CN115032300A - Method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide - Google Patents
Method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide Download PDFInfo
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Abstract
The invention relates to pesticide residue detection, in particular to a method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide, which comprises the following steps: extracting an animal source sample to be detected by using an extracting agent to obtain an extracting solution; performing EMR-Lipid purification column purification on the extracting solution to obtain a sample solution to be detected; and carrying out ultra performance liquid chromatography tandem mass spectrometry detection on the sample liquid to be detected to obtain detection results of fluopyram and 2- (trifluoromethyl) benzamide. The method provided by the invention can be used for quickly and accurately realizing qualitative and quantitative detection of fluopyram and 2- (trifluoromethyl) benzoyl in the sample to be detected.
Description
Technical Field
The invention relates to pesticide residue detection, in particular to a method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide.
Background
Fluopyram is a succinate dehydrogenase inhibitor (SDHIs) bactericide developed by Bayer GmbH, and has the main action mechanism of inhibiting electron transfer of succinate dehydrogenase in a respiratory chain and inhibiting mitochondrial respiration, so that spore germination, germ tube elongation, hypha growth and spore production of pathogenic bacteria are inhibited, and the purpose of controlling diseases is achieved. The fluopyram is mainly applied to crops such as tobacco, cucumber, tomato, banana, watermelon and the like, and has better control effects on various diseases such as powdery mildew, leaf spot, gray mold and the like and various nematodes.
Fluopyram is not easy to degrade in the environment, can easily enter water bodies through various ways such as permeation, surface runoff, atmospheric drift and the like after being applied in a large amount, causes pollution to the water environment, has higher toxicity to aquatic organisms, is easy to enrich in organisms, is transferred into organisms such as fish and the like through phytoplankton, is finally transferred into human beings through a food chain, and threatens human health.
According to the provisions of the joint conference of pesticide residues (JMPR), the residues of fluopyram in animal-derived samples are fluopyram precursors and 2- (trifluoromethyl) benzamide, a metabolite thereof. However, no report has been found for the simultaneous detection of fluopyram and the metabolite 2- (trifluoromethyl) benzamide.
Disclosure of Invention
The invention aims to provide a method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide, and the qualitative and quantitative detection of fluopyram and 2- (trifluoromethyl) benzoyl in a sample to be detected can be quickly and accurately realized by adopting the method provided by the invention.
In order to achieve the above object, the present invention provides a method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide, comprising the steps of:
extracting an animal source sample to be detected by using an extracting agent to obtain an extracting solution; performing EMR-Lipid purification column purification on the extracting solution to obtain a sample solution to be detected; performing ultra-high performance liquid chromatography tandem mass spectrometry detection on the sample liquid to be detected to obtain detection results of fluopyram and 2- (trifluoromethyl) benzamide;
the extractant comprises acetonitrile or acetonitrile formate solution.
The ultra-high performance liquid chromatography tandem mass spectrometry detection comprises ultra-high performance liquid chromatography detection and mass spectrometry detection; the conditions for detecting the ultra-high performance liquid chromatography comprise: the mobile phase system comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, and the mobile phase B is an ammonium acetate aqueous solution;
the flow rate of the mobile phase system is 300 mu L/min;
the elution mode is gradient elution;
the procedure for the gradient elution was:
0.0-0.5 min: the volume percentage content of the mobile phase A is 35 percent;
0.5-1.0 min: the volume percentage of the mobile phase A is increased from 35% to 80% at a constant speed;
1.0-2.5 min: the volume percentage content of the mobile phase A is 80%;
2.5-2.6 min: the volume percentage of the mobile phase A is reduced from 80% to 35% at a constant speed;
2.6-4.0 min: the volume percentage content of the mobile phase A is 35%;
the conditions for mass spectrometric detection include: an electrospray ion source; monitoring multiple reactions: an MRM mode; capillary voltage: 3.5 kV; nozzle voltage 500 ℃.
Preferably, the conditions of the ultra high performance liquid chromatography detection further comprise: the chromatographic column for the ultra-high performance liquid chromatography detection is Waters acquity
BEH C 18 A column; the column temperature was 40 ℃; the amount of sample was 1. mu.L.
