Method for analyzing and detecting residual quantity of novel neonicotinoid insecticide in plasma, tissue organs, urine and excrement of mammal

Abstract

The invention discloses a method for analyzing and detecting the residual quantity of novel neonicotinoid insecticides in plasma, tissue organs, urine and faeces of mammals, and belongs to the technical field of pesticide detection.The method comprises the following steps that S1, a plasma / tissue organ / urine / faeces sample of the mammals is pretreated; and S2, carrying out on-machine detection on the treated sample. Through a series of experimental operation steps of organic solvent extraction, solid-phase extraction, organic solvent elution and nitrogen blowing concentration, the invention provides a pretreatment method which is simple to operate, high in recovery rate, good in stability and suitable for treating plasma, tissue organs, urine and excrement samples of mammals. And then carrying out quantitative analysis on the target insecticide by using a sample obtained by pretreatment by using liquid chromatography Ultimate 3000 connected in series with orbitrap high-resolution mass spectrometry Q-Exactive. The analysis and detection method provided by the invention provides reference for evaluating the internal exposure level of the novel neonicotinoid insecticide in mammals.

Description

Technical Field

[0001] The present invention belongs to the technical field of pesticide detection, and in particular relates to an analytical detection method for the residues of novel neonicotinoid insecticides in mammalian plasma, tissues and organs, urine and feces, and adopts liquid chromatography Ultimate 3000 tandem orbital trap high-resolution mass spectrometry Q-Exactive to determine the residues of novel neonicotinoid insecticides in various matrix samples. Background Art

[0002] Neonicotinoids are a class of insecticides following organochlorine insecticides, organophosphorus insecticides, carbamate insecticides and pyrethroid insecticides. They can selectively bind to the nicotinic acetylcholine receptors in insects, affect the insect's central nervous system, and cause the insect to be continuously excited and paralyzed, thereby achieving the effect of killing insects. However, due to the widespread use of traditional neonicotinoid insecticides, a large number of target insects have developed different levels of resistance to them, and caused the death of some beneficial insects such as bees or a sharp decline in the number of insectivorous birds. Therefore, new neonicotinoid insecticides came into being.

[0003] New neonicotinoid insecticides have higher insecticidal activity against insects that are resistant to neonicotinoid insecticides, and have lower toxicity to insects such as bees. Among them, two new neonicotinoid insecticides, chlorothiazide and flonicamid, were registered in my country in 2002 and 2007, respectively, accounting for about 5% and 10% of the Chinese market share, respectively. Due to their wide application and low volatility and high water solubility, these insecticides are constantly detected in different environmental media around the world.

[0004] Currently, research on new neonicotinoid insecticides is mostly focused on their residue levels in the environment and crops and their toxicity and safety evaluation on insects, while research on their internal exposure levels in mammals is still scarce, and relevant analytical detection methods urgently need to be established. Summary of the invention

[0005] The purpose of the present invention is to provide an analytical detection method for the residues of new neonicotinoid insecticides in mammalian plasma, tissues and organs, urine and feces, which has simple operation, good stability and high recovery rate, and can be applied to the study of the internal exposure level of new neonicotinoid insecticides in mammals.

[0006] In order to achieve the above purpose, this application adopts the following technical solutions:

[0007] The present invention provides a method for analyzing and detecting the residue of a novel neonicotinoid insecticide in mammalian plasma, tissue organs, urine, and feces, comprising the following steps:

[0008] S1: Pretreatment of mammalian plasma / tissue / urine / feces samples;

[0009] Among them, the pretreatment method of mammalian plasma is:

[0010] After thawing the mammalian plasma sample, transfer the plasma sample to a centrifuge tube, add acetonitrile to precipitate protein, vortex and shake, and then centrifuge to obtain a supernatant;

[0011] The supernatant is concentrated and purified by using an HLB SPE solid phase extraction column. First, the HLB SPE solid phase extraction column is activated with Milli-Q water and an organic solvent. Then, the supernatant is passed through the HLB SPE solid phase extraction column and then the HLB SPE solid phase extraction column is eluted with an organic solvent, and the eluate is collected.

