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CN-122017081-A - Method for detecting 12 hppd inhibitor herbicides in plant-derived food

CN122017081ACN 122017081 ACN122017081 ACN 122017081ACN-122017081-A

Abstract

The invention belongs to the technical field of HPPD inhibitor herbicide residue detection in plant source foods, and particularly relates to a detection method for 12 HPPD inhibitor herbicides in plant source foods, which comprises the steps of 1, weighing solid standard products of 12 target compounds, preparing single standard stock solution, diluting with acetonitrile-water to prepare mixed standard intermediate solution, 2, preparing a plant source food sample to be detected, 3, weighing the sample in the 2, adding acetonitrile formate, sodium chloride, swirling and centrifuging, absorbing supernatant into a centrifuge tube containing anhydrous magnesium sulfate and purifying materials, 4, performing LC-MS/MS detection, namely, adopting positive ions to scan simultaneously, separating by using a C18 chromatographic column, taking the formic acid-acetonitrile as a mobile phase, and performing multiple reaction monitoring mode (MRM) detection.

Inventors

  • SU YOUZHI
  • LEI HONGQIN
  • LI FANG
  • LI YANMEI
  • LIU JUN

Assignees

  • 伊宁海关技术中心

Dates

Publication Date
20260512
Application Date
20260323

Claims (8)

