Method for detecting residual quantity of cyanuric acid substances in pet food

Abstract

The invention relates to the technical field of food detection, and particularly discloses a method for detecting residual quantity of cyanuric acid substances in pet food, which comprises the following steps: (1) weighing 0.01 g of cyanuric acid substance standard substance in a 10mL volumetric flask, and preparing 1000mu g / mL of standard stock solution by using methanol water; transferring 1.00 mL of the standard stock solution into a 100 mL volumetric flask, and diluting with methanol to a scale to obtain a standard solution; (2) weighing 0.5 g of a pet food sample, putting the pet food sample into a 10mL centrifugal tube, adding a mixed solution of a functional additive, acetonitrile and water into the centrifugal tube until the scale and vortex are reached, then carrying out ultrasonic treatment, taking supernate, and filtering the supernate through an organic filter membrane to obtain a sample solution; and (3) performing LC-MS / MS analysis on the standard solution and the sample solution, and calculating the residual quantity of the cyanuric acid substances according to a formula. The method for detecting the residual quantity of the cyanuric acid substances in the pet food gives consideration to accuracy, stability and convenience in the actual application process, and has a remarkable application effect.

Description

Technical Field

[0001] The present application relates to the technical field of food detection, and more specifically, to a method for detecting the residual amount of cyanuric acid substances in pet food. Background Art

[0002] Melamine is a triazine-type nitrogen-containing heterocyclic organic compound and an important organic chemical raw material. It is widely used in the production of plastics, coatings, adhesives, food packaging materials, disinfectants, fertilizers and pesticides. It may enter food from the environment, food packaging materials, etc.

[0003] Melamine is generally relatively stable, but it may release cyanide at high temperatures. In a strong acid or alkaline environment, melamine will hydrolyze, and the amino group will be gradually replaced by hydroxyl groups to generate melamine homologues, mainly cyanuric acid, cyanuric acid monoamide, and cyanuric acid diamide. If the above-mentioned cyanuric acid substances appear in pet food, it will affect the health of pets, and relevant tests on such substances are required.

[0004] At present, the methods for detecting cyanuric acid substances mainly include high performance liquid chromatography-diode array method, high performance liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, etc. Before the above methods are applied, pet food needs to be processed to reduce interference and obtain more accurate detection results. Usually, it is first treated with a mixture of acetonitrile and water, and then treated with acetonitrile saturated n-hexane. This two-step treatment method can effectively precipitate protein and fat, effectively reduce interference, and thus ensure accurate detection of cyanuric acid residues in pet food.

[0005] Regarding the above-mentioned related technologies, the inventors believe that although the above-mentioned processing of pet food can effectively reduce interference, the two-step processing method is not only relatively complicated in actual operation, but also easy to cause artificial loss of sample solution during the operation. The stability and convenience of the overall application still need to be improved.

[0006] Therefore, it is urgent to propose a solution to solve the above technical problems. Summary of the invention

[0007] In order to ensure accurate measurement of cyanuric acid substances in pet food and improve the stability and convenience of the entire detection application, the present application provides a method for detecting the residual amount of cyanuric acid substances in pet food.

[0008] The present application provides a method for detecting the residual amount of cyanuric acid substances in pet food, which adopts the following technical scheme: A method for detecting the residual amount of cyanuric acid substances in pet food comprises the following steps: (1) Preparation of standard solution: weigh 0.01 g of cyanuric acid standard into a 10 mL volumetric flask, and prepare a 1000 ug / mL standard stock solution with methanol and water; then transfer 1.00 mL of the standard stock solution to a 100 mL volumetric flask, and dilute to the mark with methanol to obtain a standard solution; (2) Extraction of sample solution: Weigh 0.5 g of pet food sample and place it in a 10 mL centrifuge tube. Add a mixed solution of functional additives, acetonitrile and water to the centrifuge tube to the mark. Vortex and then perform ultrasonic treatment. Take the supernatant and filter it through an organic filter membrane to obtain a sample solution. The mass mixing ratio of the functional additive, acetonitrile and water is (0.4-0.6):(3-4):1; The functional additive is composed of diethylamine and trichloroacetic acid, and the mass mixing ratio of diethylamine to trichloroacetic acid is 1:(8-10); (3) Perform LC-MS / MS analysis on the standard solution and the sample solution, and calculate the residual amount of cyanuric acid substances according to the following formula; X = A × C × V / (A0 × m); X-residue of cyanuric acid in pet food samples, mg / kg; A0-peak area of ​​cyanuric acid substances in standard solution; 1. Peak area of ​​cyanuric acid in sample solution; C-Concentration of cyanuric acid in standard solution, μg / mL; V-volume of sample solution, mL; m-mass of pet food sample, g; The cyanuric acid-based substance is any one of cyanuric acid, ammeline and ammelide.