[0012] The eluate is concentrated by nitrogen blowing under a nitrogen flow, and then the volume is fixed with a solvent. The resulting solution is then centrifuged and the supernatant is removed for testing on an instrument.

[0013] The pre-treatment method of mammalian tissues and organs is:

[0014] Weigh the homogenized tissue and organ samples and add an organic solvent, shake and centrifuge to obtain a supernatant;

[0015] The supernatant was concentrated and purified using an HLB SPE solid phase extraction column. First, the HLB SPE solid phase extraction column was activated with Milli-Q water and an organic solvent, and then the supernatant was passed through the HLB SPE solid phase extraction column and the effluent was collected;

[0016] The collected liquid was concentrated by nitrogen blowing under a nitrogen flow, and then the volume was fixed with a solvent. The resulting solution was then centrifuged and the supernatant was removed for testing on an instrument.

[0017] The pretreatment method of mammalian urine is:

[0018] After the mammalian urine sample is thawed, the urine sample is transferred to an HLB SPE solid phase extraction column for concentration and purification. First, the HLB SPE solid phase extraction column is activated with an organic solvent and Milli-Q water. Then, the urine sample is passed through the HLB SPE solid phase extraction column and then the HLB SPE solid phase extraction column is eluted with an organic solvent, and the eluate is collected.

[0019] The eluate is then blown to near dryness under a nitrogen stream, and then the volume is fixed with a solvent. The resulting solution is then centrifuged, and the supernatant is removed for testing on an instrument.

[0020] The pre-treatment methods for mammal feces are:

[0021] After freeze-drying, the feces of mammals are fully ground, the feces sample is weighed and an organic solvent is added, and after shaking and centrifugation, a supernatant is obtained;

[0022] The supernatant was concentrated and purified using an HLB SPE solid phase extraction column. First, the HLB SPE solid phase extraction column was activated with Milli-Q water and an organic solvent, and then the supernatant was passed through the HLB SPE solid phase extraction column and the effluent was collected;

[0023] The collected liquid was concentrated by nitrogen blowing under a nitrogen flow, and then the volume was fixed with a solvent. The resulting solution was then centrifuged and the supernatant was removed for testing on an instrument.

[0024] S2: The processed samples are tested on the machine. The specific instrument analysis method is as follows:

[0025] The target compounds were quantitatively analyzed using liquid chromatography Ultimate 3000 tandem orbitrap high-resolution mass spectrometry Q-Exactive.

[0026] In the above technical solution, in S2, the chromatographic column used is Hypersil Gold TM C18 column, pre-column is Hypersil Gold TM C18, the mobile phases were Milli-Q water containing 0.1% formic acid and acetonitrile, respectively. The mass spectrometer obtained full scan mass spectral data in positive ionization mode with a scan range of 100-750Da and a resolution of 140,000. High-purity nitrogen was used as sheath gas and auxiliary gas. The sheath gas pressure and auxiliary gas pressure were 35psi and 15arb, respectively, and the spray voltage was +3000V. The ion transfer tube temperature was 350°C, the H-ESI heating temperature was 300°C, and an electrospray ion source was used.

[0027] In the above technical solution, the mammal is one of a mouse, a cat, a dog, a cow, a sheep, a rabbit, a donkey and a horse.

[0028] In the above technical solution, the time for freeze-drying mammal feces is 5-7 days.

[0029] In the above technical scheme, the organic solvent used in the solid phase extraction process is one of acetonitrile, acetonitrile containing 0.1% formic acid, and acetonitrile: dichloromethane with a volume ratio of 3:7. Different organic solvents are selected according to different pesticides.

