CN111965274A - Method for analyzing and detecting clopyralid in rapeseeds - Google Patents
Method for analyzing and detecting clopyralid in rapeseeds Download PDFInfo
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- CN111965274A CN111965274A CN202010775529.6A CN202010775529A CN111965274A CN 111965274 A CN111965274 A CN 111965274A CN 202010775529 A CN202010775529 A CN 202010775529A CN 111965274 A CN111965274 A CN 111965274A
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- 240000002791 Brassica napus Species 0.000 title claims abstract description 61
- 239000005500 Clopyralid Substances 0.000 title claims abstract description 59
- HUBANNPOLNYSAD-UHFFFAOYSA-N clopyralid Chemical compound OC(=O)C1=NC(Cl)=CC=C1Cl HUBANNPOLNYSAD-UHFFFAOYSA-N 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 26
- 235000011293 Brassica napus Nutrition 0.000 title claims abstract description 25
- 235000004977 Brassica sinapistrum Nutrition 0.000 claims abstract description 50
- 239000011159 matrix material Substances 0.000 claims abstract description 26
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 18
- 238000001514 detection method Methods 0.000 claims abstract description 18
- 238000004458 analytical method Methods 0.000 claims abstract description 8
- 239000007864 aqueous solution Substances 0.000 claims abstract description 8
- 238000000589 high-performance liquid chromatography-mass spectrometry Methods 0.000 claims abstract description 4
- 244000188595 Brassica sinapistrum Species 0.000 claims abstract 14
- 239000000243 solution Substances 0.000 claims description 26
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 239000012224 working solution Substances 0.000 claims description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 239000007789 gas Substances 0.000 claims description 9
- 238000012417 linear regression Methods 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 8
- 238000005303 weighing Methods 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 7
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 6
- 238000012544 monitoring process Methods 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 238000001819 mass spectrum Methods 0.000 claims description 5
- 238000007664 blowing Methods 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- 238000002552 multiple reaction monitoring Methods 0.000 claims description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 3
- 229910052786 argon Inorganic materials 0.000 claims description 3
- 238000000889 atomisation Methods 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 235000019253 formic acid Nutrition 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 238000011895 specific detection Methods 0.000 claims description 2
- 238000011084 recovery Methods 0.000 abstract description 13
- 238000012360 testing method Methods 0.000 abstract description 8
- 239000012535 impurity Substances 0.000 abstract description 3
- 239000000523 sample Substances 0.000 description 22
- 239000000284 extract Substances 0.000 description 16
- 241000196324 Embryophyta Species 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 239000011550 stock solution Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 238000004364 calculation method Methods 0.000 description 4
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- 238000000605 extraction Methods 0.000 description 4
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- 239000012491 analyte Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
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- 238000000861 blow drying Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
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- 230000014509 gene expression Effects 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
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Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
Abstract
The invention discloses an analysis and detection method of clopyralid in rapeseeds, which adopts a high performance liquid chromatography-mass spectrometry combined method for detection, uses 2% of sodium hydroxide aqueous solution to extract a sample to be detected, and verifies the effectiveness and accuracy of the method by adding a recovery test. When the addition range of the clopyralid in the rapeseeds is 0.05-4 mg/kg, the recovery rate is 98-104%, and the Relative Standard Deviation (RSD) is 0.7-2.8%. The method solves the problem of large impurity interference in oil crops, and can efficiently and accurately detect the content of clopyralid in a rapeseed matrix.
Description
Technical Field
The invention relates to the technical field of pesticide residue detection, in particular to an analysis and detection method of clopyralid in rapeseeds.
Background
The clopyralid is a synthetic plant growth hormone, is absorbed by roots and leaves of plants and then conducted in the bodies of the plants, can effectively stimulate the synthesis of DNA, RNA and protein of the plants so as to cause the uncontrolled and disordered growth of cell division and finally cause the destruction of tube bundles, and is widely used for preventing and killing annual broad leaf weeds and perennial broad leaf weeds with deep roots in fields such as rapes, corns, lawns and the like.
