CN114384157A - Method for detecting residual quantity of imazethapyr in milk or bean beverage - Google Patents
Method for detecting residual quantity of imazethapyr in milk or bean beverage Download PDFInfo
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- XVOKUMIPKHGGTN-UHFFFAOYSA-N Imazethapyr Chemical compound OC(=O)C1=CC(CC)=CN=C1C1=NC(C)(C(C)C)C(=O)N1 XVOKUMIPKHGGTN-UHFFFAOYSA-N 0.000 title claims abstract description 18
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- NUPJIGQFXCQJBK-UHFFFAOYSA-N 2-(4-isopropyl-4-methyl-5-oxo-4,5-dihydro-1H-imidazol-2-yl)-5-(methoxymethyl)nicotinic acid Chemical compound OC(=O)C1=CC(COC)=CN=C1C1=NC(C)(C(C)C)C(=O)N1 NUPJIGQFXCQJBK-UHFFFAOYSA-N 0.000 description 1
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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
-
- 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
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- 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)
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- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention provides a method for detecting residual quantity of imazethapyr in milk or bean beverage, which comprises the following steps: mixing a sample to be detected with an acidified first solvent to obtain a mixture so as to extract residual imazapic in the sample to be detected; adding sodium chloride into the mixture, uniformly mixing, centrifuging, and collecting supernatant; carrying out solid phase extraction on the supernatant, and collecting all effluent liquid; drying the effluent, redissolving the effluent by using a second solvent, and filtering the redissolved effluent to obtain filtrate; and measuring the filtrate by using ultra performance liquid chromatography-tandem mass spectrometry. The method can simply, quickly, efficiently, accurately and environmentally detect the residual quantity of the imazethapyr in the milk or bean beverage.
Description
Technical Field
The invention belongs to the technical field of food detection, and particularly relates to a method for detecting imazethapyr residue in milk or bean beverage.
Background
Imazapic is a high-efficiency and high-selectivity herbicide, and the wide application of imazapic causes the problems of biological toxicity, environmental pollution, pesticide residue and the like, thereby bringing about potential safety hazards of food. The pesticide can accumulate in animal and human body and cause acute or chronic poisoning through the enrichment of food chain, and seriously threatens human health. With increasing importance on food safety, the maximum limit of residual imazapic in food is definitely set to be 0.01-0.3mg/kg by the national release of the maximum limit of residual pesticide in national food Standard for food safety (GB 2763) -2019 in 8 and 15 days, wherein the maximum residual imazapic in raw milk is 0.01mg/kg, and the maximum residual imazapic in beans is 0.3 mg/kg.
However, the analysis samples suitable for the existing imazapic detection method mainly comprise soil, water, medicinal plants, fruits, vegetables, grains and the like, and the detection reports of the milk and the bean beverage are few, so that the establishment of the method for detecting the residual imazapic in the milk and the bean beverage has important significance.
At present, the detection method of the imazethapyr mainly comprises a gas chromatography-mass spectrometry combined method and a liquid chromatography, and the pretreatment mainly comprises a derivation method, a dispersion solid phase extraction method and a gel chromatography. The derivation method has complex operation steps, strict experimental condition requirements and more impurities on the machine; the dispersive solid-phase extraction method is simple and quick in technology, but cannot process complex matrix samples; gel chromatography can process complex matrices, but the operation is time consuming and consumes more solvent. Therefore, the development of a simple, fast, efficient, accurate and environment-friendly technology suitable for detecting the residual quantity of imazethapyr in milk and bean drinks is urgently needed.
Disclosure of Invention
The invention aims to provide a method for detecting residual quantity of imazapic in milk or bean drinks, which promotes protein precipitation in the milk or bean drinks by adopting an acidified organic solvent, and removes impurities better through solid phase extraction and purification, so that the purity of the extracted imazapic is higher, and the detection result is quicker and more accurate.