[0030] In the above technical solution, the solvent used for constant volume is acetonitrile.

[0031] The beneficial effect of the present invention is that: the present invention provides a pretreatment method suitable for processing mammalian plasma, tissues and organs, urine and feces samples with simple operation, high recovery rate and good stability through a series of experimental operation steps of organic solvent extraction, solid phase extraction, organic solvent elution and nitrogen blowing concentration, and then the samples obtained by pretreatment are quantitatively analyzed for target insecticides using liquid chromatography Ultimate 3000 tandem orbital trap high-resolution mass spectrometry Q-Exactive. The analytical detection method of the present invention provides a reference for evaluating the internal exposure level of new neonicotinoid insecticides in mammals. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is a flow chart of the method for pre-treating mammalian plasma / tissue organ / urine / feces samples according to the present invention;

[0033] Figure 2 It is the standard curve diagram of chlorothiophene of the present invention;

[0034] Figure 3 The chromatogram and mass spectrum obtained in Example 1 of the present invention are shown. DETAILED DESCRIPTION

[0035] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. The present invention can be implemented in many different forms and should not be construed as being limited to the embodiments set forth herein. On the contrary, these embodiments are provided so that this disclosure will be thorough and complete, and the concept of the present invention will be fully conveyed to those skilled in the art, and the present invention will only be limited by the claims.

[0036] Example 1

[0037] The present embodiment relates to a standard curve of chlorothiophene, specifically comprising the following steps:

[0038] The purchased chlorothiophene standard was taken and diluted with chromatography grade acetonitrile to prepare a mixed standard series of 5, 10, 20, 50 and 100 μg / L respectively.

[0039] The target compound chlorothioline was quantitatively analyzed by liquid chromatography Ultimate 3000 tandem orbitrap high-resolution mass spectrometry Q-Exactive (Liquid Chromatogram-Quadrupole-Orbitrap High-Resolution Mass Spectrometer, ThermoFisher Scientific, USA). The chromatographic column used was Hypersil Gold TMC18 chromatographic column (100 mm × 2.1 mm × 3 μm; Thermo Scientific, USA), pre-column: Hypersil Gold TM C18 (10mm×2.1mm×3μm). The mobile phases were Milli-Q water (A) and acetonitrile (B) containing 0.1% formic acid, respectively. The mobile phase gradient program was 10% B from 0 to 1min, changed to 100% B within 5min and maintained for 4min, and changed to 10% B within 2min and maintained for 4min. The injection volume was 5μL and the column temperature was 40°C. The mass spectrometer obtained full scan mass spectral data in positive ionization mode with a scan range of 100-750Da and a resolution of 140,000. After each full scan, the top 10 ions with the strongest response values ​​were fully scanned for secondary daughter ions. The resolution of the secondary scan was 17,500, the collision energy used was 30%, 40% and 50%, and the dynamic exclusion time was set to 3 seconds. High-purity nitrogen was used as sheath gas and auxiliary gas. The sheath gas pressure and auxiliary gas pressure were 35psi and 15arb, respectively. The spray voltage was +3000V. The ion transfer tube temperature was 350°C, and the H-ESI heating temperature was 300°C. An electrospray ion source was used.

[0040] Example 2

[0041] This embodiment relates to a method for analyzing and detecting the residual amount of chlorthiazide in rat plasma, which specifically comprises the following steps:

[0042] S1 Pre-treatment of plasma samples

[0043] (1) 50 μL of plasma was removed and 200 μL of acetonitrile was added, vortexed for 1 min, shaken on an oscillator at 250 rpm for 30 min, and then centrifuged at 1300 g in a high-speed centrifuge for 10 min to obtain the supernatant.

[0044] (2) First, activate the HLB SPE (60 mg / 3 mL) column with 3 mL of Milli-Q water and 3 mL of acetonitrile, then pass the supernatant obtained in step (1) through the column, and then elute the SPE column with 5 mL of acetonitrile, and collect the eluate.

