Method for quantitatively determining the residues of buprofezin and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid-phase extraction

Abstract

The present invention discloses a method for quantitatively determining the residues of bipropionate and its metabolite M440I007 in broken tea infusion by PVPP combined with Cleanert TPT solid-phase extraction, which includes steps such as sample extraction, sample purification, UPLC-MS / MS determination, blank test, result calculation and expression, and judgment. First of all, the present invention purifies the test sample through the solid-phase dispersion of PVPP and the Cleanert TPT solid-phase extraction column, which can effectively purify the co-extracted substances that interfere with the detection of the target substance in broken tea infusion; secondly, on this basis, the present invention adopts specific UPLC-MS / MS detection conditions for the target substance; finally, it not only has high extraction efficiency and excellent purification effect, but also can meet the detection requirements in terms of recovery rate, detection limit and precision.

Description

Technical Field

[0001] The present invention belongs to the technical field of detection, and specifically relates to a method for determining the residues of bipropyl dicarboxylate and its metabolite M440I007 in broken tea infusion by PVPP combined with Cleanert TPT solid-phase extraction method. Background Art

[0002] Bipropyl dicarboxylate is a new type of biogenic insecticide, which was registered for the control of Empoasca vitis on tea trees in September 2023. It will metabolize on tea leaves to produce M440I007 with similar toxicity. At present, there is little research on the detection methods of bipropyl dicarboxylate and M440I007 in tea. The existing literature reports only focus on the detection in fresh tea leaves, dry tea, tea tree roots, stems and whole tea infusions, and this method cannot meet the requirements of the recovery rate in broken tea infusion. Due to the lack of analysis methods for bipropyl dicarboxylate and M440I007 in broken tea infusion, it is impossible to monitor the residue content, leaching rate and dietary risk assessment in broken tea infusion. Therefore, it is of great significance to establish an analysis method for bipropyl dicarboxylate and M440I007 in broken tea infusion.

[0003] The molecular structural formulas of bipropyl dicarboxylate (A) and metabolite M440I007 (B) are shown as follows:

[0004] Summary of the Invention

[0005] Aiming at the problems existing in the prior art, the purpose of the present invention is to design and provide a technical scheme for quantitatively determining the residues of bipropyl dicarboxylate and its metabolite M440I007 in broken tea infusion by PVPP (crosslinked polyvinylpyrrolidone) combined with Cleanert TPT solid-phase extraction method. This method can effectively purify the co-extracted substances that interfere with the detection of the target substances in broken tea infusion through the solid-phase dispersion of PVPP and the purification of Cleanert TPT solid-phase extraction column. This method is easy to operate, has excellent purification effect, and can meet the requirements of residue detection in terms of recovery rate, detection limit and precision, and can provide technical support for the determination of bipropyl dicarboxylate and M440I007 in broken tea infusion.

[0006] In order to achieve the above purpose, the present invention specifically adopts the following technical solutions:

[0007] A method for quantitatively determining the residues of bipropyl dicarboxylate and its metabolite M440I007 in broken tea infusion by PVPP combined with Cleanert TPT solid-phase extraction method, comprising the following steps:

[0008] 1) Sample extraction

[0009] Preparation of green tea, black tea and oolong tea infusions: Crush green tea, black tea and oolong tea, brew with water, and filter to obtain the infusions; After extraction with acetonitrile, add PVPP (cross-linked polyvinylpyrrolidone) for purification, centrifuge and take the supernatant, repeat the extraction once, combine the supernatants and rotary evaporate to dryness; Then re-dissolve with a mixed solvent of acetonitrile and benzene, filter through a membrane, and wait for purification;

[0010] 2) Sample purification

[0011] Take a Cleanert TPT column, pre-rinse with a mixed solvent of acetonitrile and benzene, load the re-dissolved solution obtained in step 1) onto the column, elute with a mixed solvent of acetonitrile and benzene, collect all the loaded solution and eluate, rotary evaporate to dryness, and make up the volume with acetonitrile for UPLC-MS / MS testing;

[0012] 3) UPLC-MS / MS determination

[0013] Inject the matrix-matched standard solution and the sample solution under the following set liquid chromatography - mass spectrometry / mass spectrometry conditions:

[0014] A) High performance liquid chromatography - mass spectrometry / mass spectrometry instrument parameters:

[0015] High performance liquid chromatography: Use an Acquity UPLC HSS-T3 chromatographic column for chromatographic separation;

[0016] Mass spectrometry conditions: Analyze the bipropylpylate and M440I007 using the multiple reaction monitoring mode MRM and the ESI + mode;

[0017] B) Mass spectrometry parameters of bipropylpylate and metabolite M440I007

[0018]

[0019] In the table represents the quantitative ion;

[0020] 4) Blank test

[0021] Except for not adding the sample, operate according to the above steps 1), 2), and 3);

[0022] 5) Result calculation and expression

[0023] Calculate the mass concentration of bipropylpylate and metabolite M440I007 in the sample;

[0024] 6) Judgment

[0025] Plot the standard working curve. If the retention time of the chromatographic peak of the analyte in the sample solution is consistent with that of the matrix-matched standard solution, it can be determined that the target substance is contained.

[0026] Further, the specific steps for sample extraction in step 1) are as follows:

[0027] Crush green tea, black tea, and oolong tea, brew them at a tea-to-water ratio of 1:50, and filter to obtain the tea soup. Take 20 mL of the tea soup into a 50 mL polytetrafluoroethylene centrifuge tube, add 20 mL of acetonitrile, mix well, add 200 mg of PVPP, vortex for 5 min, add 7 g of NaCl, vortex for 2 min, centrifuge at 5000 r / min for 5 min, and take all the supernatant. Repeat the extraction once, combine the supernatants, and rotary evaporate to dryness. Re-dissolve with 5 mL of a mixed solvent of acetonitrile:benzene with a volume ratio of 3:1, filter through a 0.22 μm filter membrane, and wait for purification.

[0028] Further, the specific steps for sample purification in step 2) are as follows:

[0029] Pre-rinse the Cleanert TPT column with 5 mL of a mixed solvent of acetonitrile:benzene with a volume ratio of 3:1, load the re-dissolved solution onto the column, elute with 30 mL of a mixed solvent of acetonitrile:benzene with a volume ratio of 3:1, collect all the loaded solution and the eluate, rotary evaporate to dryness at 40°C, and dissolve and make up the volume to 1 mL with acetonitrile for UPLC-MS / MS testing.

[0030] Further, in step 3), the chromatographic column specifications are 100 mm × 2.1 µm, 1.8 µm; Waters, Milford, MA, USA.

[0031] Further, in step 3), the high-performance liquid chromatography conditions are as follows: the mobile phase is 0.1% formic acid in acetonitrile (A) and water (B), the flow rate is 0.25 mL / min, and the gradient elution program is 60% - 99% A from 0.0 - 3.0 min, 99% A from 3.0 - 4.5 min, 60% A from 5.0 - 6.5 min; the column temperature is 40°C, and the injection volume is 5 μL.

[0032] Further, in step 3), the mass spectrometry conditions are as follows: electrospray voltage: 3.0 kV; ion source temperature: 150°C; desolvation temperature: 350°C; the desolvation gas and the cone hole counter gas are nitrogen, and the flow rates are 700 L / hr and 60 L / hr respectively; the collision gas (Ar) flow rate is 0.35 mL / min.

[0033] Furthermore, in step 5), the formula for calculating the mass concentrations of the buprofezin and metabolite M440I007 in the sample is as follows, and the blank value must be deducted from the processing result:

[0034]

[0035] In the formula:

[0036] Xi ── Residual amount of the target substance in the sample, unit: milligram per kilogram;

[0037] C i ── Concentration of the target substance solution obtained from the standard curve, unit: milligram per liter;

[0038] V ── Final volume of the sample solution after constant volume, unit: milliliter;

[0039] m ── Mass of the test sample represented by the final sample solution, unit: gram.

[0040] Furthermore, step 6) specifically includes:

[0041] Taking the mass concentration X as the abscissa and the peak area ratio Y as the ordinate, draw a 5-point standard working curve, and use the standard working curve to quantitatively determine the sample. The response value of the analyte in the sample solution meets the linear range of the instrument detection; both the quantitative and qualitative ion pairs are present, and the signal-to-noise ratio ≥ 3; the retention time of the chromatographic peak of the analyte in the sample solution is consistent with that of the matrix-matched standard solution, and the allowable deviation is less than ±0.5%, then it can be determined that the target substance is contained.