The prior literature reports that clopyralid is detected by liquid chromatography, gas chromatography and liquid chromatography-mass spectrometry. For oil crops, the existing liquid chromatography and gas chromatography have higher limit of quantitation, can not detect samples with less residual quantity, and can not meet the requirement of trace analysis; the existing liquid chromatography-mass spectrometry method is poor in applicability, impurities in oil crops cannot be effectively removed in the pretreatment process, the recovery rate cannot meet the requirement, and research is carried out on the basis of the requirement to find a reliable detection method for detecting clopyralid in rapeseeds.
Disclosure of Invention
Based on the technical problems in the background art, the invention provides an analysis and detection method of clopyralid in rapeseeds, and the invention can quickly, conveniently and accurately detect the content of clopyralid in a rapeseed matrix.
The invention provides an analysis and detection method of clopyralid in rapeseeds, which adopts a high performance liquid chromatography-mass spectrometry combined method for detection, wherein the conditions of the high performance liquid chromatography are as follows: the chromatographic column is a Shim-pack XR-ODS II chromatographic column, the mobile phase A is formic acid aqueous solution with the volume fraction of 0.1 percent, the mobile phase B is acetonitrile, the flow rate is 0.2mL/min, gradient elution is carried out,
the gradient elution procedure was: within 0-0.5min, the proportion of the mobile phase A is 90 percent, and the proportion of the mobile phase B is 10 percent; the proportion of the mobile phase A is gradually changed from 90% to 40% within 0.5-1.2 min; within 1.2-2.0min, the proportion of the mobile phase A is gradually changed from 40% to 10%; the proportion of the mobile phase A is maintained at 10 percent within 2.0-2.5 min; within 2.5-2.51min, the proportion of the mobile phase A is gradually changed from 10% to 90%; the proportion of the mobile phase A is maintained at 90 percent within 2.51-5 min;
the mass spectrum conditions are as follows: the ion source is an electrospray ion source under atmospheric pressure, in a negative ion mode, and is a triple quadrupole mass analyzer, the interface voltage is 3.5kv, the DL tube temperature is 250 ℃, the heating block temperature is 400 ℃, the atomization gas flow is 3L/min, the drying gas flow is 15L/min, and the collision gas is argon; the monitoring mode is a multi-reaction monitoring mode.
Preferably, the multiple reaction monitoring conditions for clopyralid are: the mass-to-charge ratio of the qualitative ion pair is 190.00>146.00 and 192.00>147.95, and the mass-to-charge ratio of the quantitative ion pair is 190.00> 146.00; wherein, the deviation voltage of Q1pre, the deviation voltage of collision voltage CE and Q3pre corresponding to ion pair 190.00>146.00 are respectively 21V, 12 and 27V, the deviation voltage of Q1pre, the deviation voltage of collision voltage CE and Q3pre corresponding to ion pair 192.00>147.95 are respectively 21V, 9 and 27V, and the residence time is 100 msec.
The symbol ">" in the above-mentioned ion pair is a symbol commonly used by those skilled in the art to represent the ion pair.
The Q1pre deviation voltage, the collision voltage CE, and the Q3pre deviation voltage are specific expressions of the liquid chromatography-mass spectrometer of shimadzu corporation, japan.
Preferably, the size of the Shim-pack XR-ODS II column is 2.0 mm.d.times.75 mm.
Preferably, the column temperature is 40 ℃.
Preferably, the sample size is 2 μ L.
Preferably, the specific detection steps are: taking a clopyralid standard substance, preparing series standard working solutions with different concentrations by using a rapeseed blank matrix extracting solution, injecting a sample and drawing a standard curve to obtain a linear regression equation, then taking a rapeseed extracting solution to be detected for injection, and calculating the content of the clopyralid in the rapeseeds by using the linear regression equation.
Preferably, rapeseed blank matrix refers to rapeseed matrix without clopyralid.