[0045] (3) The eluate obtained in step (2) was nitrogen-blown to 0.5 mL under a gentle nitrogen flow (purity: 99%). The resulting solution was then centrifuged at 9500 g for 5 min, and the supernatant was removed for testing.

[0046] S2 Instrumental Analysis Method

[0047] The target compound chlorothioline was quantitatively analyzed using a liquid chromatography Ultimate 3000 tandem orbitrap high-resolution mass spectrometer Q-Exactive (Liquid Chromatogram-Quadrupole-Orbitrap High-Resolution Mass Spectrometer, ThermoFisher Scientific, USA). The chromatographic column used was Hypersil Gold TM C18 chromatographic column (100 mm × 2.1 mm × 3 μm; Thermo Scientific, USA), pre-column: Hypersil Gold TM C18 (10mm×2.1mm×3μm). The mobile phases were Milli-Q water (A) and acetonitrile (B) containing 0.1% formic acid, respectively. The mobile phase gradient program was 10% B from 0 to 1 min, changed to 100% B within 5 min and maintained for 4 min, and changed to 10% B within 2 min and maintained for 4 min. The injection volume was 5μL and the column temperature was 40°C. The mass spectrometer obtained full scan mass spectral data in positive ionization mode with a scan range of 100-750Da and a resolution of 140,000. After each full scan, the top 10 ions with the strongest response values ​​were fully scanned for secondary daughter ions. The resolution of the secondary scan was 17,500, the collision energy used was 30%, 40% and 50%, and the dynamic exclusion time was set to 3 seconds. High-purity nitrogen was used as sheath gas and auxiliary gas. The sheath gas pressure and auxiliary gas pressure were 35psi and 15arb, respectively. The spray voltage was +3000V. The ion transfer tube temperature was 350°C, and the H-ESI heating temperature was 300°C. An electrospray ion source was used.

[0048] Example 3

[0049] This embodiment relates to a method for analyzing and detecting the residual amount of chlorthiazide in rat tissues and organs, which specifically comprises the following steps:

[0050] S1 Pre-processing of tissue and organ samples

[0051] (1) After the tissues and organs are homogenized, 0.5 g is weighed and 2 mL of acetonitrile is added. The mixture is shaken at 250 rpm for 30 min in an oscillator, and then centrifuged at 1300 g for 10 min in a high-speed centrifuge to obtain a supernatant. The above steps are repeated three times and the supernatants are combined.

[0052] (2) First, activate the HLB SPE (60 mg / 3 mL) column with 3 mL of Milli-Q water and 3 mL of acetonitrile, then pass the supernatant obtained in step (1) through the column and collect the effluent.

[0053] (3) The solution obtained in step (2) was nitrogen-blown to 0.5 mL under a gentle nitrogen flow (purity: 99%). The obtained solution was then centrifuged at 9500 g for 5 min, and the supernatant was removed for testing.

[0054] S2 Instrumental Analysis Method

[0055] The target compound chlorothioline was quantitatively analyzed using a liquid chromatography Ultimate 3000 tandem orbitrap high-resolution mass spectrometer Q-Exactive (Liquid Chromatogram-Quadrupole-Orbitrap High-Resolution Mass Spectrometer, ThermoFisher Scientific, USA). The chromatographic column used was Hypersil Gold TM C18 chromatographic column (100 mm × 2.1 mm × 3 μm; Thermo Scientific, USA), pre-column: Hypersil Gold TM C18 (10mm×2.1mm×3μm). The mobile phases were Milli-Q water (A) and acetonitrile (B) containing 0.1% formic acid, respectively. The mobile phase gradient program was 10% B from 0 to 1min, changed to 100% B within 5min and maintained for 4min, and changed to 10% B within 2min and maintained for 4min. The injection volume was 5μL and the column temperature was 40°C. The mass spectrometer obtained full scan mass spectral data in positive ionization mode with a scan range of 100-750Da and a resolution of 140,000. After each full scan, the top 10 ions with the strongest response values ​​were fully scanned for secondary daughter ions. The resolution of the secondary scan was 17,500, the collision energy used was 30%, 40% and 50%, and the dynamic exclusion time was set to 3 seconds. High-purity nitrogen was used as sheath gas and auxiliary gas. The sheath gas pressure and auxiliary gas pressure were 35psi and 15arb, respectively. The spray voltage was +3000V. The ion transfer tube temperature was 350°C, and the H-ESI heating temperature was 300°C. An electrospray ion source was used.

