A method for detecting multiple veterinary drug residues in animal-derived food

Abstract

The present invention relates to the technical field of food detection, and specifically to a method for detecting various veterinary drug residues in animal-derived foods, comprising the following steps: extraction: using acetonitrile solution as the main extraction solvent to extract animal-derived foods to obtain an extract; purification: purifying the extract based on FaVEx column rapid extraction and purification technology to obtain a purified solution; detection: detecting various veterinary drug residues in the purified solution based on ultra-high performance liquid chromatography tandem mass spectrometry technology UPLC‑MS / MS. The present invention improves the extraction and purification time, reduces the amount of organic solvents used, responds to the call for green chemistry, saves costs, shortens the waiting time for veterinary residue detection, and improves sampling efficiency and production capacity.

Description

Technical Field

[0001] The invention relates to the technical field of food detection, and in particular to a method for detecting multiple veterinary drug residues in animal-derived food. Background Art

[0002] Food safety is an important issue concerning people's livelihood. With the improvement of people's living standards, the consumption of animal-derived products is increasing, and the safety of animal-derived foods is one of the focuses of social attention. Veterinary drugs are widely used to treat or prevent animal diseases, and play an important role in reducing animal morbidity and mortality, promoting animal growth, and improving the quality of animal products. However, the irrational use of drugs and drug abuse cause drug residues in animals, resulting in the detection of banned drugs and veterinary drug residues exceeding national standards. The phenomenon has increased year by year, bringing hidden risks to food safety. Drug residues may have direct or potential harmful effects on consumer health, including allergic reactions, increased risk of drug resistance, and "three-cause" reactions.

[0003] To this end, countries and organizations such as China, Japan, the European Union, and the International Food Commission have established residue limits for drugs in animal-derived foods. my country's veterinary drug residue limit standards list 267 types of veterinary drugs in three categories, of which 104 have established maximum residue limit standards, 9 are allowed for therapeutic use, but must not be detected in animal foods, and the remaining drugs not included in the standard may also pose a threat to human health. Due to the large number of derivatives of various veterinary drugs and the large differences in the chemical properties of different types of veterinary drugs, the detection efficiency is low, making it difficult to meet the needs of veterinary drug multi-residue detection.

[0004] In summary, for animal-derived foods, establishing a set of rapid screening methods for veterinary drug residues with wide coverage, high sensitivity, rapidity and accuracy has important practical application value. Summary of the invention

[0005] In order to solve the above problems, the present invention provides a method for detecting multiple veterinary drug residues in animal-derived foods, which is used to detect multiple veterinary drug residues in animal-derived foods simply, quickly and efficiently.

[0006] In order to achieve the above object, the technical solution of the present invention is as follows: A method for detecting multiple veterinary drug residues in animal-derived food comprises the following steps:

[0007] Extraction: using acetonitrile solution as the main extraction solvent to extract the animal-derived food to obtain an extract;

[0008] Purification: Based on the FaVEx column rapid extraction purification technology, the extract is purified to obtain a purified solution;

[0009] Detection: Based on ultra-high performance liquid chromatography tandem mass spectrometry technology UPLC-MS / MS, various veterinary drug residues in the purified liquid are detected.

[0010] Furthermore, in the extraction, the selection of extraction solvents: comparative analysis of the extraction effects of seven extraction solutions on the target object, including acetonitrile solution, 0.5% formic acid acetonitrile, 1% formic acid acetonitrile, 2% formic acid acetonitrile, 0.5% acetic acid acetonitrile, 1% acetic acid acetonitrile and 2% acetic acid acetonitrile.

[0011] Furthermore, in the purification process, the selection of purification methods: QuEChERS method, FaVEx column rapid extraction purification technology and national standard method purification technology were selected for experimental analysis, and the key steps of each purification method were optimized to remove fat, protein and organic acid in the matrix.

[0012] Furthermore, during the detection, the mass spectrometry conditions are optimized: according to the molecular weight and charging mode, the standard solution is directly injected into the ion source through a needle or pump, and the primary mass spectrometry analysis is performed to obtain the molecular ion peak, determine the parent ion of each target, and optimize the declustering voltage; the parent ion is scanned by secondary mass spectrometry to obtain fragment ion information, and 2 to 3 characteristic ions with higher response and stability are selected.