[0009] By adopting the above technical scheme, in the extraction and preparation of the sample solution, through the use of functional additives, with acetonitrile-water solution, the protein and fat in the pet food sample can be effectively precipitated in a one-time mixing process, so that the extraction and purification are completed at the same time, reducing interference, making the entire process more convenient and simple, less prone to inaccurate measurement results caused by human factors, and more stable overall application. Among them, the functional additives are composed of diethylamine and trichloroacetic acid. Diethylamine is beneficial to improving the extraction rate of cyanuric acid substances, and the amino group in the diethylamine molecule can combine with the fatty acid in the oil to form acetamide, and its unique molecular structure makes it have a stripping effect; trichloroacetic acid can protonate the carboxyl group on the amino acid of the protein, which has the effect of assisting the precipitation of protein; diethylamine and trichloroacetic acid can play an excellent synergistic role with acetonitrile, so that the protein molecules and lipid molecules in the pet food sample can be effectively separated and processed, and the dissolution and acquisition of cyanuric acid substances can be accelerated, so that the cyanuric acid substances in the pet food can be accurately measured later. It can be seen that the detection method of cyanuric acid residues in pet food can take into account accuracy, stability and convenience in actual application, and has significant application effect.

[0010] Preferably, the functional additive consists of diethylamine and trichloroacetic acid, and the mass mixing ratio of diethylamine to trichloroacetic acid is 1:8.5.

[0011] By adopting the above technical scheme, when diethylamine and trichloroacetic acid in the above weight ratio are used in combination, the balance between their applications is better, and the coordination effect with acetonitrile is better, so that in the extraction process of the sample solution, the protein and fat removal effect is better, and the interference in the subsequent detection process is also lower, so that the final result of the residual amount of cyanuric acid substances in the pet food sample is more accurate.

[0012] Preferably, the mass mixing ratio of the functional additive, acetonitrile and water is 0.5:3.5:1.

[0013] By adopting the above technical scheme, the functional additive, acetonitrile and water are applied to the pet food sample in the above mass mixing ratio, so that the corresponding effect of the combination of the functional additive and acetonitrile is relatively excellent, and a one-time mixing treatment can not only effectively eliminate the interference of protein and fat, but also completely extract cyanuric acid substances, thereby making the detection method of the residual amount of cyanuric acid substances in pet food have better accuracy, stability and convenience.

[0014] Preferably, in step (2), during the ultrasonic treatment, the temperature is controlled at 20-28°C.

[0015] By adopting the above technical solution, during the extraction process of the sample solution, ultrasonic extraction will generate heat. If the temperature is too high, it may affect the stability of the sample and the extraction effect. The selection of the above temperature is conducive to obtaining a better extraction effect of cyanuric acid substances.

[0016] Preferably, in step (2), the pore size of the organic filter membrane is 0.2-0.3 μm.

[0017] By adopting the above technical solution, the organic filter membrane of the above specifications can achieve better filtering and separation effects on impurities in the extraction solution, thereby obtaining a relatively pure sample solution, ensuring the accuracy of subsequent LC-MS / MS analysis, and ultimately obtaining a more accurate residual amount of cyanuric acid substances in pet food.

[0018] Preferably, in step (3), during the LC-MS / MS analysis, the mobile phase comprises phase A and phase B, phase A is a 10% ammonium acetate aqueous solution, phase B is acetonitrile, and the volume ratio of the 10% ammonium acetate aqueous solution to acetonitrile is 1:(4-9).