Preferably, the preparation method of the rapeseed blank matrix extracting solution is the same as that of the rapeseed extracting solution to be detected, and the preparation method comprises the following steps: weighing 5.000g of crushed and uniformly mixed rapeseed blank matrix or rapeseed to be detected, whirling with 15mL of 2% sodium hydroxide aqueous solution for 5min, adjusting the pH value to 2-3 with concentrated hydrochloric acid, then adding 10mL of ethyl acetate, whirling, mixing uniformly, centrifuging, transferring 2mL of supernatant, drying by using nitrogen, dissolving the residual residue after drying by blowing by using 1mL of methanol, centrifuging, and passing the supernatant through a 0.22 mu m organic filter membrane.
To prove the effectiveness of the verification method, a clopyralid addition recovery test in rapeseeds was carried out for this purpose, and the results are as follows:
when the addition concentration of the clopyralid in the rapeseeds is 0.05-4 mg/kg, the recovery rate is 98-104%, the Relative Standard Deviation (RSD) is 0.7-2.8%, and the recovery rate and the relative standard deviation meet the requirement of NY/T788-2018; the minimum detection amount of the clopyralid in the rapeseeds is 40pg, the quantitative limit of the clopyralid is 0.05mg/kg, and the maximum residual limit requirement at home and abroad is met (GB 2763-2019 provides that the maximum residual limit value of the clopyralid in the rapeseeds is 0.05 mg/kg).
Has the advantages that:
the method uses an LC-MS/MS (high performance liquid chromatography-mass spectrometry) analysis technology to qualitatively determine the substances to be detected through dual conditions of retention time and ion abundance ratio; by changing the extraction and purification method and optimizing the detection conditions of the instrument, the sensitivity and the resolution capability of the instrument are improved, the problem of high impurity interference in rapeseed matrixes is solved, the limit of quantitative limit is broken through, and the detection of the residual quantity of the clopyralid in trace level can be met. The invention can efficiently and accurately detect the content of clopyralid in the rapeseed matrix.
Drawings
FIG. 1 is a standard curve of clopyralid in a standard working solution of example 1.
FIG. 2 is an extracted ion current chromatogram of the white solvent in example 1.
FIG. 3 is an extracted ion current chromatogram of the extract solution of rapeseed blank matrix in example 1.
FIG. 4 is an extracted ion current chromatogram of clopyralid in the standard working solution in example 1.
FIG. 5 is a chromatogram of the extracted ion current of the rapeseed extract to be tested in example 1.
FIG. 6 is an extracted ion current chromatogram of an extract solution of sample A in example 2.
FIG. 7 is an extracted ion current chromatogram of an extract solution of sample B in example 2.
FIG. 8 is an extracted ion current chromatogram of an extract solution of sample C in example 2.
FIG. 9 is an extracted ion current chromatogram of an extract solution of sample D in example 2.
FIG. 10 is an extracted ion current chromatogram of an extract liquid of sample E in example 2.
FIG. 11 is an extracted ion current chromatogram of an extract solution of sample F in example 2.
Detailed Description
The technical solution of the present invention will be described in detail below with reference to specific examples.
The main apparatus comprises:
liquid chromatography-mass spectrometer (LCMS-8040), shimadzu corporation, japan;
one-tenth-of-ten-thousandth electronic balance (AUW-220D), Shimadzu corporation, Japan;
vortex mixer (XH-D), Shanghai Hano instruments, Inc.;
water bath constant temperature oscillator (GY2016-SW), manufactured by yoyo instruments ltd, japan;
centrifuge (TDZ5-WS), Hunan instrument laboratory Instrument development Co., Ltd.
The main reagents are as follows:
clopyralid standard (purity 98.1%); methanol (chromatographically pure); acetonitrile (chromatographically pure); ethyl acetate (chromatographically pure); sodium hydroxide (analytical grade); hydrochloric acid (analytically pure); pure water (primary water).