According to one aspect of the invention, the method for detecting the residual quantity of imazamox in the milk or bean beverage comprises the following steps:
mixing a sample to be detected with an acidified first solvent to obtain a mixture so as to extract residual imazapic in the sample to be detected;
adding sodium chloride into the mixture, uniformly mixing, centrifuging, and collecting supernatant;
carrying out solid phase extraction on the supernatant, and collecting all effluent liquid;
drying the effluent, redissolving the effluent by using a second solvent, and filtering the redissolved effluent to obtain filtrate;
and measuring the filtrate by using ultra performance liquid chromatography-tandem mass spectrometry.
Preferably, the first solvent is acetonitrile, and the acid used to acidify the first solvent is one or both of formic acid and acetic acid; the second solvent is acetonitrile-water mixed solution.
Preferably, the volume ratio of the acid to the first solvent is (0.001-0.01):1, and the volume ratio of acetonitrile to water in the second solvent is (0.2-1.0): 1.
Preferably, the mass ratio of the acidified first solvent to the sample to be detected is (2.5-10):1, and the amount of the sodium chloride is 0.1-1.0 time of the mass of the sample to be detected.
Preferably, the solid phase extraction step is carried out using a solid phase extraction column packed with lipophilic divinylbenzene and hydrophilic N-vinylpyrrolidone, and using the acidified first solvent as eluent.
Preferably, the solid phase extraction column is pre-activated with the acidified first solvent.
Preferably, the specification of the solid phase extraction column is 60-300 mg.
Preferably, the centrifugation step is carried out at a low temperature of 0-8 ℃.
Preferably, the milk is raw or processed milk secreted by a mammal, and the soy drink is a soy drink obtained by processing beans.
Preferably, the conditions of the measurement parameters of the ultra performance liquid chromatography-tandem mass spectrometry are as follows:
ultra-high performance liquid chromatography conditions:
column ACQUITY UPLC BEH C181.7 μm, 2.1X 50mm, or a column of comparable performance; column temperature: 35 ℃; sample introduction amount: 3 mu L of the solution; the mobile phase A is 0.1% formic acid aqueous solution or 5mmol ammonium formate aqueous solution, the mobile phase B is acetonitrile, the flow rate is 0.3 mL/min; gradient elution procedure: 95% A at 0-1min, 95-20% A at 1-3min, 20% A at 3-4min, 5% A at 4-5min, and 95% A at 5-8 min;
tandem mass spectrometry conditions:
ionization mode: electrospray negative ion mode (ESI +); capillary voltage: 1.13 kV; removing the solvent gas: nitrogen gas; collision gas: argon gas; desolventizing agent gas flow: 800L/Hr; taper hole gas flow: 50L/Hr; source temperature: 120 ℃; desolventizing temperature: 350 ℃; taper hole voltage: 38V.
The determination method disclosed by the invention has the advantages of simple pretreatment operation, short time consumption, less used organic solvent, good extraction and separation effects, quick and accurate detection result and very strong application value.
Drawings
Further objects, features and advantages of the present invention will become apparent from the following description of embodiments of the invention, with reference to the accompanying drawings, in which:
FIG. 1 shows a chromatogram of imazethapyr residue in milk measured by ultra high performance liquid chromatography-tandem mass spectrometry in accordance with example 1, at an added concentration of 5. mu.g/kg.
Detailed Description
Technical features, objects and advantages of the present invention will be more clearly understood and appreciated by those skilled in the art. It should be understood that the following detailed description is merely exemplary, and the technical solution of the present invention is not limited to the specific embodiments listed below.
The method for detecting imidazole nicotinic acid residue in milk or bean beverage provided by the invention comprises an extraction step, a centrifugal separation step, a purification step and a determination step, wherein the purification step comprises a solid phase extraction step and a redissolution step.
The milk detected by the invention refers to unprocessed or processed milk secreted by mammals, and comprises but is not limited to raw milk, raw goat milk, pasteurized milk, ultrahigh-temperature sterilized milk, modified milk and the like; the detected bean beverage is a bean beverage obtained by processing beans, and includes but is not limited to a bean beverage obtained by processing and treating red beans, mung beans, soybeans, black beans or green beans serving as raw materials.