[0019] By adopting the above technical solution, the above-selected phase A and phase B, and the selection of a suitable volume ratio between the two, a stable and better elution effect can be achieved for the standard solution and the sample solution in the LC-MS / MS analysis, which is beneficial to ensure the accuracy of the spectrum. When the subsequent calculation is performed according to the formula, a more accurate residual amount of cyanuric acid substances in pet food can be obtained.

[0020] Preferably, the volume ratio of the 10% ammonium acetate aqueous solution to acetonitrile is 1:6.

[0021] By adopting the above technical solution, after the above volume ratio of 10% ammonium acetate aqueous solution and acetonitrile are used, the corresponding effect brought about is relatively stable, which is conducive to making the residual amount of cyanuric acid substances in the obtained pet food more accurate.

[0022] Preferably, the pet food sample is any one of dry pet food, semi-dry pet food and pet liquid food.

[0023] By adopting the above technical scheme, the above types of pet food samples are all suitable for the detection method of the residual amount of cyanuric acid substances in this application, and a relatively accurate residual amount of cyanuric acid substances can be obtained. The overall application range is wide and the application effect is outstanding.

[0024] In summary, this application has the following beneficial effects: The present application adopts diethylamine and trichloroacetic acid as functional auxiliary agents, and cooperates with acetonitrile-water solution to extract pet food samples, which not only has an excellent extraction rate of cyanuric acid substances, but also can effectively precipitate proteins and fats in pet food samples, so that extraction and purification are completed at the same time, reducing interference; in this way, the detection method of the residual amount of cyanuric acid substances in pet food takes into account accuracy, stability and convenience in actual application, and has a significant application effect. DETAILED DESCRIPTION

[0025] The present application is further described in detail below in conjunction with embodiments and comparative examples.

[0026] Unless otherwise specified, the raw materials used in the preparation examples, embodiments and comparative examples of the present application are commercially available.

[0027] Standard substances: Cyanuric acid (CAS No.: 108-80-5, molecular formula: C3H6N6) ≥ 99%; Ammelide (CAS No.: 645-93-2, molecular formula: C3H4N4O2) ≥ 99%; Ammeline (CAS No.: 645-92-1, molecular formula: C3H5N5O) ≥ 99%. Example

[0028] Example 1 A method for detecting the residual amount of cyanuric acid substances in pet food comprises the following steps: (1) Preparation of standard solution: weigh 0.01 g of cyanuric acid standard into a 10 mL volumetric flask, and prepare a 1000 ug / mL standard stock solution with methanol and water; then transfer 1.00 mL of the standard stock solution to a 100 mL volumetric flask, and dilute to the mark with methanol to obtain a standard solution; (2) Extraction of sample solution: Weigh 0.5 g of pet food sample and place it in a 10 mL centrifuge tube. Add a mixed solution of functional additives, acetonitrile and water to the scale. Vortex for 3 min and then perform ultrasonic treatment for 30 min. Take the supernatant and filter it through an organic filter membrane to obtain a sample solution. (3) LC-MS / MS analysis: The standard solution and the sample solution were subjected to LC-MS / MS analysis, and the residual amount of cyanuric acid substances was calculated according to the following formula; X = A × C × V / (A0 × m); X-residue of cyanuric acid in pet food samples, mg / kg; A0-peak area of ​​cyanuric acid substances in standard solution; 2. Peak area of ​​cyanuric acid in the sample solution; C-Concentration of cyanuric acid in standard solution, μg / mL; V-volume of sample solution, mL; m-mass of pet food sample, g; Note: In the above step (2), the mass mixing ratio of the functional additive, acetonitrile and water is 0.5:3.5:1; the functional additive is composed of diethylamine and trichloroacetic acid, and the mass mixing ratio of diethylamine and trichloroacetic acid is 1:8.5; during the ultrasonic treatment, the temperature is controlled to be 24°C; the pore size of the organic filter membrane is 0.25 μm; in step (3), during the LC-MS / MS analysis, the mobile phase includes phase A and phase B, phase A is 10% ammonium acetate aqueous solution, phase B is acetonitrile, and the volume ratio of 10% ammonium acetate aqueous solution to acetonitrile is 1:6.