Example 1
An analysis and detection method of clopyralid in rapeseeds adopts a liquid chromatography tandem mass spectrometer LCMS-8050 to detect the clopyralid in Shimadzu corporation, wherein the conditions of high performance liquid chromatography are as follows: the chromatographic column is a Shim-pack XR-ODS II chromatographic column (2.0 mm. d. times.75 mm), the mobile phase A is 0.1% formic acid aqueous solution by volume fraction, the mobile phase B is acetonitrile, the flow rate is 0.2mL/min, the column temperature is 40 ℃, the sample injection amount is 2 μ L, gradient elution is carried out,
the gradient elution procedure was: within 0-0.5min, the proportion of the mobile phase A is 90 percent, and the proportion of the mobile phase B is 10 percent; the proportion of the mobile phase A is gradually changed from 90% to 40% within 0.5-1.2 min; within 1.2-2.0min, the proportion of the mobile phase A is gradually changed from 40% to 10%; the proportion of the mobile phase A is maintained at 10 percent within 2.0-2.5 min; within 2.5-2.51min, the proportion of the mobile phase A is gradually changed from 10% to 90%; the proportion of the mobile phase A is maintained at 90 percent within 2.51-5 min;
the mass spectrum conditions are as follows: the ion source is an electrospray ion source under atmospheric pressure, in a negative ion mode, and is a triple quadrupole mass analyzer, the interface voltage is 3.5kv, the DL tube temperature is 250 ℃, the heating block temperature is 400 ℃, the atomization gas flow is 3L/min, the drying gas flow is 15L/min, and the collision gas is argon; the monitoring mode is a multi-reaction monitoring mode;
the multiple reaction monitoring conditions for clopyralid are shown in table 1:
TABLE 1 multiple reaction monitoring conditions for clopyralid
Note: plus ". sup." indicates the quantitative ion.
Solution preparation:
blank solution: methanol.
The pretreatment process of the rapeseed blank matrix is the same as that of the rapeseed to be tested.
Sample preparation: taking blank rapeseeds or rapeseeds to be detected, uniformly mixing the blank rapeseeds or the rapeseeds to be detected in a stainless steel basin, dividing the mixture by a quartering method, uniformly crushing the divided sample by a crusher, and storing the sample in a sealed way to prepare marks for later use.
Rapeseed blank matrix extract: weighing 5.001g of crushed and uniformly mixed rapeseed blank matrix into a 50mL centrifuge tube, adding 15mL of sodium hydroxide aqueous solution with the mass fraction of 2%, performing vortex extraction for 5min, adjusting the pH value to be 2-3 by using concentrated hydrochloric acid, performing vortex mixing for 3min, adding 10mL of ethyl acetate, performing vortex mixing for 5min, centrifuging for 5min at 5000r/min, accurately transferring 2mL of supernatant into a 10mL centrifuge tube, performing nitrogen blowing on the supernatant to dryness by using a water bath nitrogen blower, dissolving and blow-drying the residue remained after drying by using 1mL of chromatographic methanol, performing vortex mixing for 1min, performing centrifugation for 1min at 3000r/min, and filtering the supernatant through a 0.22 mu m organic filter membrane to obtain the rapeseed solution to be detected.
Rapeseed solution to be tested: weighing 5.003g of crushed and uniformly-mixed rapeseeds to be detected in a 50mL centrifuge tube, adding 15mL of a 2% sodium hydroxide aqueous solution by mass fraction, performing vortex extraction for 5min, adjusting the pH value to be 2-3 by using concentrated hydrochloric acid, performing vortex mixing for 3min, adding 10mL of ethyl acetate, performing vortex mixing for 5min, performing centrifugation for 5min at 5000r/min, accurately transferring 2mL of supernate into a 10mL centrifuge tube, performing nitrogen blowing on the supernate to dryness by using a water bath nitrogen blower, dissolving and drying the residual residue after drying by using 1mL of chromatographic methanol, performing vortex mixing for 1min, performing centrifugation for 1min at 3000r/min, and filtering the supernate through a 0.22 mu m organic filter membrane to obtain a rapeseeds solution to be detected.
Standard working solution: weighing (accurate to 0.00001g) proper amount of clopyralid standard substance, dissolving with chromatographic pure acetonitrile, and preparing into standard stock solution with concentration of 1000 mg/L; then precisely transferring a proper amount of standard stock solution into a volumetric flask, diluting with the rapeseed blank matrix extracting solution, fixing the volume, and preparing into series of standard working solutions with different concentrations.