Wherein the extraction step comprises mixing a sample to be tested with an acidified first solvent to obtain a mixture, so as to extract residual imazapyr in the sample to be tested.
The first solvent is preferably acetonitrile. The acid used to acidify the first solvent may be one or both of formic acid and acetic acid, with formic acid being preferred. Wherein the volume ratio of the acid to the first solvent is (0.001-0.01):1, preferably (0.003-0.007):1, more preferably (0.004-0.006): 1. Further, it is preferable to use formylated acetonitrile in the extraction step.
The centrifugation step comprises adding sodium chloride to the mixture obtained in the extraction step, preferably by vortexing, and centrifuging to collect the supernatant. Centrifugation is preferably carried out at 15000 r/min. The centrifugation step is carried out at cryogenic conditions, preferably at 0-8 ℃.
According to a specific embodiment, the extraction step may be: weighing 2.00g of a sample to be detected in a 30mL centrifuge tube, adding 10mL of acidified acetonitrile mixed solution, fully performing vortex oscillation, adding 0.50g of sodium chloride, fully performing vortex oscillation, and centrifuging at the temperature of 4 ℃ at 15000r/min for 10min to obtain a supernatant.
The inventor of the invention determines the extraction method by screening to ensure that the residual quantity of imazethapyr in milk and bean drinks can be sensitively, accurately and effectively detected.
According to GB/T23818-. According to GB/T20769 + 2008 < determination of residual quantity of 450 pesticides and related chemicals in fruits and vegetables >, acetonitrile is adopted for extraction, the extraction efficiency of the imazapic in milk is 15% -25%, and the extraction efficiency is too low.
The invention adopts acidified acetonitrile mixed solution for extraction, and is based on the following reasons: the imazapyr is an amphoteric compound, mainly exists in a molecular state under an acidic condition, and is beneficial to extraction by an organic solvent; secondly, the milk and the bean beverage matrix contain certain fat and large protein molecules, the acetonitrile can greatly reduce the extraction of fat and some lipophilic pigments in the sample matrix, and has good precipitation effect on the large protein molecules, thereby being beneficial to the subsequent purification.
The mixed solution of formic acid and acetonitrile is used as an extraction reagent, the content level of imidazole nicotinic acid which can be detected is 0.005mg/kg-0.03mg/kg, and the extraction recovery rate is more than 80%.
Different proportions of the acidified acetonitrile mixed liquor can influence the extraction effect on the imazethapyr, the acidity is too low, the extraction rate on the imazethapyr is low, the acidity is too high, and the extraction rate is not obviously improved. When the volume ratio of the acid to the acetonitrile is (0.001-0.01):1, the extraction efficiency is more than 70%, and when the volume ratio of the acid to the acetonitrile is (0.004-0.006):1, the extraction efficiency is more than 80%, and the volume ratio of the acid to the acetonitrile is 0.004-0.006, so that the extraction rate can be ensured, and the use amount of the acid can be saved.
The solid phase extraction step comprises passing the supernatant through a solid phase extraction column. Preferably, the supernatant is transferred to an activated solid phase extraction column and eluted with an acidified first solvent, and the entire effluent is collected. The solid phase extraction column can be an HLB solid phase extraction column, wherein the filler can be lipophilic divinylbenzene and hydrophilic N-vinyl pyrrolidone, and the specification is preferably 60-300mg, preferably 100-280mg, more preferably 150-250mg, and most preferably 200 mg. The solid phase extraction column is activated by wetting with an acidified first solvent.
According to a specific embodiment, raw milk is used as a matrix, the acidified acetonitrile mixed solution is used as an extracting solution, the extracting solution is purified by an HLB (hydrophile-lipophile balance) solid-phase extraction column, and the acidified acetonitrile is used for elution, so that the recovery rate of the imazethapyr can reach 70-90%. Under the same conditions, if a conventional amino solid phase extraction column (specification is 1g) is adopted for purification, the recovery rate is less than 30 percent by respectively eluting with acidified acetonitrile, dichloromethane and acetonitrile.