[0042] The determination method of the present invention has the following beneficial effects:

[0043] The present invention first purifies the test sample through the solid-phase dispersion of PVPP and the Cleanert TPT solid-phase extraction column, which can effectively purify the co-extracted substances in the broken tea tea soup that interfere with the detection of the target substance; secondly, on this basis, the present invention adopts specific UPLC-MS / MS detection conditions for the target substance; finally, not only the extraction efficiency is high and the purification effect is excellent, but also the recovery rate, detection limit and precision can meet the detection requirements. Description of the Drawings

[0044] Figure 1 Mass spectra of the standards of bipropylproxyphene and M440I007, concentration 0.0008 mg / L;

[0045] Figure 2 Effect of the amount of PVPP on the recovery rate of bipropylproxyphene and M440I007. Specific Embodiments

[0046] The following further illustrates the present invention with reference to the embodiments.

[0047] Example 1: Establishment of mass spectrometry conditions

[0048] ESI +In the [mode], the full-scan method was used to obtain the parent ions of bipropyl dicylopropionate and M440I007, which were m / z 594.2 and 1187.8 respectively. The cone voltage (20 - 80 V) was further optimized. The responses of bipropyl dicylopropionate and M440I007 were the highest at 62 V and 35 V respectively. The daughter ion scan of the parent ions was performed by tandem mass spectrometry, and the collision dissociation energy (10 - 70 eV) was optimized. The characteristic daughter ions with the highest ion abundances of bipropyl dicylopropionate were m / z 148.0 and 68.9, and the optimal collision energies were 52 V and 50 V respectively. There was no matrix interference at m / z 594.2>148.0 and 594.2>68.9. Therefore, the ion pair with the highest response, m / z 594.2>148.0, was selected as the quantitative ion pair, and 594.2>68.9 was selected as the qualitative ion pair. The qualitative and quantitative ion pairs of metabolite M440I007 were optimized in the same way. The mass spectrometry diagram is shown in the appendix Figure 1 .

[0049] Example 2: Detection of bipropyl dicylopropionate and M44I007 in broken tea infusion by Cleanert TPT solid-phase extraction method

[0050] The recoveries of bipropyl dicylopropionate and M44I007 in broken tea infusion were directly detected by Cleanert TPT solid-phase extraction method, and the results are shown in Table 1. When only the Cleanert TPT column was used to purify the broken tea infusion, the recoveries of bipropyl dicylopropionate in the broken tea infusions of 3 tea types were lower than 77.18%, and the recovery in the broken tea matrix of oolong tea was the lowest, only 36.92%. The recoveries of M440I007 in the broken tea infusions of 3 tea types were lower than 78.12%, and the recovery in the broken tea matrix of black tea was the lowest, only 62.49%. It could not meet the recovery requirements in pesticide residue analysis, and the pretreatment method needed to be optimized.

[0051] Table 1 shows the recoveries of bipropyl dicylopropionate and M44I007 in broken tea infusion detected by Cleanert TPT solid-phase extraction method (spiking level is 0.2 or 2 mg / kg, n = 3)

[0052]

[0053] Example 3: Detection of bipropyl dicylopropionate and M44I007 in broken tea infusion by PVPP dispersive solid-phase extraction method

[0054] Taking the broken oolong tea soup as an example, the recoveries of bipropionate and M44I007 in the broken tea soup were compared between the method of only using PVPP for dispersive solid-phase extraction and the method of combining PVPP with Cleanert TPT solid-phase extraction. The results are shown in Table 2. When only PVPP dispersive solid-phase extraction was used for purification, the recoveries of bipropionate and M440I007 were 67.04% and 65.53% respectively; when PVPP was combined with Cleanert TPT solid-phase extraction, the recoveries of the two target substances in the broken tea matrix were both increased to more than 80%. Therefore, PVPP was used in succession with Cleanert TPT solid-phase extraction for the pretreatment of the broken tea matrix.

[0055] Table 2 shows the recoveries of bipropionate and M44I007 in the broken tea soup detected by PVPP dispersive solid-phase extraction and PVPP combined with Cleanert TPT solid-phase extraction (the addition level is 0.2 mg / kg, n = 3)

[0056]

[0057] Example 4: Optimization of PVPP dosage

[0058] Taking the broken oolong tea soup as an example, the initial purification of the broken tea soup was carried out with the PVPP dosage set at 0, 50, 100, 200, 300 and 400 mg, and then combined with Cleanert TPT solid-phase extraction. It was found that as the PVPP dosage increased from 0 - 50 mg to 200 mg, the recovery rate increased significantly (P < 0.05). When the PVPP dosage was 200 mg, the recovery rate of bipropionate reached the highest at 88.15% (RSD = 5.89%); there was no significant difference in the recovery rate of M440I007 when the PVPP dosage was 200 - 400 mg (P > 0.05). Therefore, 200 mg PVPP was selected as the initial purification filler in the broken tea soup matrix.

