Compared with methanol is as extracting solution, after extracting, saltouing, heating plate is evaporated, and does not need to rotate, and can improve working efficiency；Detection instrument popularity rate used is higher, and method easily promotes and applies；The reproducibility of detection method of the invention, accuracy, precision and detection limit can meet retention analysis requirement of the pesticide in wheat；Detection method of the invention reduces environmental pollution, reduces testing cost, improves work efficiency, and has stronger practicability, has good application value.
Description
Technical field
The invention belongs to Detecting Pesticide fields, and in particular to the detection side of cyflufenamid residual quantity in a kind of wheat
Method.
Background technique
Cyflufenamid English name: cyflufenamid, No. CAS: 180409-60-3, molecular formula: C20H17F5N2O2,
Chemical name: (Z)-N- [the fluoro- 6-(trifluoromethyl of α-(cyclopropyl methoxyimino) -2,3- bis-) benzyl] -2- phenyl acetamide.Ring
Fluorine bacterium amine category acid amide fungicides, by inhibiting haustorium mitogenetic on mycelia in the Powdery Mildew history of life (namely pathogenic process)
It is formed and is grown, the growth of secondary hyphae and the formation of hapteron, but the court of a feudal ruler of spore germination, germ tube length and hapteron are formed
Without effect.With the fungicide no interactions resistance in current commercialization and positive research, to include to demethylation inhibitor, sterols and
All strains of the resistant Powdery Mildew of benzimidazole germicide are effective.It has protection and therapeutic effect, have evaporation and
Transferance, control efficiency is fine at very low concentrations, it to sterol, phosphonate ester, chitin, Protein synthesis without
Inhibiting effect.The medicine has been widely applied in China as a kind of new type bactericide.The detection method registered at present only has poplar
Cloud tints bamboo fish trap (2008) Gas Chromatography-Negative chemical ionization source mass spectrography detects remaining cyflufenamid and Li Li (2008 in carrot
Year) research of the gas chromatography-mass spectrography detection method of cyflufenamid residual quantity, inspection of the cyflufenamid in wheat in food
Survey method has not been reported.
Extracting solution is mostly acetone in extracting method in the prior art, and dosage is more, and later purification is complicated, and environmental pollution is tight
Weight；After extracting, saltouing, mostly rotary evaporation, working efficiency are low；Some method pre-treatments are simple, but detecting instrument is matter
Spectrum, popularity rate is lower, and it is difficult that method promotes and applies, and testing cost is high.
Summary of the invention
Aiming at the problems existing in the prior art, the present invention provides the remaining detection method of cyflufenamid in a kind of wheat,
Extracting solution dosage is less, and later purification is simpler, and less environmental pollution reduces testing cost；After extracting, saltouing, heating
Plate is evaporated, and is not needed to rotate, be improved work efficiency；Detection instrument used is liquid phase, and popularity rate is higher.Have stronger practical
Property, there is good application value.
In order to solve the above technical problems, the invention adopts the following technical scheme:
The detection method of cyflufenamid residual quantity in a kind of wheat, steps are as follows:
(1) preparation of standard solution: ring not bacterium amine standard items are weighed, are made 1000 mg/L's with n-hexane dissolution constant volume
Mother liquor, then it is diluted to the standard solution of 0.05mg/L, 0.1mg/L, 0.5mg/L, 1mg/L, 2mg/L step by step；
(2) draw standard curve: by standard solution made from step (1) cross chromatographic column RTX-5(30 m × 0.25 mm ×
0.25 μm) it is detected afterwards with ECD detector, injector temperature is 240 DEG C, and detector temperature is 300 DEG C, gas (N2) flow be 2
ML/min, make-up gas (N2) flow is 30mL/min, split ratio 30:1, sample volume is 1 μ L, is 180 DEG C of condition in column temperature
Then lower reservation 2min rises to 260 DEG C of 10 min of reservation according to the heating rate of 20 DEG C/min；
Using the concentration of standard solution as abscissa x, peak area is ordinate y, computer drawing standard curve, standard curve
Linear equation is y=175183.02x+17363.93, wherein x is the concentration of ring not bacterium amine standard solution, unit mg/L, y
For peak area, response related coefficient is r2=0.99；
(3) sample extraction: sample will be made after wheat crushing, weighs 10.00 g of the sample in 100 mL centrifugal bottles, adds
10 mL water and 40 mL acetonitriles, after mixing well, 20 min of ultrasonic extraction is filtered with filter paper to the tool plug added with 5 g sodium chloride
In graduated cylinder, 1 min is acutely vibrated, takes 20 mL supernatants to pour into 50 mL beakers after standing 1 h, beaker is placed in 40 DEG C
On electric boiling plate, keep its volatilization close dry, it is to be clean；
(4) sample purification: by step (3) volatilization nearly dry obtained residue 5 mL acetone, n-hexane and ethyl acetate
Mixed solution dissolution, amino solid-phase extraction column is then added, then it is molten with the mixing of 15 mL acetone, n-hexane and ethyl acetate
3 elutions of liquid point, merge leacheate, are placed on electric boiling plate and are evaporated in 40 DEG C, are settled to 2 mL with n-hexane, cross 0.22 μm of filter
Film obtains sample solution；
(5) sample detection: step (4) resulting sample solution is detected according to the chromatographic condition in step (2)；
(6) result calculates: according to formulaCalculate the concentration of cyflufenamid in sample；CIndicate to be measured
Cyflufenamid residual quantity in sample, mg/kg；C 0 Indicate concentration of standard solution, μ g/mL；AIndicate sample to be tested peak area；A 0 It indicates
Standard solution peak area；V 0 Indicate standard solution sampling volume, μ L；V 1 Indicate the volume of sample to be tested constant volume, mL；V 2 Indicate to
The sampling volume of sample, μ L；MIndicate the quality of sample to be tested, g.