[0029] The cyanuric acid substance can be cyanuric acid, ammeline or ammelide, and the corresponding chromatographic conditions and mass spectrometric conditions in LC-MS / MS analysis are as follows: Chromatographic conditions: Chromatographic column: UPLC BEH Amide 1.7μm 2.1х100mm; Mobile phase: Item A: 10% ammonium acetate aqueous solution; Item B: acetonitrile; Column oven temperature: 40°C; Injection volume: 5 μL; Flow rate: 0.4 mL / min; The gradient is as follows: Time Module Command Value 0.15min Pumps PumpB Conc. 87 2.00min Pumps PumpB Conc. 55 2.50min Pumps PumpB Conc. 40 5.50min Pumps PumpB Conc. 40 5.51min Pumps PumpB Conc. 87 7.5 controller stop Mass spectrometry conditions: Interface Tem: 350℃; DL Tem: 200℃; Heat Block Tem: 200℃; Example 2 A method for detecting residual amounts of cyanuric acid substances in pet food, which differs from Example 1 in that the mass mixing ratio of the functional additive, acetonitrile and water is 0.4:3:1.

[0030] Example 3 A method for detecting residual amounts of cyanuric acid substances in pet food, which differs from Example 1 in that the mass mixing ratio of the functional additive, acetonitrile and water is 0.6:4:1.

[0031] Example 4 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that in Example 1 in that the functional additive is composed of diethylamine and trichloroacetic acid in a mass mixing ratio of 1:8.

[0032] Example 5 A method for detecting residual amounts of cyanuric acid substances in pet food, which differs from Example 1 in that the functional additive is composed of diethylamine and trichloroacetic acid in a mass mixing ratio of 1:10.

[0033] Example 6 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that in Example 1 in that the functional additive is composed of diethylamine and trichloroacetic acid in a mass mixing ratio of 1:9.

[0034] Example 7 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that of Example 1 in that in step (2), during the ultrasonic treatment, the temperature is controlled at 20°C.

[0035] Example 8 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that in Example 1 in that in step (2), during the ultrasonic treatment, the temperature is controlled at 28°C.

[0036] Example 9 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that in Example 1 in that, in step (2), the pore size of the organic filter membrane is 0.2 μm.

[0037] Example 10 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that in Example 1 in that, in step (2), the pore size of the organic filter membrane is 0.3 μm.

[0038] Embodiment 11 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that of Example 1 in that the volume ratio of the 10% ammonium acetate aqueous solution to acetonitrile is 1:4.

[0039] Example 12 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that of Example 1 in that the volume ratio of the 10% ammonium acetate aqueous solution to acetonitrile is 1:9.

[0040] Embodiment 13 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that of Example 1 in that the volume ratio of 10% ammonium acetate aqueous solution to acetonitrile is 1:6.5.

[0041] Embodiment 14 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that of Example 1 in that in step (2), during the ultrasonic treatment, the temperature is controlled at 19°C.

[0042] Embodiment 15 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that in Example 1 in that in step (2), during the ultrasonic treatment, the temperature is controlled at 29°C.

[0043] Example 16 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that of Example 1 in that the volume ratio of 10% ammonium acetate aqueous solution to acetonitrile is 1:3.8.

[0044] Embodiment 17 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that of Example 1 in that the volume ratio of the 10% ammonium acetate aqueous solution to acetonitrile is 1:9.2.

[0045] Embodiment 18 A method for detecting residual amounts of cyanuric acid substances in pet food is different from that in Example 1 in that the functional additive is composed of diethylamine and trichloroacetic acid in a mass mixing ratio of 1:7.8.

[0046] Embodiment 19 A method for detecting residual amounts of cyanuric acid substances in pet food, which differs from Example 1 in that the functional additive is composed of diethylamine and trichloroacetic acid in a mass mixing ratio of 1:10.2.

[0047] Comparative Example Comparative Example 1 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that of Example 1 in that no functional additive is used during the extraction of the sample solution.

[0048] Comparative Example 2 A method for detecting residual amounts of cyanuric acid substances in pet food, which differs from Example 1 in that the functional additive does not contain diethylamine.