The operation method comprises the following steps: setting instrument parameters according to the chromatographic and mass spectrum conditions, editing a batch processing table after the instrument is stabilized, and sequentially collecting a reagent blank solvent, a rapeseed blank matrix extracting solution, a series of standard working solutions and a rapeseed extracting solution to be detected; analyzing the collected data, drawing a standard curve to obtain a linear regression equation, and calculating the content of clopyralid in the rapeseeds to be detected according to the linear regression equation by an external standard method.
FIG. 1 is a standard curve of clopyralid standard substance in example 1, wherein the abscissa is the concentration X of clopyralid standard substance and the ordinate is the peak area f (X) of clopyralid standard substance, and the linear regression equation obtained is f (X) -242428X +1328.69, R20.9998023; the table on the right in FIG. 1 shows the peak areas corresponding to different concentrations (mg/L) of clopyralid in the standard working solution.
The calculation formula of the residual amount of clopyralid in the rapeseeds to be detected is as follows:
Xtest object＝CTest object×V0/mRapeseed
In the formula:
Xtest object-the residual amount of the test substance in mg/kg in rapeseed;
Ctest object-the concentration of the analyte in mg/L in the rapeseed extract;
V0-the volume of extraction reagent added, L, when preparing rapeseed extract;
mrapeseedWeighing the weight of the rapeseed to be tested, kg.
The content of clopyralid in rapeseed was found to be 2.03 mg/kg.
Typical extracted ion flow chromatograms are shown in fig. 2-5.
FIG. 2 is an extracted ion current chromatogram of the white solvent in example 1.
FIG. 3 is an extracted ion current chromatogram of the extract solution of rapeseed blank matrix in example 1.
FIG. 4 is an extracted ion current chromatogram of the standard working solution of example 1, wherein the retention time of clopyralid is 3.945 min.
FIG. 5 is the chromatogram of the extracted ion current of the rapeseed extract to be tested in example 1, wherein the retention time of clopyralid is 3.927 min.
Example 2 recovery test
The residual amount of clopyralid in the sample was measured by adding a sample of known concentration according to the method and detection conditions of example 1, and the recovery rate was calculated.
The experimental process comprises the following steps:
weighing 6 parts of rapeseed blank matrix, weighing 2.000g of rapeseed blank matrix with the number of A, B, C, D, E, F in each part, then respectively diluting clopyralid standard stock solutions (same as example 1) to different concentrations, adding the clopyralid standard stock solutions into A, B, C, D, E, F rapeseed blank matrix, and uniformly mixing the clopyralid standard stock solutions to ensure that the clopyralid addition concentration in a A, B sample is 0.05 mg/kg; C. d, the addition concentration of the clopyralid in the sample is 2 mg/kg; E. the addition concentration of the clopyralid in the sample F is 4 mg/kg; then standing the sample for 2 h;
the extracts of A, B, C, D, E, F samples were obtained by treating the rapeseed extracts to be tested in example 1.
The blank solvent, rapeseed blank matrix extract and standard working solution were the same as in example 1.
The operation method comprises the following steps: setting instrument parameters according to the chromatographic and mass spectrum conditions, editing a batch processing table after the instrument is stabilized, and sequentially collecting a reagent blank solvent, a rapeseed blank matrix extracting solution, a series of standard working solutions and an A, B, C, D, E, F extracting solution; analyzing the collected data, drawing a standard curve to obtain a linear regression equation, and calculating A, B, C, D, E, F the content of clopyralid in the sample according to the linear regression equation by an external standard method and calculating the recovery rate of the sample.
The sample recovery rate calculation formula is as follows:
in the formula: x is recovery (%); c1The detection value of pesticide added in the rapeseed blank matrix sample is mg/kg; c0The concentration value of the actually added pesticide in the blank sample is mg/kg.
The results of the recovery calculation are shown in Table 2.