Acetone in the mixed solution of acetone, n-hexane and ethyl acetate in the step (4), n-hexane and ethyl acetate
Volume ratio is 1: 1: 2.
Nh 2 column is eluted in advance with the mixed solution of 5 mL acetone, n-hexane and ethyl acetate in the step (4).
Beneficial effects of the present invention: 1, detection method of the invention is made using acetonitrile as extracting solution with reported acetone
Less compared to dosage for extracting solution, later purification is simpler；2, compared with methanol is as extracting solution, after extracting, saltouing, heating plate
It is evaporated, does not need to rotate, working efficiency can be improved；3, detection instrument popularity rate used is higher, and method easily promotes and applies；4, originally
The reproducibility of the detection method of invention, accuracy, precision and detection limit can meet retention analysis of the pesticide in wheat
It is required that；5, detection method of the invention reduces environmental pollution, reduces testing cost, improves work efficiency, and has stronger
Practicability has good application value.
Detailed description of the invention
Fig. 1 is 0.05 mg/L ring not bacterium amine standard solution chromatogram.
Fig. 2 is 0.1mg/L ring not bacterium amine standard solution chromatogram.
Fig. 3 is 0.5 mg/L ring not bacterium amine standard solution chromatogram.
Fig. 4 is 1 mg/L ring not bacterium amine standard solution chromatogram.
Fig. 5 is 2 mg/L rings not bacterium amine standard solution chromatogram.
Fig. 6 is the wheat flour control sample middle ring chromatogram that not bacterium amine additive amount is 0.02 mg/kg.
Fig. 7 is the wheat flour control sample middle ring chromatogram that not bacterium amine additive amount is 0.2 mg/kg.
Fig. 8 is the wheat flour control sample middle ring chromatogram that not bacterium amine additive amount is 1mg/kg.
Specific embodiment
Combined with specific embodiments below, the present invention will be further described.It should be understood that following embodiment is merely to illustrate this
The person skilled in the art of the range of invention and is not intended to limit the present invention, the field can make one according to the content of foregoing invention
A little nonessential modifications and adaptations.