Preferably, the mass spectrometric detection has a nebulizer pressure of 45 psi; the temperature of the drying gas is 300 ℃; the drying airflow rate is 5L/min; the temperature of the sheath gas is 250 ℃; the flow rate of the sheath gas is 11L/min; the nozzle voltage was 500 ℃.
Preferably, the concentration of the ammonium acetate aqueous solution is 2 mmol/L.
Preferably, the animal-derived sample to be tested comprises fish meat.
Preferably, the ratio of the mass of the animal-derived sample to the volume of the extractant is 2 g: 10 mL.
Preferably, the formic acid acetonitrile solution has a formic acid volume concentration of 0.1-2%.
The invention provides a method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide, which comprises the following steps: extracting an animal source sample to be detected by adopting an extracting agent to obtain an extracting solution; performing EMR-Lipid purification column purification on the extracting solution to obtain a sample solution to be detected; performing ultra-high performance liquid chromatography tandem mass spectrometry detection on the sample liquid to be detected to obtain detection results of fluopyram and 2- (trifluoromethyl) benzamide; the extractant comprises acetonitrile or a formic acid acetonitrile solution. The ultra-high performance liquid chromatography tandem mass spectrometry detection comprises ultra-high performance liquid chromatography detection and mass spectrometry detection; the conditions for detecting the ultra-high performance liquid chromatography comprise: the mobile phase system comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, and the mobile phase B is an ammonium acetate aqueous solution; the flow rate of the mobile phase system is 300 mu L/min; the elution mode is gradient elution; the procedure for the gradient elution was: 0.0-0.5 min: the volume percentage content of the mobile phase A is 35 percent; 0.5-1.0 min: the volume percentage of the mobile phase A is increased from 35% to 80% at a constant speed; 1.0-2.5 min: the volume percentage content of the mobile phase A is 80%; 2.5-2.6 min: the volume percentage of the mobile phase A is reduced from 80% to 35% at a constant speed; 2.6-4.0 min: the volume percentage content of the mobile phase A is 35 percent; the conditions for mass spectrometric detection include: an electrospray ion source; monitoring multiple reactions: an MRM mode; capillary voltage: 3.5 kV.
The method adopts acetonitrile or acetonitrile formate solution for extraction, and adopts EMR-Lipid purification column for purification to remove impurities such as protein, various lipids and the like in the animal-derived sample to be detected, so that the matrix effect is effectively reduced, the interference of the matrix is avoided, and meanwhile, the method is matched with the ultra-high performance liquid chromatography tandem mass spectrometry for detection, so that the sensitivity is high, the accuracy and the precision are high, and the method is suitable for simultaneously detecting the fluopyram and the 2- (trifluoromethyl) benzamide in the sample to be detected.
Drawings
FIG. 1 is a standard graph of fluopyram;
FIG. 2 is a standard graph of 2- (trifluoromethyl) benzamide;
FIG. 3 is a standard solution chromatogram of fluopyram (0.01 mg/L);
FIG. 4 is a chromatogram of a standard solution of 2- (trifluoromethyl) benzamide (0.01 mg/L).
Detailed Description
The invention provides a method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide, which comprises the following steps:
extracting an animal source sample to be detected by adopting an extracting agent to obtain an extracting solution;
performing EMR-Lipid purification column purification on the extracting solution to obtain a sample solution to be detected;
and carrying out ultra performance liquid chromatography tandem mass spectrometry detection on the sample liquid to be detected to obtain detection results of fluopyram and 2- (trifluoromethyl) benzamide.
In the present invention, the starting materials used in the present invention are preferably commercially available products unless otherwise specified.
The invention adopts an extracting agent to extract a sample to be detected of animal source to obtain an extracting solution.
In the present invention, the animal-derived sample to be tested preferably includes fish meat. In the present invention, the extractant comprises acetonitrile or a formic acid acetonitrile solution, preferably acetonitrile. In the invention, the formic acid acetonitrile solution preferably has a formic acid volume concentration of 0.1-2%. In the present invention, the ratio of the mass of the animal-derived sample to the volume of the extraction agent is preferably 2 g: 10 mL.