The re-dissolving step comprises blow-drying the effluent, preferably with nitrogen, to remove the acidified first solvent from the effluent, specifically, the blow-drying temperature of nitrogen may be 40 deg.C; and re-dissolving the effluent with a second solvent after drying. The second solvent may be an acetonitrile-water mixture, preferably the volume ratio of acetonitrile to water is (0.2-1.0):1, preferably (0.4-0.8):1, for example 0.6: 1.
According to another embodiment, the purification step may be: accurately measuring 5mL of supernatant obtained in the extraction step, transferring the supernatant to a solid phase extraction column activated by 3mL of formic acid-acetonitrile mixed solution, eluting the solid phase extraction column by using 3mL of formic acid-acetonitrile mixed solution, combining effluent, blowing nitrogen at 40 ℃ to be nearly dry, redissolving by using acetonitrile-water mixed solution to 1mL, carrying out vortex oscillation for 1min, transferring the supernatant to a 1.5mL centrifuge tube, centrifuging at 4 ℃ for 5min at 20000r/min, taking the supernatant through a 0.22 mu m filter membrane, and taking filtrate for ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).
The determination step is carried out by using an ultra-high performance liquid chromatography-tandem mass spectrometer. The parameter conditions of the ultra performance liquid chromatography-tandem mass spectrometry are as follows.
Ultra-high performance liquid chromatography conditions:
the chromatographic column adopts ACQUITY UPLC BEH C181.7 μm of Waters company, 2.1 × 50mm, or equivalent performance chromatographic column; column temperature: 35 ℃; sample introduction amount: 3 mu L of the solution; the mobile phase A is 0.1% formic acid aqueous solution or 5mmol ammonium formate aqueous solution, the mobile phase B is acetonitrile, the flow rate is 0.3 mL/min; gradient elution procedure: 95% A at 0-1min, 95-20% A at 1-3min, 20% A at 3-4min, 5% A at 4-5min, and 95% A at 5-8 min;
tandem mass spectrometry conditions:
ionization mode: electrospray negative ion mode (ESI +); capillary voltage: 1.13 kV; removing the solvent gas: nitrogen gas; collision gas: high-purity argon gas; desolventizing agent gas flow: 800L/Hr; taper hole gas flow: 50L/Hr; source temperature: 120 ℃; desolventizing temperature: 350 ℃; taper hole voltage: 38V.
In conclusion, the method for measuring the imazethapyr residual quantity is simple and feasible, can be used for detecting the imazethapyr residual quantity in the milk and bean beverage, or can be used as a means for controlling the imazethapyr residual quantity in the milk and bean beverage to be in a safe range, and has the advantages of rapidness, accuracy and certain popularization value.
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments.
The methods used in the following examples are conventional methods unless otherwise specified, and the reagents used are commercially available reagents unless otherwise specified.
Examples
Example 1 detection of imazethapyr residue in milk
Selecting raw milk produced in a certain pasture, respectively adding 3 standard substance solutions of 5 microgram/kg, 10 microgram/kg and 30 microgram/kg into the raw milk, extracting, purifying and detecting. The specific method comprises the following steps:
1) extraction: weighing 2.00g of raw milk sample into a 30mL centrifuge tube, adding imazapic standard solution, shaking up, adding 10mL formic acid-acetonitrile mixed solution (the volume ratio of formic acid to acetonitrile is 0.005:1), fully vortexing and shaking, adding 0.50g of sodium chloride, fully vortexing and shaking, and centrifuging at the temperature of 4 ℃ at 15000r/min for 10min to obtain supernatant.