[0059] 1. Reagents and materials

[0060] 1.1 Acetonitrile: Chromatographic grade.

[0061] 1.2 Benzene: Chromatographic grade.

[0062] 1.3 Anhydrous sodium chloride: Analytical grade.

[0063] 1.4 PVPP (CAS: 25294 - 54 - 1) USP grade.

[0064] 1.5 Cleanert TPT solid-phase extraction column: (1 g / 6 mL).

[0065] 1.6 Standard substance of afidopyropen (Afidopyropen, CAS: 915972–17-7, C 33 H 39 NO 9 ): 98.7%.

[0066] 1.7 Standard substance of M440I007 (C 65 H 76 N 2 O 18 ): Purity 90.2%.

[0067] 1.8 Standard stock solution: Accurately weigh 0.010 g (accurate to 0.0001 g) of each of the standard substances (1.6 and 1.7) into a 50 mL volumetric flask, dilute to the mark with acetonitrile to prepare a 100 mg / L stock standard solution, and store it at -20 °C.

[0068] 1.9 Mixed standard solution: Accurately measure 5 mL (accurate to 0.01 mL) of the standard stock solution (1.8) into a 25 mL volumetric flask, dilute to the mark with acetonitrile to prepare a 20 mg / L stock standard solution, and store it at -20 °C.

[0069] 1.10 Preparation of standard intermediate solution: Accurately pipette an appropriate amount of the standard stock solution (1.9), and use acetonitrile to prepare a 4 mg / L mixed standard solution, which is stored in the dark in a -20 °C refrigerator.

[0070] 1.11 Preparation of standard working solution: Dilute the standard intermediate solution (1.10) with acetonitrile to an appropriate concentration of the standard working solution as needed.

[0071] 1.12 Microporous filter membrane: 0.22 μm, organic phase.

[0072] 2. Instruments and equipment

[0073] 2.1 High performance liquid chromatography - mass spectrometry / mass spectrometry instrument: Equipped with (ESI) ion source.

[0074] 2.2 Analytical balance: Sensitivity 0.00001 g and 0.01 g.

[0075] 2.3 Vortex mixer.

[0076] 2.4 Solid sample grinder.

[0077] 2.5 Centrifuge.

[0078] 2.6 Ultrasonic instrument.

[0079] 3. Determination steps

[0080] 3.1 Sample extraction

[0081] Preparation of green tea, black tea and oolong tea broths: The green tea, black tea and oolong tea were crushed and brewed at a tea-to-water ratio of 1:50, and the broth was obtained by filtration. Take 20 mL of the broth into a 50 mL polytetrafluoroethylene centrifuge tube, add 20 mL of acetonitrile, mix well, add 200 mg of PVPP, vortex for 5 min, add 7 g of NaCl, vortex for 2 min, centrifuge at 5000 r / min for 5 min, and take all the supernatant. Repeat the extraction once, combine the supernatants and rotary evaporate to dryness. Re-dissolve with 5 mL of acetonitrile:benzene 3:1 (v:v), filter through a 0.22 μm filter membrane, and wait for purification.

[0082] 3.2 Sample purification

[0083] The Cleanert TPT column was pre-eluted with 5 mL of acetonitrile:benzene 3:1 (v:v), the re-dissolved solution was loaded onto the column, eluted with 30 mL of acetonitrile:benzene 3:1 (v:v), all the loaded solution and eluate were collected, rotary evaporated to dryness at 40 °C, and made up to volume with 1 mL of acetonitrile for UPLC-MS / MS testing.

[0084] 3.3 Determination

[0085] Since the test conditions depend on the instrument used, it is impossible to give general parameters for chromatographic analysis. The following operating conditions have been proven to be suitable for the test. Specific parameters are shown in Tables 3 and 4 in the attached table.

[0086] Table 3 Liquid chromatography parameters

[0087]

[0088] Table 4 Mass spectrometry condition parameters

[0089]

[0090] 4. Blank test

[0091] All steps were carried out as described above except that no sample was added.