  1. 1. The method for detecting 12 hppd inhibitor herbicides in plant-derived food is characterized by comprising the following steps: respectively and accurately weighing solid standard substances of 12 target compounds, namely Mesotrione (MES), cyclosulfamone (TEM), topramezone (BBC), sulcotrione (SUL), pyraflufen-ethyl (PSF), clomazone (PZL), isoxaflutole (ISO), amicarbazone (TEF), dicyclopyrone (BIC), pyrazolote (PZF), a liquid standard stock solution Topramezone (TOP) and pyraclon (BFP), dissolving the solid standard stock solution with methanol, preparing a single standard stock solution, diluting the single standard stock solution and the liquid standard stock solution with acetonitrile-water, and preparing a mixed standard intermediate solution; Step 2, preparing a plant source food sample to be detected, namely homogenizing a fruit and vegetable sample masher, randomly sampling, crushing grains, sieving, preserving the prepared sample at-18 ℃ and below, and placing the sample at room temperature until the sample is completely thawed before detection; step 3, weighing the sample in the step 2, adding acetonitrile formate, swirling, adding sodium chloride, swirling and centrifuging, sucking supernatant into a centrifuge tube containing anhydrous magnesium sulfate and a purifying material, swirling and centrifuging, sucking supernatant liquid, blowing the supernatant to dryness, fixing the volume by acetonitrile-water (1:9 v/v), swirling and coating to be detected, wherein the purifying material is a water-wettable polymer (40-60 mu m) with a PAX strong anion exchange mixing mechanism, and purifying by two adsorbents (40-60 mu m) together; and 4, LC-MS/MS detection, namely separating by adopting a positive ion simultaneous scanning mode and a C18 chromatographic column, taking formic acid-acetonitrile as a mobile phase, and detecting by adopting a multiple reaction monitoring mode (MRM), wherein the substrate is quantitatively matched with an external standard method.
  2. 2. The method for detecting the herbicide of 12 types hppd inhibitors in the plant-derived food, which is characterized in that in the step 1, the solid standard substances 10 mg (accurate to 0.01 and mg) of 12 types of target compounds are respectively and accurately weighed in a brown volumetric flask of 10mL, dissolved by methanol and fixed to a scale, a single standard stock solution with the mass concentration of 1000 mug/mL is prepared, the single standard stock solution and the liquid standard stock solution are diluted by acetonitrile-water (1:9 v/v) to prepare a mixed standard intermediate solution with the mass concentration of 1.0 mug/mL, and the mixed standard intermediate solution is stored at the temperature of-20 ℃.
  3. 3. The method for detecting the herbicide of 12 types of hppd inhibitors in plant-derived foods according to the claim 2, wherein in the step 2, 9 substrate samples of millet, corn, soybean, rice, flaxseed, wheat, potato, tomato, grape and the like are adopted, the substrate analysis solution is obtained after extraction and purification, the substrate analysis solution is used for gradually diluting and mixing the standard intermediate solution until the concentration of each compound is 10 ng/mL, the standard substrate analysis solution is obtained, after the 9 blank samples are extracted and purified, 1mL of 0.1, 0.2, 0.5, 1, 5, 10, 20, 50, 100 ng/mL standard solution is respectively used for volume fixation, the substrate matching standard curve is obtained, and the linear regression equation is carried out on the mass concentration (x) of the pesticide according to the peak area (y).
  4. 4. The method for detecting 12 kinds of hppd inhibitor herbicides in a plant-derived food according to claim 1, wherein in the step 3, the volume ratio of the amounts of the two adsorbents of C18 and PAX is C18 120~150mg,PAX 120~150 mg.
  5. 5. The method for detecting 12 kinds of hppd inhibitor herbicides in a plant-derived food according to claim 3, wherein in step 3, a sample 5g (grain and oil crop samples 2.5g+7.5ml of water, potato samples 5g+3ml of water) with a relatively large water content is weighed into a 50mL plastic centrifuge tube, 10 mL of 1% acetonitrile formate is added, vortexing is performed for 1min, and human 3g sodium chloride is added, vortexing is performed for 1min,8000 r/min, and centrifugation is performed for 1 min. Absorbing supernatant 3mL to a 15 mL plastic centrifuge tube (PAX 150mg, C18 mg, 200mg anhydrous magnesium sulfate) containing a water removing agent and a purifying material, swirling 30s, centrifuging 3 min at 10000 r/min, absorbing supernatant 2 mL, blowing nitrogen to dryness in a 40 oC water bath, fixing volume by using 1 mL acetonitrile-water (1:9 v/v), swirling 60s, passing through a 0.22 mu m nylon filter membrane, and measuring filtrate.
  6. 6. The method for detecting 12 kinds of hppd inhibitor herbicides in a plant-derived food according to claim 1, wherein in the step 4, LC-MS/MS conditions are InfinityLab poroshell 120, 120 EC-C18 column (3.1. 3.1 mm.times.100 mm,2.7 μm), column temperature of 35 ℃, sample introduction amount of 10.0. Mu.L, mobile phase A of 0.1% formic acid solution (containing 5mmol/L ammonium acetate), B of acetonitrile, and flow rate of 0.3 mL/min.
  7. 7. The method for detecting the hppd inhibitor herbicides in the plant-derived food according to claim 6, wherein in the step 4, the elution procedure is as follows, 0 to 1.0 min (10% of phase B), 1.0 to 2.0 min (10% to 50% of phase B), 2.0 to 10.0 min (50% to 95% of phase B), 11.0 to 11.1 min (95% to 10% of phase B), and 11.1 to 15 min (10% of phase B).
  8. 8. The method for detecting 12 kinds of hppd inhibitor herbicides in a plant-derived food according to claim 6, wherein in the step 4, the ionization mode is electrospray positive ion mode ionization (ESI+), multiple Reaction Monitoring (MRM), drying gas temperature is 350 ℃, drying gas flow rate is 10L/min, sheath gas temperature is 300 ℃, sheath gas flow rate is 9L/min, and capillary voltage is 4000V.