[0056] Example 4

[0057] This embodiment relates to a method for analyzing and detecting the residual amount of chlorthiazide in rat urine, which specifically comprises the following steps:

[0058] S1 Pre-treatment of urine samples

[0059] (1) Pipette 0.5 mL of the supernatant and concentrate and purify it using an HLB SPE solid phase extraction cartridge.

[0060] (2) First, activate the HLB SPE (60 mg / 3 mL) column with 3 mL of acetonitrile and 3 mL of Milli-Q water, then pass the urine obtained in step (1) through the column, and then elute with 5 mL of acetonitrile, and collect the eluate.

[0061] (3) The eluate obtained in step (2) was nitrogen-blown to 0.5 mL under a gentle nitrogen flow (purity: 99%). The resulting solution was then centrifuged at 9500 g for 5 min, and the supernatant was removed for testing.

[0062] S2 Instrumental Analysis Method

[0063] The target compound chlorothioline was quantitatively analyzed using a liquid chromatography Ultimate 3000 tandem orbitrap high-resolution mass spectrometer Q-Exactive (Liquid Chromatogram-Quadrupole-Orbitrap High-Resolution Mass Spectrometer, ThermoFisher Scientific, USA). The chromatographic column used was Hypersil Gold TM C18 chromatographic column (100 mm × 2.1 mm × 3 μm; Thermo Scientific, USA), pre-column: Hypersil Gold TM C18 (10mm×2.1mm×3μm). The mobile phases were Milli-Q water (A) and acetonitrile (B) containing 0.1% formic acid, respectively. The mobile phase gradient program was 10% B from 0 to 1 min, changed to 100% B within 5 min and maintained for 4 min, and changed to 10% B within 2 min and maintained for 4 min. The injection volume was 5μL and the column temperature was 40°C. The mass spectrometer obtained full scan mass spectral data in positive ionization mode with a scan range of 100-750Da and a resolution of 140,000. After each full scan, the top 10 ions with the strongest response values ​​were fully scanned for secondary daughter ions. The resolution of the secondary scan was 17,500, the collision energy used was 30%, 40% and 50%, and the dynamic exclusion time was set to 3 seconds. High-purity nitrogen was used as sheath gas and auxiliary gas. The sheath gas pressure and auxiliary gas pressure were 35psi and 15arb, respectively. The spray voltage was +3000V. The ion transfer tube temperature was 350°C, and the H-ESI heating temperature was 300°C. An electrospray ion source was used.

[0064] Example 5

[0065] This embodiment relates to a method for analyzing and detecting the residual amount of chlorthiazide in rat feces, which specifically comprises the following steps:

[0066] S1 Pre-treatment of stool samples

[0067] (1) Grind the freeze-dried feces thoroughly, weigh 0.2 g and add 2 mL of acetonitrile. Oscillate the mixture at 250 rpm for 30 min in an oscillator, then centrifuge at 1300 g for 10 min in a high-speed centrifuge to obtain a supernatant. Repeat the above steps 3 times and combine the supernatants.

[0068] (2) First, activate the HLB SPE (60 mg / 3 mL) column with 3 mL of Milli-Q water and 3 mL of acetonitrile, then pass the supernatant obtained in step (1) through the column and collect the effluent.