[0049] Comparative Example 3 A method for detecting the residual amount of cyanuric acid substances in pet food is different from that in Example 1 in that the functional additive does not contain trichloroacetic acid.

[0050] Performance test samples: 0.25 mg / kg of cyanuric acid, 1.0 mg / kg of ammeline and 0.25 mg / kg of ammeline standards were added to pet food in which cyanuric acid, ammeline and ammeline were not detected. The pet food was purchased from Zhixiao A0501 all-season cat food, and three test samples containing cyanuric acid, ammeline and ammeline were obtained.

[0051] Test method: The detection method of the residual amount of cyanuric acid substances in pet food of Examples 1-19 and Comparative Examples 1-3 was used to detect the test samples containing cyanuric acid, ammeline and ammeline respectively. The recovery rate was calculated according to the residual amount of cyanuric acid substances obtained by different detection methods, that is, recovery rate = actual measurement / added amount, and the corresponding data were recorded in Table 1 below.

[0052] Table 1 Application test results of Examples 1-19 and Comparative Examples 1-3 In combination with Example 1 and Comparative Examples 1-3 and Table 1, it can be seen that in the extraction process of the sample solution, the functional additive composed of diethylamine and trichloroacetic acid can be used to effectively precipitate the protein and fat in the pet food sample, so that extraction and purification are completed at the same time, and excellent cyanuric acid extraction effect is obtained, and then after the detection method is applied, the measured recovery rate is relatively high, indicating that it is more accurate. If diethylamine or trichloroacetic acid is used alone in combination with acetonitrile-water solution, it is found that the recovery rate obtained by the detection method in the actual application process is reduced. Although it is expected that the recovery rate is improved compared to the detection method without using diethylamine and trichloroacetic acid, the improvement effect is extremely limited, and the sum of the improvement effects brought by the use of diethylamine and trichloroacetic acid alone is also far less than the excellent effect brought by the compounding of the two. It can be seen that the application of diethylamine and trichloroacetic acid as a functional additive in the detection method of the residual amount of cyanuric acid in pet food is conducive to taking into account accuracy, stability and convenience, and has a significant application effect.

[0053] It can be seen from Example 1 and Example 2-3 and Table 1 that during the extraction of the sample solution, when the mass mixing ratio of the functional agent, acetonitrile and water is (0.4-0.6):(3-4):1, the detection method can maintain better and more stable accuracy during actual application. Among them, when the mass mixing ratio of the functional agent, acetonitrile and water is 0.5:3.5:1, the detection method for the residual amount of cyanuric acid substances in pet food is applied and tested. The higher the recovery rate obtained, the more accurate the overall application is.

[0054] In combination with Example 1 and Example 4-6 and Table 1, it can be seen that among the functional additives, when the mass mixing ratio of diethylamine and trichloroacetic acid is 1: (8-10), the effect brought by the coordinated application is relatively excellent and stable, and when the mass mixing ratio of diethylamine and trichloroacetic acid is 1: 8.5, the application effect is best; In combination with Examples 18-19 and Table 1, it can be seen that when the mass mixing ratio of diethylamine and trichloroacetic acid exceeds the above range, it is found that the detection method for the residual amount of cyanuric acid substances in pet food samples is significantly reduced after the application of the detection. It can be seen that diethylamine and trichloroacetic acid can only bring excellent compound synergistic effects within a specific mass ratio range.

[0055] Combining Example 1 and Example 7-8 with Table 1, it can be seen that in step (2), during the ultrasonic treatment process, the temperature is controlled to be 20-28°C, so that after the functional auxiliary agent-acetonitrile-water solution is applied, a better extraction effect can be obtained, thereby ensuring the accuracy of the detection of the residual amount of cyanuric acid substances. Combining Examples 14-15 with Table 1, it can be seen that when the ultrasonic temperature exceeds the above range, the extraction effect is deteriorated, and it is difficult to obtain a more accurate detection result.

[0056] It can be seen from Example 1 and Examples 9-10 and Table 1 that in step (2), the pore size of the organic filter membrane is 0.2-0.3 μm, which can obtain a relatively pure sample solution, ensure the accuracy of the subsequent LC-MS / MS analysis, and ultimately obtain a relatively accurate residual amount of cyanuric acid substances in pet food.