In the invention, the extraction mode is preferably extraction; the extraction preferably comprises: mixing an animal source sample to be detected, water and an extracting agent, and extracting to obtain an extraction system; and adding sodium chloride into the extraction system for centrifugation, and taking the supernatant as an extracting solution.
In the invention, the extraction is preferably carried out under the condition of vortex, and the time of vortex is preferably 10-15 min, and more preferably 10 min. In the invention, the rotation speed of the centrifugation is preferably 3000-4000 rpm, and more preferably 4000 rpm; the time of centrifugation is preferably 1-2 min.
After the extracting solution is obtained, the extracting solution is subjected to EMR-Lipid purification column purification to obtain the sample solution to be detected.
In the present invention, the EMR-Lipid purification column purification preferably comprises activating the EMR-Lipid purification column with water, and then adding the extract to the activated EMR-Lipid purification column to sequentially perform vortexing and centrifugation.
In the present invention, the vortexing preferably comprises an oscillatory vortexing; the time of the oscillating vortex is preferably 1 min; in the invention, the rotation speed of the centrifugation is preferably 4000-5000 r/min, more preferably 5000r/min, and the time of the centrifugation is preferably 5 min.
In the present invention, after purification, the present invention preferably further comprises subjecting the purified solution to EMR-Lipid Bond Elut Polish strip tube purification.
In the present invention, the EMR-Lipid Bond Elut Polish strip tube purification preferably comprises adding the solution obtained by the purification to an EMR-Lipid Bond Elut Polish strip tube, followed by vortexing and centrifugation.
In the present invention, the EMR-Lipid Bond Elut Polish strip tube purification can remove a small amount of moisture doped in the solution obtained by the purification.
In the present invention, the parameters and operation of the vortex and centrifugation are the same as the above-mentioned purification process, and are not described herein again.
After the sample liquid to be detected is obtained, the method carries out ultra performance liquid chromatography tandem mass spectrometry detection on the sample liquid to be detected to obtain the detection results of fluopyram and 2- (trifluoromethyl) benzamide.
In the invention, the ultra performance liquid chromatography tandem mass spectrometry detection comprises ultra performance liquid chromatography detection and mass spectrometry detection. In the invention, the conditions of the ultra-high performance liquid chromatography detection comprise:
the mobile phase system comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, and the mobile phase B is an ammonium acetate aqueous solution; the concentration of the ammonium acetate aqueous solution is 2mmol/L, and the flow rate of the mobile phase system is 300 mu L/min;
in the present invention, the elution mode is gradient elution. In the present invention, the procedure of the gradient elution is shown in table 1.
TABLE 1 liquid chromatography gradient elution conditions
| Gradient time/min | Mobile phase A/%) | Mobile phase B/%) |
| 0～0.5 | 35 | 65 |
| 0.5～1.0 | 80 | 20 |
| 1.0～2.5 | 80 | 20 |
| 2.5～2.6 | 35 | 65 |
| 2.6～4.0 | 35 | 65 |
In the present invention, the conditions for the liquid chromatography detection preferably further include: the chromatographic column is Waters acquity
BEH C 18 A column; the column temperature was 40 ℃; the amount of sample was 1. mu.L.
In the present invention, the electrospray ion source for mass spectrometry detection: ESI + (ii) a Monitoring multiple reactions: an MRM mode; capillary voltage: 3.5 KV.
In the invention, the atomizer pressure for mass spectrometric detection is 45 psi; the temperature of the drying gas is 300 ℃; the drying airflow rate is 5L/min; the temperature of the sheath gas is 250 ℃; the flow rate of the sheath gas is 11L/min; the nozzle voltage was 500 ℃.
In the present invention, the qualitative ion pair, the quantitative ion pair, and the collision energy of fluopyram and 2- (trifluoromethyl) benzamide are shown in table 2.