[0069] (3) The solution obtained in step (2) was nitrogen-blown to 0.5 mL under a gentle nitrogen flow (purity: 99%). The obtained solution was then centrifuged at 9500 g for 5 min, and the supernatant was removed for testing.

[0070] S2 Instrumental Analysis Method

[0071] The target compound chlorothioline was quantitatively analyzed using a liquid chromatography Ultimate 3000 tandem orbitrap high-resolution mass spectrometer Q-Exactive (Liquid Chromatogram-Quadrupole-Orbitrap High-Resolution Mass Spectrometer, ThermoFisher Scientific, USA). The chromatographic column used was Hypersil Gold TM C18 chromatographic column (100 mm × 2.1 mm × 3 μm; Thermo Scientific, USA), pre-column: Hypersil Gold TM C18 (10mm×2.1mm×3μm). The mobile phases were Milli-Q water (A) and acetonitrile (B) containing 0.1% formic acid, respectively. The mobile phase gradient program was 10% B from 0 to 1 min, changed to 100% B within 5 min and maintained for 4 min, and changed to 10% B within 2 min and maintained for 4 min. The injection volume was 5μL and the column temperature was 40°C. The mass spectrometer obtained full scan mass spectral data in positive ionization mode with a scan range of 100-750Da and a resolution of 140,000. After each full scan, the top 10 ions with the strongest response values ​​were fully scanned for secondary daughter ions. The resolution of the secondary scan was 17,500, the collision energy used was 30%, 40% and 50%, and the dynamic exclusion time was set to 3 seconds. High-purity nitrogen was used as sheath gas and auxiliary gas. The sheath gas pressure and auxiliary gas pressure were 35psi and 15arb, respectively. The spray voltage was +3000V. The ion transfer tube temperature was 350°C, and the H-ESI heating temperature was 300°C. An electrospray ion source was used.

[0072] The recovery rate was calculated using the pre-spiking and post-spiking methods. Specifically, the samples in Group 1 were unspiked samples; the samples used in Group 2 were the same as those in Group 1, except that a known amount (25 ng) of standard substances were added to the samples to be tested before the pretreatment operation, which was called the pre-spiking sample; the samples used in Group 3 were the same as those in Groups 1 and 2, except that after the pretreatment operation, the same amount of standard substances as in Group 2 were added, which was called the post-spiking sample. The recovery rate can be calculated by comparing the peak areas measured for the pre-spiked and post-spiked samples after deducting the background concentration, and the calculation formula is as follows:

[0073] Recovery rate = (group 2 - group 1) / (group 3 - group 1) × 100%

[0074] The following are the calculation results of the recovery rate of chlorthiazide in rat plasma, tissues and organs, urine and feces in Examples 2-5.

[0075] Table 1 Recovery rates of pretreatment methods for plasma, tissues, urine and feces

[0076] Matrix Recovery rate (%) plasma 73.5±2.0 Tissues and organs 85.0±3.5 Urine 87.3±3.9 stool 75.7±5.4

[0077] It can be seen that the recovery rate of plasma samples is 73.5±2.0%, the recovery rate of tissue and organ samples is 85.0±3.5%, the recovery rate of urine samples is 87.3±3.9%, and the recovery rate of feces samples is 75.7±5.4%. The present invention has the advantages of high sensitivity, good stability and simple operation.