2) Purifying: accurately measuring 5mL of the supernatant obtained in the step 1), transferring the supernatant to an HLB solid-phase extraction column activated by 3mL of formic acid-acetonitrile mixed solution, controlling the flow rate at 1mL/min, eluting the solid-phase extraction column by using 3mL of formic acid-acetonitrile mixed solution (the volume ratio of formic acid to acetonitrile is 0.005:1), collecting all eluent, drying the eluent by using nitrogen at 40 ℃, redissolving the eluent to 1mL by using acetonitrile-water mixed solution (the volume ratio of acetonitrile to water is 0.25:1), carrying out vortex oscillation for 1min, transferring the eluent to a 1.5mL centrifuge tube, centrifuging for 5min at 4 ℃ 20000r/min, taking the supernatant, and filtering the supernatant by using a 0.22 mu m filter membrane to obtain a filtrate.
Wherein the HLB solid phase extraction column is filled with lipophilic divinylbenzene and hydrophilic N-vinyl pyrrolidone filler, and the specification is 200 mg.
3) Detecting the filtrate by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)
Liquid chromatography conditions: a chromatographic column: ACQUITY UPLC BEH C181.7 μm, 2.1X 50mm (Waters Corp.); column temperature: 35 ℃; sample introduction amount: 3 mu L of the solution; the mobile phase A is 0.1% formic acid water solution, the mobile phase B is acetonitrile, and the flow rate is 0.3 mL/min; gradient elution procedure: 95% A at 0-1min, 95-20% A at 1-3min, 20% A at 3-4min, 5% A at 4-5min, and 95% A at 5-8 min.
Mass spectrum conditions: ionization mode: electrospray negative ion mode (ESI +); capillary voltage: 1.13 kV; removing the solvent gas: nitrogen gas; collision gas: high-purity argon gas; desolventizing agent gas flow: 800L/Hr; taper hole gas flow: 50L/Hr; source temperature: 120 ℃; desolventizing temperature: 350 ℃; taper hole voltage: 38V.
According to the embodiment, 3 horizontal imazapyr standard solutions are added to a sample to be detected, and the recovery rate and RSD value of the imazapyr are obtained through ultra performance liquid chromatography-tandem mass spectrometry, as shown in Table 1.
Table 1 mean recovery and relative standard deviation of raw milk n ═ 6
According to this example, the imazapyr multireaction monitoring (MRM) chromatogram obtained by adding 5 μ g/kg of standard solution to raw milk as determined by ultra performance liquid chromatography-tandem mass spectrometry is shown in fig. 1, wherein 262.04>217.07 is the quantitative ion chromatogram (upper); 262.04>234.18 is a qualitative ion chromatogram (below).
Example 2 detection of Imidazonicotinic acid residue in Bean-based beverage
Selecting some commercial prepared soybean milk (original taste), adding 3 standard solutions of 5. mu.g/kg, 10. mu.g/kg and 30. mu.g/kg respectively, extracting, purifying, and detecting. The specific method comprises the following steps:
1) extraction: weighing 2.00g of prepared soybean milk (original flavor) sample in a 30mL centrifuge tube, adding imidazole nicotinic acid standard solution, shaking up, adding 10mL formic acid-acetonitrile mixed solution (the volume ratio of formic acid to acetonitrile is 0.005:1), fully vortexing and shaking, adding 0.50g of sodium chloride, fully vortexing and shaking, and centrifuging at 4 ℃ for 10min at 15000r/min to obtain a supernatant.
2) Purifying: accurately measuring 5mL of the supernatant obtained in the step 1), transferring the supernatant to an HLB solid-phase extraction column activated by 3mL of formic acid-acetonitrile mixed solution, controlling the flow rate at 1mL/min, eluting the solid-phase extraction column by using 3mL of formic acid-acetonitrile mixed solution (the volume ratio of formic acid to acetonitrile is 0.005:1), collecting all eluent, drying the eluent at 40 ℃ by using nitrogen, redissolving the acetonitrile-water mixed solution (the volume ratio of acetonitrile to water is 0.25:1) to 1mL, carrying out vortex oscillation for 1min, transferring the eluent to a 1.5mL centrifuge tube, centrifuging for 5min at 4 ℃ at 20000r/min, taking the supernatant, and filtering the supernatant through a 0.22 mu m filter membrane to obtain a filtrate.