TABLE 2 Mass Spectrometry parameters for Fluopyram and 2- (trifluoromethyl) benzamide
Wherein denotes the quantitative ion.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It should be apparent that the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the embodiment of the present invention, the specifications of the specific instrument, reagent and sample to be tested are as follows:
agilent 6470B liquid chromatography tandem triple quadrupole mass spectrometer (Agilent Inc. USA), BSA224S electronic balance (sensitivity 0.0001g, Sidolis group, Germany);
an electronic balance of type OHAUS SPS202F (inductance 0.01g, Orxon instruments, Inc.);
eppendorf Centrifuge 5430R high speed refrigerated Centrifuge (Albendad, Germany);
MX-S adjustable mixing instrument (Beijing Dalongxing laboratory instruments Co., Ltd.);
acetonitrile (chromatographically pure 4L, Merck two-ply company);
formic acid (500 mL of chromatographic purity, Anaqua Chemicals Supply Co.);
sodium chloride (analytical purity 500g, Shanghai Lingfeng Chemicals Co., Ltd.);
cleanert MAS-Q purification tube (2mL, PSA 50mg, MgSO 4150 mg, C1850 mg, model MS-9PA0203, Tianjin Bonner Aijiel technologies, Inc.);
cleanert MAS-Q purifying tube (2mL, PSA 50mg, PC 8mg, C1850 mg, MgSO 4150 mg, model MS-9PP0265, Tianjin Bonnel technologies, Ltd.),
Cleanert MAS-Q purification tube (2mL, PSA 50mg, PC 50mg, C1850 mg, MgSO 4150 mg, model MS-9PP0250, Tianjin Bonner Egger science and technology Co., Ltd.);
EMR-Lipid SPE dispersed extraction tube (model 5982-;
EMR-Lipid Polish back extraction tube (model number 5982-;
an organic needle filter (0.22 μm, model TQP-61322, Tianyuan scientific Co.);
syringes (2mL, changzhouyuekang medical devices ltd);
the conditions of the ultra-high performance liquid chromatography detection are as follows: the chromatographic column is Waters acquity
BEH C 18 (1.7 μm, 2.1X 100 mm); the column temperature was 45 ℃; the sample injection amount is 1 mu L; the mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is methanol; the mobile phase B is 2mmol/l ammonium acetate aqueous solution; flow rate of the mobile phasePreferably 300. mu.L/min; the elution mode was gradient elution, and the conditions of gradient elution are shown in table 1.
The conditions for mass spectrometric detection include preferably: an ion source: ESI + (ii) a Quantitative mode: an MRM mode; capillary voltage: 3.5 kV; atomizer pressure: 45 psi; temperature of the drying gas: 300 ℃; flow rate of drying gas: 5L/min; temperature of sheath gas: 250 ℃; flow rate of sheath gas: 11L/min; nozzle voltage: at 500 ℃.
The qualitative ion pair, the quantitative ion pair and the collision energy of the fluopyram and the 2- (trifluoromethyl) benzamide are shown in the table 2.
Example 1
Extraction: and 4 groups of extraction tests are carried out on the sample to be detected by adopting different extracting agents, the extraction tests are respectively recorded as 1.1-1.4, and the extraction rate is calculated.
1.1 weighing 2g of fish sample, placing in a 50mL centrifuge tube, adding 2mL of water and 10mL of acetonitrile, vortexing for 10min, adding 2g of sodium chloride, vortexing for 1min, and centrifuging at 4000rpm for 5 min.
1.2 weighing 2g of fish meat sample, placing the fish meat sample in a 50mL centrifuge tube, adding 2mL of water and 10mL of 0.1% formic acid acetonitrile solution by volume, swirling for 10min, adding 2g of sodium chloride, swirling for 1min, and centrifuging for 5min at 4000 rpm.
1.3 weighing 2g of fish sample, placing the fish sample in a 50mL centrifuge tube, adding 2mL of water and 10mL of 1% formic acid acetonitrile solution by volume, swirling for 10min, adding 2g of sodium chloride, swirling for 1min, and centrifuging for 5min at 4000 rpm.
1.4 weighing 2g of fish meat sample, placing the fish meat sample in a 50mL centrifuge tube, adding 2mL of water and 10mL of 2% formic acid acetonitrile solution by volume, swirling for 10min, adding 2g of sodium chloride, swirling for 1min, and centrifuging for 5min at 4000 rpm.