Description

Method for detecting 12 hppd inhibitor herbicides in plant-derived food Technical Field The invention belongs to the technical field of hppd inhibitor herbicide residue detection in plant-derived foods, and particularly relates to a detection method of 12 hppd inhibitor herbicides in plant-derived foods. Background The HPPD inhibitor herbicide is fully called a p-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor, and can block the conversion of p-hydroxyphenylpyruvate into homogentisate by inhibiting the activity of HPPD, so that tocopherol, plastoquinone, carotenoid and the like cannot be normally synthesized, plant meristem, new tissues are promoted to generate albino symptoms, and further, the plants are dead (Fu, qi, ng et al Jugulam), the HPPD inhibitor herbicide has the advantages of high activity, wide weed control spectrum, good safety and the like, is widely used for preventing and treating crop weeds, can be divided into 6 types of trione compounds (topramezone, dicyclopyrone, mesotrione, sulcotrione, biscarfentrazone and cyclosulfamone), 5 types of pyrazolones (pyriftone, benflumetofen, pyrazolone, topramezone, isoxazophos) and 1 type of isoxazolone (isoxazophos), wherein the market of the HPPD inhibitor herbicide has relatively high activity, can be used as a medicament for treating the acute tyrosin, parkinsonism or other potential neurodegenerative diseases, has the toxic and harmful effects on the skin and the skin of rats, and the growth of the rats, and the toxic and the harmful effects on the growth of the water-borne algae are promoted. The analysis method of the residual quantity of the HPPD inhibitor herbicide mainly comprises a fluorescent probe analysis method, a spectrophotometry method, an electrochemical method, a liquid chromatography method and a liquid chromatography-time-of-flight mass spectrometry, wherein in the prior art, the fluorescent probe analysis method, the spectrophotometry method, the electrochemical method and the liquid chromatography method can not carry out confirmatory analysis on the compound, the qualitative aspect has a defect, the residual quantity of the HPPD inhibitor herbicide is freshly reported by adopting a gas chromatograph-mass spectrometry combined technology, the liquid chromatography-time-of-flight mass spectrometry is mainly used for quick screening of the compound, the quantitative aspect has a defect, the liquid chromatography-tandem mass spectrometry can accurately quantify the compound, can also carry out confirmatory qualitative analysis on the compound, and the detection method in the prior art can only carry out detection on the residual quantity of the individual HPPD inhibitor herbicides such as topramezone, isoxazodone and topramezone in plant source foods such as corn, wheat and rice, and lacks a method capable of simultaneously meeting the requirement of detecting the 12 HPPD inhibitor herbicides, so the method for carrying out the maximum supervision on the residual quantity of the HPPD inhibitor herbicide in foods is allowed to carry out the maximum limit quantity monitoring on the residual quantity of the HPPD inhibitor herbicide (MRPD). Disclosure of Invention Aiming at the problems, the invention aims to provide an improved QuEChERS pretreatment technology, and an ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry combined technology is combined to establish an analysis method of the residual quantity of 12 HPPD inhibitor herbicides in plant-derived foods, which is simple, convenient, quick, economical, low in detection limit, good in accuracy and precision, and provides technical support for simultaneously monitoring the residual quantity of 12 HPPD inhibitor herbicides. The invention aims at realizing the technical scheme that the method for detecting the hppd inhibitor herbicides in the plant source food comprises the following steps: respectively and accurately weighing solid standard substances of 12 target compounds, namely Mesotrione (MES), cyclosulfamone (TEM), topramezone (BBC), sulcotrione (SUL), pyraflufen-ethyl (PSF), clomazone (PZL), isoxaflutole (ISO), amicarbazone (TEF), dicyclopyrone (BIC), pyrazolote (PZF), a liquid standard stock solution Topramezone (TOP) and pyraclon (BFP), dissolving the solid standard stock solution with methanol, preparing a single standard stock solution, diluting the single standard stock solution and the liquid standard stock solution with acetonitrile-water, and preparing a mixed standard intermediate solution; Step 2, preparing plant source food samples to be detected, namely homogenizing the fruit and vegetable samples by a masher, placing the homogenized fruit and vegetable samples in a polyethylene bottle, randomly sampling, crushing grains, sieving the crushed grains, placing the crushed grains in the polyethylene bottle or bag, preserving the prepared samples at the temperature of-18 ℃ or below, placing all frozen samples in a room temperature condi