[0078] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A method for analyzing and detecting the residue of a new type of neonicotinoid pesticide in mammalian plasma, tissues and organs, urine, and feces, characterized in that: The following steps are involved: S1: Pretreatment of mammalian plasma / tissue / urine / feces samples; Among them, the pretreatment method of mammalian plasma is: After thawing the mammalian plasma sample, transfer the plasma sample to a centrifuge tube, add acetonitrile to precipitate protein, vortex and shake, and then centrifuge to obtain a supernatant; The supernatant is concentrated and purified by using an HLB SPE solid phase extraction column. First, the HLB SPE solid phase extraction column is activated with Milli-Q water and an organic solvent. Then, the supernatant is passed through the HLB SPE solid phase extraction column and then the HLB SPE solid phase extraction column is eluted with an organic solvent, and the eluate is collected. The eluate is concentrated by nitrogen blowing under a nitrogen flow, and then the volume is fixed with a solvent. The resulting solution is then centrifuged and the supernatant is removed for testing on an instrument. The pre-treatment method of mammalian tissues and organs is: Weigh the homogenized tissue and organ samples and add an organic solvent, shake and centrifuge to obtain a supernatant; The supernatant was concentrated and purified using an HLB SPE solid phase extraction column. First, the HLB SPE solid phase extraction column was activated with Milli-Q water and an organic solvent, and then the supernatant was passed through the HLB SPE solid phase extraction column and the effluent was collected; The collected liquid was concentrated by nitrogen blowing under a nitrogen flow, and then the volume was fixed with a solvent. The resulting solution was then centrifuged and the supernatant was removed for testing on an instrument. The pretreatment method of mammalian urine is: After the mammalian urine sample is thawed, the urine sample is transferred to an HLB SPE solid phase extraction column for concentration and purification. First, the HLB SPE solid phase extraction column is activated with an organic solvent and Milli-Q water. Then, the urine sample is passed through the HLB SPE solid phase extraction column and then the HLB SPE solid phase extraction column is eluted with an organic solvent, and the eluate is collected. The eluate is then blown to near dryness under a nitrogen stream, and then the volume is fixed with a solvent. The resulting solution is then centrifuged, and the supernatant is removed for testing on an instrument. The pre-treatment methods for mammal feces are: After freeze-drying, the feces of mammals are fully ground, the feces sample is weighed and an organic solvent is added, and after shaking and centrifugation, a supernatant is obtained; The supernatant was concentrated and purified using an HLB SPE solid phase extraction column. First, the HLB SPE solid phase extraction column was activated with Milli-Q water and an organic solvent, and then the supernatant was passed through the HLB SPE solid phase extraction column and the effluent was collected; The collected liquid was concentrated by nitrogen blowing under a nitrogen flow, and then the volume was fixed with a solvent. The resulting solution was then centrifuged and the supernatant was removed for testing on an instrument. S2: The processed samples are tested on the machine. The specific instrument analysis method is as follows: The target compounds were quantitatively analyzed using liquid chromatography Ultimate 3000 tandem orbitrap high-resolution mass spectrometry Q-Exactive.

2. The analysis and detection method according to claim 1, characterized in that: In S2, the chromatographic column used is HypersilGold TM C18 column, pre-column is Hypersil Gold TM C18, the mobile phases were Milli-Q water containing 0.1% formic acid and acetonitrile, respectively. The mass spectrometer obtained full scan mass spectral data in positive ionization mode with a scan range of 100-750Da and a resolution of 140,000. High-purity nitrogen was used as sheath gas and auxiliary gas. The sheath gas pressure and auxiliary gas pressure were 35psi and 15arb, respectively, and the spray voltage was +3000V. The ion transfer tube temperature was 350°C, the H-ESI heating temperature was 300°C, and an electrospray ion source was used.

3. The analysis and detection method according to claim 1, characterized in that: The mammal is one of a mouse, a cat, a dog, a cow, a sheep, a rabbit, a donkey and a horse.

4. The analysis and detection method according to claim 1, characterized in that: The freeze-drying time for mammal feces is 5-7 days.

5. The analysis and detection method according to claim 1, characterized in that: The organic solvent used in the solid phase extraction process was one of acetonitrile, acetonitrile containing 0.1% formic acid, and acetonitrile: dichloromethane with a volume ratio of 3:

7.

6. The analysis and detection method according to claim 1, characterized in that: The solvent used for volume adjustment was acetonitrile.

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