Through detection, the extraction rates of fluopyram and 2- (trifluoromethyl) benzamide extracted by acetonitrile are 95.9% and 99.0% respectively; the extraction rates of fluopyram and 2- (trifluoromethyl) benzamide by using 0.1% volume concentration formic acid acetonitrile are respectively 100% and 96.9%. The extraction rates of extracting fluopyram and 2- (trifluoromethyl) benzamide by using 1% formic acid acetonitrile with volume concentration are respectively 99.1% and 98.0%; the extraction rates of fluopyram and 2- (trifluoromethyl) benzamide by extracting with 2% acetonitrile formate by volume concentration are 102% and 109%, respectively. The above data show that: the formic acid has no obvious influence on the extraction rate.
Example 2
Purifying: and 4 groups of purification tests are carried out on the sample to be detected by adopting different purifying agents, which are respectively recorded as 2.1-2.4, and the extraction rate is calculated.
2.1 taking 5mL of water-activated EMR-Lipid purification tube, uniformly mixing, adding 5mL of upper acetonitrile extracting solution, oscillating and whirling for 1min, centrifuging for 5min at 5000r/min, transferring the centrifuged supernatant into an EMR-Lipid Bond Elut Polish back-extraction tube, oscillating and whirling for 1min, centrifuging for 5min at 5000r/min, taking the supernatant, and passing the supernatant through a 0.22-micron organic needle filter to be determined.
2.2 accurately removing 1.5mL of the supernatant from the centrifuge tube, adding into MS-9PP0250 type Cleanert MAS-Q clean tube 1(2mL, PSA 50mg, PC 50mg, C1850 mg, MgSO 4150 mg), vortexing for 1min, centrifuging at 4000rpm for 5min, collecting the supernatant, passing through a 0.22 μm organic needle filter, and determining.
2.3 accurately removing 1.5mL of the supernatant from the centrifuge tube and adding into MS-9PP0265 type Cleanert MAS-Q purification tube 2(2mL, PSA 50mg, PC 8mg, C) 18 50mg、MgSO 4 150mg), vortexed for 1min, centrifuged at 4000rpm for 5min, and the supernatant was passed through a 0.22 μm organic needle filter and assayed.
2.4 accurately remove 1.5mL of the supernatant from the centrifuge tube and add into MS-9PA0203 type Cleanert MAS-Q purification tube 3(2mL, PSA 50mg, MgSO) 4 150mg、C 18 50mg), vortexed for 1min, centrifuged at 4000rpm for 5min, and the supernatant was passed through a 0.22 μm organic needle filter and assayed.
The results showed that the recovery rates of fluopyram and 2- (trifluoromethyl) benzamide were 71.6% and 97.3%, respectively, after purification by MS-9PP0250 type Cleanert MAS-Q purification tubes (2mL, PSA 50mg, PC 50mg, C1850 mg, MgSO 4150 mg), and the recovery rate of fluopyram was relatively low.
After being purified by an MS-9PP0265 type Cleanert MAS-Q purifying tube (2mL, PSA 50mg, PC 8mg, C1850 mg and MgSO 4150 mg), the recovery rates of fluopyram and 2- (trifluoromethyl) benzamide are respectively 107 percent and 116 percent, and the recovery rate of the 2- (trifluoromethyl) benzamide is relatively higher.
After purification by MS-9PA0203 type Cleanert MAS-Q purification tubes (2mL, PSA 50mg, MgSO 4150 mg, C1850 mg), the recovery rates of fluopyram and 2- (trifluoromethyl) benzamide were 106% and 115%, respectively, and the recovery rate of 2- (trifluoromethyl) benzamide was relatively high.
After EMR-Lipid purification, the recovery rates of fluopyram and 2- (trifluoromethyl) benzamide are respectively 105 percent and 103 percent, the recovery rates can meet the requirements, and impurities in the extracting solution are effectively removed. EMR-Lipid is an optimum purification condition in consideration of recovery rate, impurity removal effect, and the like, and thus, EMR-Lipid is determined as a purifier for fluopyram and 2- (trifluoromethyl) benzamide.