Wherein the HLB solid-phase extraction column is filled with lipophilic divinylbenzene and hydrophilic N-vinyl pyrrolidone filler, and the specification is 200 mg.
3) Detecting the filtrate by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)
Liquid phase conditions: a chromatographic column: ACQUITY UPLC BEH C181.7 μm, 2.1X 50mm (Waters Corp.), or equivalent; column temperature: 35 ℃; sample introduction amount: 3 mu L of the solution; the mobile phase A is 0.1% formic acid water solution, the mobile phase B is acetonitrile, and the flow rate is 0.3 mL/min; gradient elution procedure: 95% A at 0-1min, 95-20% A at 1-3min, 20% A at 3-4min, 5% A at 4-5min, and 95% A at 5-8 min.
Mass spectrum conditions: ionization mode: electrospray negative ion mode (ESI +); capillary voltage: 1.13 kV; removing the solvent gas: nitrogen gas; collision gas: high-purity argon gas; desolventizing agent gas flow: 800L/Hr; taper hole gas flow: 50L/Hr; source temperature: 120 ℃; desolventizing temperature: 350 ℃; taper hole voltage: 38V.
According to the embodiment, 3 horizontal imazapic standard solutions are added into a sample to be detected, and the recovery rate and RSD value of the imazapic are obtained through ultra performance liquid chromatography tandem mass spectrometry measurement and are shown in table 2.
Table 2 mean recovery and relative standard deviation n ═ 6 for conditioned soy milk (original flavor) base
Comparative example 1 detection of imazethapyr residue in milk
1) Extraction: weighing 2.00g of raw milk sample produced in a certain pasture into a 30mL centrifuge tube, adding imazapic standard solution, shaking up, adding 10mL formic acid-acetonitrile mixed solution (the volume ratio of formic acid to acetonitrile is 0.005:1), fully vortexing and shaking, adding 0.50g of sodium chloride, fully vortexing and shaking, and centrifuging at 4 ℃ for 10min at 15000r/min to obtain supernatant.
2) Purifying: accurately measuring 5mL of the supernatant obtained in the step 1), transferring the supernatant to an amino solid-phase extraction column activated by 3mL of formic acid-acetonitrile mixed solution, controlling the flow rate at 1mL/min, eluting the solid-phase extraction column by using 3mL of formic acid-acetonitrile mixed solution (the volume ratio of formic acid to acetonitrile is 0.005:1), collecting all eluent, drying the eluent by using nitrogen at 40 ℃, redissolving the eluent to 1mL by using acetonitrile-water mixed solution (the volume ratio of acetonitrile to water is 0.25:1), carrying out vortex oscillation for 1min, transferring the eluent to a 1.5mL centrifuge tube, centrifuging for 5min at 4 ℃ 20000r/min, taking the supernatant, and filtering the supernatant by using a 0.22 mu m filter membrane to obtain a filtrate.
Wherein, the specification of the amino solid phase extraction column is 1 g.
3) Detecting the filtrate by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)
Liquid chromatography conditions: a chromatographic column: ACQUITY UPLC BEH C181.7 μm, 2.1 × 50 mm; column temperature: 35 ℃; sample introduction amount: 3 mu L of the solution; the mobile phase A is 0.1% formic acid water solution, the mobile phase B is acetonitrile, and the flow rate is 0.3 mL/min; gradient elution procedure: 95% A at 0-1min, 95-20% A at 1-3min, 20% A at 3-4min, 5% A at 4-5min, and 95% A at 5-8 min.
Mass spectrum conditions: ionization mode: electrospray negative ion mode (ESI +); capillary voltage: 1.13 kV; removing the solvent gas: nitrogen gas; collision gas: high-purity argon gas; desolventizing agent gas flow: 800L/Hr; taper hole gas flow: 50L/Hr; source temperature: 120 ℃; desolventizing temperature: 350 ℃; taper hole voltage: 38V.