The detection method of cyflufenamid residual quantity in wheat of the invention, the specific steps are as follows:
One, the content of cyflufenamid in wheat is measured
(1) preparation of standard solution: ring not bacterium amine standard items are weighed, are made 1000 mg/L's with n-hexane dissolution constant volume
Mother liquor, then it is diluted to the standard solution of 0.05mg/L, 0.1mg/L, 0.5mg/L, 1mg/L, 2mg/L step by step；
(2) draw standard curve: by standard solution made from step (1) cross chromatographic column RTX-5(30 m × 0.25 mm ×
0.25 μm) it is detected afterwards with ECD detector, injector temperature is 240 DEG C, and detector temperature is 300 DEG C, gas (N2) flow be 2
ML/min, make-up gas (N2) flow is 30mL/min, split ratio 30:1, sample volume is 1 μ L, is 180 DEG C of condition in column temperature
Then lower reservation 2min rises to 260 DEG C of 10 min of reservation according to the heating rate of 20 DEG C/min；
Using the concentration of standard solution as abscissa x, peak area is ordinate y, computer drawing standard curve, standard curve
Linear equation is y=175183.02x+17363.93, wherein x is the concentration of ring not bacterium amine standard solution, unit mg/L, y
For peak area, response related coefficient is r2=0.99；
(3) sample extraction: sample will be made after wheat crushing, weighs 10.00 g of the sample in 100 mL centrifugal bottles, adds
10 mL water and 40 mL acetonitriles, after mixing well, 20 min of ultrasonic extraction is filtered with filter paper to the tool plug added with 5 g sodium chloride
In graduated cylinder, 1 min is acutely vibrated, takes 20 mL supernatants to pour into 50 mL beakers after standing 1 h, beaker is placed in 40 DEG C
On electric boiling plate, keep its volatilization close dry, it is to be clean；
(4) sample purification: the residue that step (3) is evaporated 5 mL eluents (acetone: n-hexane: ethyl acetate=body
Product ratio 1: 1: 2) it in Yu Shangshu beaker dissolves, nh 2 column 5 mL elution liquid (acetone: n-hexane: ethyl acetate=volume
Than 1: 1: the eluent of dissolution residual substance in beaker is shifted upper prop by 2) pre- elution, with 15 mL eluents (acetone: n-hexane:
Ethyl acetate=volume ratio 1: 1: 2) being eluted, and is collected eluent, is placed on electric boiling plate and is evaporated for 40 DEG C, and n-hexane is accurately fixed
Hold to 2 mL, 0.22 μm of pin hole membrane filtration obtains sample solution；
The nearly dry obtained residue of step (3) volatilization is molten with the mixed solution of 5 mL acetone, n-hexane and ethyl acetate
Then solution is added amino solid-phase extraction column, then is eluted with mixed solution point 3 times of 15 mL acetone, n-hexane and ethyl acetate,
Merge leacheate, be placed on electric boiling plate and be evaporated in 40 DEG C, is settled to 2 mL with n-hexane, crosses 0.22 μm of filter membrane
(5) sample detection: by step (4) obtained sample solution cross chromatographic column RTX-5(30 m × 0.25 mm ×
0.25 μm) it is detected afterwards with ECD detector, injector temperature is 240 DEG C, and detector temperature is 300 DEG C, gas (N2) flow
For 2 mL/min, make-up gas (N2) it is 30 mL/min, split ratio 30:1, sample volume is 1 μ L, and column temperature is 180 DEG C
(2min), 20 DEG C/min are warming up to 260 DEG C (10 min)；
(6) result calculates: according to formulaCalculate the concentration of cyflufenamid in sample；CIt indicates to test sample
Cyflufenamid residual quantity in product, mg/kg；C 0 Indicate concentration of standard solution, μ g/mL；AIndicate sample to be tested peak area；A 0 Indicate mark
Quasi- solution peak area；V 0 Indicate standard solution sampling volume, μ L；V 1 Indicate the volume of sample to be tested constant volume, mL；V 2 Indicate to be measured
The sampling volume of sample, μ L；MIndicate the quality of sample to be tested, g.
Two, the TIANZHU XINGNAO Capsul of cyflufenamid in wheat is measured
Various concentration (0.02 mg/ is added in the wheat flour control sample without cyflufenamid prepared respectively
Kg, 0.2 mg/kg, 1 mg/kg) cyflufenamid standard specimen solution, shake up, place 2h after, according to above-mentioned steps (3) and step
(4) the sample extraction purification method described in is handled, and is detected to obtain standardized sample concentration by step (5), according to formulaTIANZHU XINGNAO Capsul is calculated, in formulaP For TIANZHU XINGNAO Capsul；C 1For sample solution concentration, i.e. sample is surveyed
Definite value (concentration of cyflufenamid in control sample)；C 2For standardized sample concentration, i.e. standardized sample measured value；C 3It is single for scalar quantity
Position is mg/kg.
The recovery of standard addition for measuring this method using the above method is as shown in table 1 below.
The rate of recovery and relative standard deviation of the cyflufenamid in wheat when 1 difference pitch-based sphere of table
As shown in Table 1, when pitch-based sphere is 0.02,0.2,1 mg/kg, average time of cyflufenamid in wheat is measured
Yield is 85~99%, and relative standard deviation is 0.77~5.3%, meets retention analysis requirement.