Example 3
First, standard curve:
preparing 0.0001, 0.0005, 0.001, 0.005, 0.01 and 0.05mg/L fluopyram and 2- (trifluoromethyl) benzamide standard solution by using a blank fish sample extracting solution, injecting 1 mu L of sample, detecting according to the conditions of ultra performance liquid chromatography detection and mass spectrometry detection to obtain a response value of UPLC-MS/MS, drawing a fluopyram standard solution curve by using concentration-peak area, and obtaining a result shown in figure 1 and a result shown in figure 2: the regression equation is that y is 3409791.83x-1268.51 (R) 2 0.9990); 2- (trifluoromethyl) benzamide standard solution curve, see fig. 2, with regression equation of y-2443519.43 x-928.88 (R) 2 0.9992). The fluopyram and the 2- (trifluoromethyl) benzamide of the invention have good linear relation in the concentration range of 0.0001-0.05 mg/L.
FIG. 3 is a standard solution chromatogram of fluopyram (0.01 mg/L); as can be seen from fig. 3: fluopyram peak shape is good, and ion pair quantification: 397.1>208.1, qualitative ion pair: 397.1>172.9, retention time is 2.868 min.
FIG. 4 is a chromatogram of a standard solution of 2- (trifluoromethyl) benzamide (0.01mg/L), and it can be seen from FIG. 4 that: the peak shape of the 2- (trifluoromethyl) benzamide is good, the quantitative ion pair is 190.1 to 170.1, the qualitative ion pair is 190.1 to 150.0, and the retention time is 1.961 min.
Second, measuring detection limit and quantitative limit
The method comprises the following steps: taking 0.01mg/L fluopyram and 2- (trifluoromethyl) benzamide standard solution to detect according to the conditions of ultra-high performance liquid chromatography detection and mass spectrometry detection, determining the quantitative limit according to the corresponding concentration of the ratio of a detected component signal (S) to baseline noise (N) being more than or equal to 10(S/N being more than or equal to 10), and determining the detection limit according to the corresponding concentration of S/N being more than or equal to 3.
The results were: the detection limits of fluopyram and 2- (trifluoromethyl) benzamide are both 0.01 mg/kg.
Instrument detection limit: the minimum detection amount of ultra-high performance liquid chromatography tandem mass spectrometer (UPLC-MS/MS) to fluopyram and 2- (trifluoromethyl) benzamide is 1 × 10 -13 g。
Example 4
Determination of recovery
Adding appropriate amount of fluopyram and 2- (trifluoromethyl) benzamide standard substance into the blank fish meat respectively to make the addition concentration of fluopyram and 2- (trifluoromethyl) benzamide standard substance be 0.01, 0.1 and 5.0mg/kg respectively, and setting the addition concentration of each grade to be 5 times.
Then extracting according to 1.1 in example 1 and purifying according to 2.1 in example 2 to obtain a sample liquid to be detected, and detecting according to the detection condition of ultra performance liquid chromatography tandem mass spectrometry to obtain the addition recovery rate and Relative Standard Deviation (RSDs) of fluopyram and 2- (trifluoromethyl) benzamide in the sample to be detected, and the RSDs are shown in Table 3.
TABLE 3 recovery and relative standard deviation of Fluopyram and 2- (trifluoromethyl) benzamide additions
As can be seen from table 3, when fluopyram is added to fish meat at concentrations of 0.01, 0.1 and 5.0mg/kg, the average recovery rates are 96.2%, 90.2% and 96.0%, respectively, and the relative standard deviations are 9.7%, 2.3% and 9.8%, respectively; 2- (trifluoromethyl) benzamide was added to fish meat at concentrations of 0.01, 0.1 and 5.0mg/kg, with average recoveries of 89.3%, 88.7% and 103%, respectively, and relative standard deviations of 3.5%, 1.6% and 7.7%, respectively.