According to the comparative example, the imazapic standard solutions with the levels of 5 mug/kg (n 6) and 10 mug/kg (n 6) are added into the sample to be tested, and the recovery rate of the imazapic is lower than 30% through the ultra performance liquid chromatography-tandem mass spectrometry.
The foregoing is only a preferred embodiment of the present invention. It will be appreciated that various modifications, combinations, alterations, and substitutions of the details and features of the invention may be made by those skilled in the art without departing from the spirit and nature of the invention. Such modifications, combinations, alterations and substitutions are also to be understood as being included within the scope of the invention as claimed.
Claims (10)
1. A method for detecting residual quantity of imazethapyr in milk or bean beverage is characterized by comprising the following steps:
mixing a sample to be detected with an acidified first solvent to obtain a mixture so as to extract residual imazapic in the sample to be detected;
adding sodium chloride into the mixture, uniformly mixing, centrifuging, and collecting supernatant;
carrying out solid phase extraction on the supernatant, and collecting all effluent liquid;
drying the effluent, redissolving the effluent by using a second solvent, and filtering the redissolved effluent to obtain filtrate;
and measuring the filtrate by using ultra performance liquid chromatography-tandem mass spectrometry.
2. The detection method according to claim 1, wherein the first solvent is acetonitrile, and the acid for acidifying the first solvent is one or both of formic acid and acetic acid; the second solvent is acetonitrile-water mixed solution.
3. The detection method according to claim 2, wherein the volume ratio of the acid to the first solvent is (0.001-0.01):1, and the volume ratio of acetonitrile to water in the second solvent is (0.2-1.0): 1.
4. The detection method according to claim 1, wherein the volume ratio of the acidified first solvent to the sample to be detected is (2.5-10):1, and the amount of the sodium chloride is 0.1-1.0 times of the mass of the sample to be detected.
5. The detection method according to claim 1, wherein the solid phase extraction step is performed using a solid phase extraction column packed with lipophilic divinylbenzene and hydrophilic N-vinylpyrrolidone, and the acidified first solvent is used as an eluent.
6. The detection method of claim 5, wherein the solid phase extraction column is pre-activated with the acidified first solvent.
7. The detection method according to claim 5 or 6, wherein the specification of the solid phase extraction column is 60-300 mg.
8. The detection method according to claim 1, wherein the centrifugation step is performed at a low temperature of 0 to 8 ℃.
9. The detection method according to claim 1, wherein the milk is unprocessed or processed milk secreted from a mammal, and the soy drink is a soy drink obtained by processing beans.
10. The detection method according to claim 1, wherein the parameter conditions of the ultra performance liquid chromatography-tandem mass spectrometry are as follows:
ultra-high performance liquid chromatography conditions:
column ACQUITY UPLC BEH C181.7 μm, 2.1X 50mm, or a column of comparable performance; column temperature: 35 ℃; sample introduction amount: 3 mu L of the solution; the mobile phase A is 0.1% formic acid aqueous solution or 5mmol ammonium formate aqueous solution, the mobile phase B is acetonitrile, the flow rate is 0.3 mL/min; gradient elution procedure: 95% A at 0-1min, 95-20% A at 1-3min, 20% A at 3-4min, 5% A at 4-5min, and 95% A at 5-8 min;
tandem mass spectrometry conditions:
ionization mode: electrospray negative ion mode ESI +; capillary voltage: 1.13 kV; removing the solvent gas: nitrogen gas; collision gas: argon gas; desolventizing agent gas flow: 800L/Hr; taper hole gas flow: 50L/Hr; source temperature: 120 ℃; desolventizing temperature: 350 ℃; taper hole voltage: 38V.
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| CN111157651A (en) * | 2020-01-08 | 2020-05-15 | 石家庄君乐宝乳业有限公司 | Method for detecting content of residual pesticide in dairy product |
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| CN111157651A (en) * | 2020-01-08 | 2020-05-15 | 石家庄君乐宝乳业有限公司 | Method for detecting content of residual pesticide in dairy product |
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