Claims (3)
1. the detection method of cyflufenamid residual quantity in a kind of wheat, it is characterised in that steps are as follows:
(1) preparation of standard solution: weighing cyflufenamid standard items, with n-hexane dissolution constant volume be made concentration be 0.05mg/L,
The standard solution of 0.1mg/L, 0.5mg/L, 1mg/L, 2mg/L；
(2) it draws standard curve: standard solution made from step (1) being crossed after chromatographic column RTX-5 and is detected with ECD detector, into
Sample mouth temperature is 240 DEG C, and detector temperature is 300 DEG C, and gas flow is 2 mL/min, and make-up gas flow is 30mL/min, point
For stream than being 30:1, sample volume is 1 μ L, retains 2min under conditions of column temperature is 180 DEG C, then according to the heating of 20 DEG C/min
Rate rises to 260 DEG C of 10 min of reservation；
Using the concentration of standard solution as abscissa x, peak area is ordinate y, and computer drawing standard curve, standard curve is linear
Equation is y=175183.02x+17363.93, wherein x is the concentration of cyflufenamid standard solution, and unit mg/L, y are peak
Area, response related coefficient are r2=0.99；
(3) sample extraction: sample will be made after wheat crushing, weighs the 10.00 g sample in 100 mL centrifugal bottles, adds 10
ML water and 40 mL acetonitriles, after mixing well, 20 min of ultrasonic extraction is filtered with filter paper to the tool plug graduated cylinder added with 5 g sodium chloride
In, 1 min is acutely vibrated, takes 20 mL supernatants to pour into 50 mL beakers after standing 1 h, the electricity that beaker is placed in 40 DEG C is added
On hot plate, keep its volatilization close dry, it is to be clean；
(4) sample purification: by the nearly dry obtained residue of step (3) volatilization mixing with 5 mL acetone, n-hexane and ethyl acetate
Solution dissolution is closed, amino solid-phase extraction column, then the mixed solution point 3 with 15 mL acetone, n-hexane and ethyl acetate is then added
Secondary elution merges leacheate, is placed on electric boiling plate and is evaporated in 40 DEG C, is settled to 2 mL with n-hexane, crosses 0.22 μm of filter membrane and obtains
To sample solution；
(5) sample detection: step (4) resulting sample solution is detected according to the chromatographic condition in step (2)；
(6) result calculates: according to formulaCalculate the concentration of cyflufenamid in sample；CIt indicates in sample to be tested
Cyflufenamid residual quantity, mg/kg；C 0 Indicate concentration of standard solution, μ g/mL；AIndicate sample to be tested peak area；A 0 Expression standard is molten
Liquid peak area；V 0 Indicate standard solution sampling volume, μ L；V 1 Indicate the volume of sample to be tested constant volume, mL；V 2 Indicate sample to be tested
Sampling volume, μ L；MIndicate the quality of sample to be tested, g.
2. the detection method of cyflufenamid residual quantity in wheat according to claim 1, it is characterised in that: the step
(4) volume ratio of acetone, n-hexane and ethyl acetate is 1: 1: 2 in the mixed solution of acetone, n-hexane and ethyl acetate in.
3. the detection method of cyflufenamid residual quantity in wheat according to claim 1, it is characterised in that: the step
(4) nh 2 column is eluted in advance with the mixed solution of 5 mL acetone, n-hexane and ethyl acetate in.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102914608A (en) * | 2012-11-12 | 2013-02-06 | 天津出入境检验检疫局动植物与食品检测中心 | Method for quickly detecting pesticide multiple residues in traditional Chinese medicine through membrane separation and comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometer |
| 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 |
| WO2015121802A1 (en) * | 2014-02-12 | 2015-08-20 | Isagro S.P.A. | Benzoylphenyl-formamidines having a fungicidal activity, their agronomic compositions and relative use |
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2016
- 2016-11-21 CN CN201611018989.4A patent/CN106770704B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102914608A (en) * | 2012-11-12 | 2013-02-06 | 天津出入境检验检疫局动植物与食品检测中心 | Method for quickly detecting pesticide multiple residues in traditional Chinese medicine through membrane separation and comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometer |
| 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 |
| WO2015121802A1 (en) * | 2014-02-12 | 2015-08-20 | Isagro S.P.A. | Benzoylphenyl-formamidines having a fungicidal activity, their agronomic compositions and relative use |
Non-Patent Citations (2)
| Title |
|---|
| 이윤정.Monitoring and Risk Assessment of Pesticide Residues in Vegetables among the Resident of Seoul, Korea.《Seoul National University Graduate School of Public Health》.2016,5-15. * |
| 固相萃取-气相色谱法对河水与海水中36种农药残留的同时测定;陈猛 等;《分析测试学报》;20091231;第28卷(第12期);5-15 * |
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Effective date of registration: 20210106 Address after: 463000 3rd floor, No.2 standard workshop, Southeast of the intersection of Xingye Avenue and Ruhe Avenue, Zhumadian City, Henan Province Patentee after: Henan LianJian pharmaceutical analysis Co.,Ltd. Address before: No.116 Huayuan Road, Jinshui District, Zhengzhou City, Henan Province Patentee before: INSTITUTE OF AGRICULTURAL QUALITY STANDARDS AND TESTING TECHNOLOGY, HENAN ACADEMY OF AGRICULTURAL SCIENCES |