TABLE 2 results of recovery calculation
Typical chromatograms are shown in FIGS. 6-11.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (8)
1. The method for analyzing and detecting the clopyralid in the rapeseeds is characterized by adopting a high performance liquid chromatography-mass spectrometry combined method for detection, wherein the conditions of the high performance liquid chromatography are as follows: the chromatographic column is a Shim-pack XR-ODS II chromatographic column, the mobile phase A is formic acid aqueous solution with the volume fraction of 0.1 percent, the mobile phase B is acetonitrile, the flow rate is 0.2mL/min, gradient elution is carried out,
the gradient elution procedure was: within 0-0.5min, the proportion of the mobile phase A is 90 percent, and the proportion of the mobile phase B is 10 percent; the proportion of the mobile phase A is gradually changed from 90% to 40% within 0.5-1.2 min; within 1.2-2.0min, the proportion of the mobile phase A is gradually changed from 40% to 10%; the proportion of the mobile phase A is maintained at 10 percent within 2.0-2.5 min; within 2.5-2.51min, the proportion of the mobile phase A is gradually changed from 10% to 90%; the proportion of the mobile phase A is maintained at 90 percent within 2.51-5 min;
the mass spectrum conditions are as follows: the ion source is an electrospray ion source under atmospheric pressure, in a negative ion mode, and is a triple quadrupole mass analyzer, the interface voltage is 3.5kv, the DL tube temperature is 250 ℃, the heating block temperature is 400 ℃, the atomization gas flow is 3L/min, the drying gas flow is 15L/min, and the collision gas is argon; the monitoring mode is a multi-reaction monitoring mode.
2. The method for analyzing and detecting clopyralid in rapeseed as claimed in claim 1, wherein the multiple reaction monitoring conditions of clopyralid are as follows: the mass-to-charge ratio of the qualitative ion pair is 190.00>146.00 and 192.00>147.95, and the mass-to-charge ratio of the quantitative ion pair is 190.00> 146.00; wherein, the deviation voltage of Q1pre, the deviation voltage of collision voltage CE and Q3pre corresponding to ion pair 190.00>146.00 are respectively 21V, 12 and 27V, the deviation voltage of Q1pre, the deviation voltage of collision voltage CE and Q3pre corresponding to ion pair 192.00>147.95 are respectively 21V, 9 and 27V, and the residence time is 100 msec.
3. The analytical detection method for clopyralid in rapeseed as claimed in claim 1 or 2, characterized in that the size of a Shim-pack XR-ODS II chromatographic column is 2.0 mm.d.times.75 mm.
4. The analytical method for the detection of clopyralid in rapeseed as claimed in any of claims 1 to 3, characterized in that the column temperature is 40 ℃.
5. The method for the analytical detection of clopyralid in rapeseed as claimed in any one of claims 1 to 4, characterized in that the sample volume is 2 μ L.
6. The analytical method for clopyralid in rapeseed according to any one of claims 1 to 5, characterized in that the specific detection steps are as follows: taking a clopyralid standard substance, preparing series standard working solutions with different concentrations by using a rapeseed blank matrix extracting solution, injecting a sample and drawing a standard curve to obtain a linear regression equation, then taking a rapeseed extracting solution to be detected for injection, and calculating the content of the clopyralid in the rapeseeds by using the linear regression equation.
7. The method of claim 6, wherein the rapeseed blank matrix is rapeseed matrix without clopyralid.
8. The method for analyzing and detecting clopyralid in rapeseeds according to claim 6, wherein the preparation method of the rapeseed blank matrix extracting solution is the same as that of the rapeseed extracting solution to be detected, and the preparation method comprises the following steps: weighing 5.000g of crushed and uniformly mixed rapeseed blank matrix or rapeseed to be detected, whirling with 15mL of 2% sodium hydroxide aqueous solution for 5min, adjusting the pH value to 2-3 with concentrated hydrochloric acid, then adding 10mL of ethyl acetate, whirling, mixing uniformly, centrifuging, transferring 2mL of supernatant, drying by using nitrogen, dissolving the residual residue after drying by blowing by using 1mL of methanol, centrifuging, and passing the supernatant through a 0.22 mu m organic filter membrane.
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