[0092] 5. Result calculation and expression

[0093] Calculate the contents of bipropylphenylpyrazole and M440I007 in the broken tea broth samples according to the following formula, and the processing results must be deducted from the blank value:

[0094]

[0095] In the formula:

[0096] X i ── The residue amount of the target substance in the sample, in milligrams per kilogram;

[0097] C i ── The solution concentration of the analyte obtained from the standard curve, in mg / L;

[0098] V ── The final volume of the sample solution after dilution, in mL;

[0099] m ── The mass of the sample represented by the final sample solution, in g.

[0100] Determine the contents of bipropyl dicylopropionate and M440I007 in broken tea tea soup matrix by UPLC-MS / MS. For preparing the standard curve, measure the peak areas of the standard samples at 0.0008 mg / L, 0.002 mg / L, 0.01 mg / L, 0.04 mg / L, 0.4 mg / L, 1 mg / L, and 2 mg / L respectively. Plot the standard curves with the response values against the contents of bipropyl dicylopropionate and M440I007 as y = 671719x - 3764.9 (R 2 = 0.9999) and y = 1,251,345x - 14,599 (R 2 = 1). The peak area value of the sample to be tested is 1000. Substitute it into the standard curve for calculation. C 双丙环虫酯 and C M440I007 are 0.007 mg / L and 0.012 mg / L respectively. V is 1 mL and m is 0.4 g. Substitute into the formula for calculation. X 双丙环虫酯 = 0.018 mg / kg, X M440I007 = 0.031 mg / kg.

[0101] 6. Limit of determination

[0102] The limit of determination of this method is 0.002 mg / kg.

[0103] 7. Recovery rate

[0104] Three spiked concentrations were set for the recovery experiment of this method. According to the experimental conditions determined by this method, conduct 5 experiments for each spiked concentration. The results of the recovery rate and precision are shown in Table 5.

[0105] Table 5 Spiked concentrations and recovery rate ranges of bipropyl dicylopropionate and M440I007 in broken tea tea soup

[0106]

[0107] Example 5: Application example

[0108] Collect samples 5 days after applying bipropyl dicylopropionate (5% bipropyl dicylopropionate dispersible concentrate, application rate 22.5 g a.i. / hm 2The fresh tea leaves of () are made into oolong tea. According to the method of the present invention, the residues of bipropylpicarzin and its metabolites in the oolong tea broken tea soup are determined, and the leaching rate is calculated. The leaching rate of bipropylpicarzin is 16.0% - 20.7%; the leaching rate of metabolite M440I007 is 4.3% - 6.0%.

[0109] The above are the preferred embodiments of the present invention, which are only used to explain the present invention and are not used to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. PVPP combined with Cleanert TPT solid phase extraction method for quantitative determination of the residues of diprofenopterin and its metabolite M440I007 in broken tea soup. The molecular structure of the metabolite M440I007 is shown below: ， It is characterized in that The following steps are involved: 1) Sample extraction Preparation of green tea, black tea and oolong tea soup: crush green tea, black tea and oolong tea, brew with water, filter to obtain tea soup; extract with acetonitrile, add PVPP for purification, take the supernatant after centrifugation, repeat the extraction once, combine the supernatant and evaporate to dryness; then re-dissolve with a mixed solvent of acetonitrile and benzene, filter through a membrane, and wait for purification; 2) Sample purification Take a Cleanert TPT column, pre-elute it with a mixed solvent of acetonitrile and benzene, load the complex solution obtained in step 1) onto the column, elute it with a mixed solvent of acetonitrile and benzene, collect all the column liquid and the eluate, rotary evaporate to dryness, and make up the volume with acetonitrile for UPLC-MS / MS testing; 3) UPLC-MS / MS determination The matrix-matched standard solution and sample solution were injected under the following LC-MS / MS conditions: A) HPLC-MS / MS instrument parameters: High performance liquid chromatography: Acquity UPLC HSS-T3 column was used for chromatographic separation; Mass spectrometry conditions: Multiple reaction monitoring mode MRM and ESI + The pattern was used to analyze abicyclon and M440I007; B) Mass spectrometric parameters of abicyclon and its metabolite M440I007 Table represents the quantitative ion; 4) Blank test Except that no sample is added, the above steps 1), 2) and 3) are followed; 5) Calculation and presentation of results Calculate the mass concentrations of abicyclon and metabolite M440I007 in the samples; 6) Judgment A standard working curve was drawn. If the retention time of the chromatographic peak of the analyte in the sample solution was consistent with that of the matrix-matched standard solution, it could be determined that the sample contained acyclovir and its metabolite M440I007.