[0057] Combining Example 1 and Example 11-13 and Table 1, it can be seen that in the LC-MS / MS analysis process, when the volume ratio of 10% ammonium acetate aqueous solution to acetonitrile is 1:(4-9), a stable and better elution effect is achieved for the standard solution and the sample solution, which is conducive to ensuring the accuracy of the spectrum, and then after calculation by the formula, a more accurate residual amount of cyanuric acid substances in pet food is obtained, and the recovery rate obtained by testing is also relatively high. Combining Example 16-17 and Table 1, it can be seen that when the volume ratio of 10% ammonium acetate aqueous solution to acetonitrile exceeds the above range, it will affect the accurate acquisition of the residual amount of cyanuric acid substances in pet food, and the recovery rate obtained by testing is also significantly reduced, indicating that it has an adverse effect on the accuracy of detection amplification.

[0058] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A method for detecting the residual amount of cyanuric acid substances in pet food, characterized in that: The following steps are involved: (1) Preparation of standard solution: Weigh 0.01 g of cyanuric acid standard into a 10 mL volumetric flask and prepare a 1000 ug / mL standard stock solution with methanol and water; then transfer 1.00 mL of the standard stock solution to a 100 mL volumetric flask and dilute to the mark with methanol to obtain a standard solution; (2) Extraction of sample solution: Weigh 0.5 g of pet food sample and place it in a 10 mL centrifuge tube. Add a mixed solution of functional additives, acetonitrile and water to the centrifuge tube to the mark. Vortex and then perform ultrasonic treatment. Take the supernatant and filter it through an organic filter membrane to obtain a sample solution. The mass mixing ratio of the functional additive, acetonitrile and water is (0.4-0.6): (3-4): 1; The functional additive is composed of diethylamine and trichloroacetic acid, and the mass mixing ratio of diethylamine and trichloroacetic acid is 1: (8-10); (3) LC-MS / MS analysis: The standard solution and the sample solution were subjected to LC-MS / MS analysis, and the residual amount of cyanuric acid substances was calculated according to the following formula; X = A × C × V / (A0 × m); X-residue of cyanuric acid in pet food samples, mg / kg; A0-peak area of ​​cyanuric acid substances in standard solution; Peak area of ​​cyanuric acid substances in the sample solution; C-Concentration of cyanuric acid in standard solution, μg / mL; V-volume of sample solution, mL; m-mass of pet food sample, g; The cyanuric acid-based substance is any one of cyanuric acid, ammeline and ammelide.

2. The method for detecting the residual amount of cyanuric acid substances in pet food according to claim 1, characterized in that: The functional auxiliary agent consists of diethylamine and trichloroacetic acid, and the mass mixing ratio of diethylamine to trichloroacetic acid is 1:8.

5.

3. The method for detecting the residual amount of cyanuric acid substances in pet food according to claim 2, characterized in that: The mass mixing ratio of the functional additive, acetonitrile and water is 0.5:3.5:

1.

4. The method for detecting the residual amount of cyanuric acid substances in pet food according to claim 1, characterized in that: In step (2), during the ultrasonic treatment, the temperature is controlled at 20-28°C.

5. The method for detecting the residual amount of cyanuric acid substances in pet food according to claim 1, characterized in that: In step (2), the pore size of the organic filter membrane is 0.2-0.3 μm.

6. The method for detecting the residual amount of cyanuric acid substances in pet food according to claim 1, characterized in that: In step (3), during the LC-MS / MS analysis, the mobile phase includes phase A and phase B, phase A is a 10% ammonium acetate aqueous solution, phase B is acetonitrile, and the volume ratio of the 10% ammonium acetate aqueous solution to acetonitrile is 1:(4-9).

7. The method for detecting the residual amount of cyanuric acid substances in pet food according to claim 6, characterized in that: The volume ratio of the 10% ammonium acetate aqueous solution to acetonitrile is 1:

6.

8. The method for detecting the residual amount of cyanuric acid substances in pet food according to claim 1, characterized in that: The pet food sample is any one of dry pet food, semi-dry pet food and pet liquid food.

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