In summary, the following steps: the detection method provided by the invention has the advantages that the average recovery rate of fluopyram in fish meat is 82.0-113%, and the relative standard deviation is 2.3-9.8%; the average recovery rate of the 2- (trifluoromethyl) benzamide in the fish is 84.8-111 percent, and the relative standard deviation is 1.6-7.7 percent. The minimum detection concentrations of fluopyram and 2- (trifluoromethyl) benzamide are both 0.01 mg/kg. The method is quick, sensitive, simple, convenient, efficient and good in purification effect, and is suitable for quickly detecting the residue of fluopyram and 2- (trifluoromethyl) benzamide.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (7)
1. A method for simultaneously detecting fluopyram and 2- (trifluoromethyl) benzamide comprises the following steps:
extracting an animal source sample to be detected by using an extracting agent to obtain an extracting solution;
performing EMR-Lipid purification column purification on the extracting solution to obtain a sample solution to be detected;
performing ultra-high performance liquid chromatography tandem mass spectrometry detection on the sample liquid to be detected to obtain detection results of fluopyram and 2- (trifluoromethyl) benzamide;
the extractant comprises acetonitrile or a formic acid acetonitrile solution;
the ultra-high performance liquid chromatography tandem mass spectrometry detection comprises ultra-high performance liquid chromatography detection and mass spectrometry detection; the conditions for detecting the ultra-high performance liquid chromatography comprise: the mobile phase system comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, and the mobile phase B is an ammonium acetate aqueous solution;
the flow rate of the mobile phase system is 300 mu L/min;
the elution mode is gradient elution;
the procedure for the gradient elution was:
0.0-0.5 min: the volume percentage content of the mobile phase A is 35 percent;
0.5-1.0 min: the volume percentage of the mobile phase A is increased from 35% to 80% at a constant speed;
1.0-2.5 min: the volume percentage content of the mobile phase A is 80%;
2.5-2.6 min: the volume percentage of the mobile phase A is reduced from 80% to 35% at a constant speed;
2.6-4.0 min: the volume percentage content of the mobile phase A is 35 percent;
the conditions for mass spectrometric detection comprise: an electrospray ion source; monitoring multiple reactions: an MRM mode; capillary voltage: 3.5 kV.
2. The method as claimed in claim 1, wherein the HPLC detection column is a Waters acquity
BEH C 18 A column; the column temperature was 40 ℃; the amount of sample was 1. mu.L.
3. The method of claim 1, wherein the mass spectrometric detection has a nebulizer pressure of 45 psi; the temperature of the drying gas is 300 ℃; the drying airflow rate is 5L/min; the temperature of the sheath gas is 250 ℃; the flow rate of the sheath gas is 11L/min; the nozzle voltage was 500 ℃.
4. The method according to claim 1, wherein the concentration of the aqueous ammonium acetate solution is 2 mmol/L.
5. The method of claim 1, wherein the test sample of animal origin comprises fish.
6. The method according to claim 1, wherein the ratio of the mass of the animal-derived test sample to the volume of the extractant is 2 g: 10 mL.
7. The method according to claim 1, wherein the formic acid acetonitrile solution has a formic acid volume concentration of 0.1-2%.
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| CN104730191A (en) * | 2015-03-12 | 2015-06-24 | 崔淑华 | LC-MS/MS determination method of residual quantity of fluoroether bacteria amide |
| CN108896694A (en) * | 2018-07-05 | 2018-11-27 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of remaining LC-QToF-MS Screening analysis method of pesticide in animal food |
| WO2022077808A1 (en) * | 2020-10-16 | 2022-04-21 | 宁波市疾病预防控制中心 | Method and kit for measuring fentanyl drug in biological sample |
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|---|---|---|---|---|
| CN104730191A (en) * | 2015-03-12 | 2015-06-24 | 崔淑华 | LC-MS/MS determination method of residual quantity of fluoroether bacteria amide |
| CN108896694A (en) * | 2018-07-05 | 2018-11-27 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of remaining LC-QToF-MS Screening analysis method of pesticide in animal food |
| WO2022077808A1 (en) * | 2020-10-16 | 2022-04-21 | 宁波市疾病预防控制中心 | Method and kit for measuring fentanyl drug in biological sample |
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