2. The method for quantitatively determining the residues of diprofenoprene and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid phase extraction as claimed in claim 1, characterized in that: The step 1) sample extraction specifically includes: Green tea, black tea and oolong tea were crushed, brewed at a tea-water ratio of 1:50, and filtered to obtain tea soup; 20 mL of tea soup was taken into a 50 mL polytetrafluoroethylene centrifuge tube, 20 mL of acetonitrile was added, mixed, 200 mg of PVPP was added, vortexed for 5 min, 7 g of NaCl was added, vortexed for 2 min, centrifuged at 5000 r / min for 5 min, and all the supernatant was taken; the extraction was repeated once, the supernatants were combined and rotary evaporated to dryness; re-dissolved with 5 mL of a mixed solvent with a volume ratio of acetonitrile:benzene of 3:1, filtered through a 0.22 μm filter membrane, and purified.

3. The method for quantitatively determining the residues of diprofenoprene and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid phase extraction as claimed in claim 1, characterized in that: The step 2) sample purification specifically includes: The Cleanert TPT column was pre-eluted with 5 mL of a mixed solvent of acetonitrile:benzene in a volume ratio of 3:

1. The complex solution was loaded onto the column and eluted with 30 mL of a mixed solvent of acetonitrile:benzene in a volume ratio of 3:

1. All the column liquid and the eluent were collected and rotary evaporated to dryness at 40°C. The volume was fixed with 1 mL of acetonitrile for UPLC-MS / MS testing.

4. The method for quantitatively determining the residues of diprofenoprene and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid phase extraction as claimed in claim 1, characterized in that: The specifications of the chromatographic column in step 3) are 100 mm×2.1 µm, 1.8 µm; Waters, Milford, MA, USA.

5. The method for quantitatively determining the residues of diprofenoprene and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid phase extraction as claimed in claim 1, characterized in that: In the step 3), the high performance liquid chromatography is as follows: the mobile phase is 0.1% formic acid acetonitrile A and water B, the flow rate is 0.25 mL / min, the gradient elution program is 60%-99% A 0.0-3.0 min, 99% A 3.0-4.5 min, 60% A 5.0-6.5 min; the column temperature is 40°C, and the injection volume is 5 μL.

6. The method for quantitatively determining the residues of diprofenoprene and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid phase extraction as claimed in claim 1, characterized in that: The mass spectrometry conditions in step 3) are as follows: electrospray voltage: 3.0 kV; ion source temperature: 150°C; desolvation temperature: 350°C; desolvation gas and cone backflush gas are nitrogen, with flow rates of 700 L / hr and 60 L / hr, respectively; Collision gas Ar flow rate: 0.35 mL / min.

7. The method for quantitatively determining the residues of diprofenoprene and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid phase extraction as claimed in claim 1, characterized in that: The calculation and expression of the results in step 5) are as follows: The formula for calculating the mass concentration of abicyclon and metabolite M440I007 in the sample is as follows, and the blank value must be deducted from the processing result: Where: X i ——The residual amount of the target substance in the sample, in milligrams per kilogram; C i ——The concentration of the target solution obtained from the standard curve, in milligrams per liter; V – the final volume of the sample solution, in milliliters; m – the mass of the sample represented by the final sample solution, in grams.

8. The method for quantitatively determining the residues of diprofenoprene and its metabolite M440I007 in broken tea soup by PVPP combined with Cleanert TPT solid phase extraction as claimed in claim 1, characterized in that: The judgment in step 6) specifically includes: using mass concentration X as the horizontal axis and peak area ratio Y as the vertical axis, a 5-point standard working curve is drawn, and the sample is quantitatively determined using the standard working curve. The response value of the analyte in the sample solution meets the linear range of instrument detection; quantitative and qualitative ion pairs exist at the same time, and the signal-to-noise ratio is ≥3; the retention time of the chromatographic peak of the analyte in the sample solution is consistent with that of the matrix matching standard solution, and the allowable deviation is less than ±0.5%, then it can be determined that the target substance is contained.

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