[0013] Furthermore, the cone voltage and collision energy corresponding to each compound are optimized, the best response fragment ions are determined, and the mass spectrometry parameters of the target compound are determined.

[0014] Furthermore, the slope ratio of the standard curve was selected to evaluate the matrix effect ME, and the relative ratio of the two was calculated by the following formula to evaluate the matrix effect ME:

[0015]

[0016] Where A and B represent the peak areas of the analyte in pure solvent and matrix solution, respectively.

[0017] Furthermore, the detection method was validated by analyzing the linearity, detection limit, quantification limit, accuracy, precision and stability of the method.

[0018] The above scheme has the following beneficial effects:

[0019] 1. The present invention utilizes the FaVEx column rapid extraction and purification technology to rapidly extract and purify multiple veterinary drug residues in animal-derived foods. Compared with conventional methods and the currently widely used QuEChERS method, the extraction and purification time is increased, the amount of organic solvents used is reduced, and the call for green chemistry is responded to. At the same time, costs are saved, the waiting time for veterinary residue detection is shortened, and the sampling efficiency and production capacity are improved.

[0020] 2. The present invention applies the analytical method of ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS / MS) combined with FaVEx column rapid extraction and purification technology to other animal-derived products, and determines the simplicity, rapidity, efficiency, economy and applicability of the method.

[0021] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a flow chart of an embodiment of a method for detecting multiple veterinary drug residues in animal-derived food of the present invention;

[0023] Figure 2 A schematic diagram of the selection and analysis of extraction solvents in an embodiment of the method for detecting multiple veterinary drug residues in animal-derived food of the present invention;

[0024] Figure 3 A schematic diagram of purification mode selection analysis for an embodiment of a method for detecting multiple veterinary drug residues in animal-derived food of the present invention;

[0025] Figure 4 This is a schematic diagram of the total ion flow of veterinary drugs in an embodiment of the method for detecting multiple veterinary drug residues in animal-derived food of the present invention. DETAILED DESCRIPTION

[0026] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] The following is further described in detail through specific implementation methods:

[0028] Implementation example Figure 1 As shown: A method for detecting multiple veterinary drug residues in animal-derived food comprises the following steps:

[0029] Extraction: Use acetonitrile solution as the main extraction solvent to extract animal-derived food and obtain an extract. Among them, the selection of extraction solvent: acetonitrile and methanol are commonly used extraction agents in veterinary drug residue detection, but the protein content in the methanol solution in the preliminary experiment was high and the solvent was turbid, which was not conducive to sample purification and detection. Therefore, acetonitrile was selected as the main extraction solvent. And compare and analyze the extraction effects of seven extraction solutions on the target object, including acetonitrile solution, 0.5% formic acid acetonitrile, 1% formic acid acetonitrile, 2% formic acid acetonitrile, 0.5% acetic acid acetonitrile, 1% acetic acid acetonitrile and 2% acetic acid acetonitrile. As shown in the attached Figure 2 shown.

[0030] Purification: Based on the FaVEx column rapid extraction purification technology, the extract is purified to obtain a purified solution. Among them, animal-derived foods are rich in lipids, proteins and other compounds. Compared with agricultural products, the matrix is ​​more complex, there are more interfering substances, and the content of veterinary drugs is very small, which makes separation and purification more difficult. Therefore, the choice of purification method: QuEChERS method, FaVEx column rapid extraction purification technology and national standard method purification technology are selected for experimental analysis, and the key steps of each purification method are optimized to remove interfering substances such as fat, protein and organic acid in the matrix, thereby reducing the matrix effect in the instrumental analysis process, and obtaining practical and usable purification methods, such as the attached Figure 3 shown.

[0031] Detection: Based on the ultra-high performance liquid chromatography tandem mass spectrometry technology UPLC-MS / MS, various veterinary drug residues in the purified liquid are detected. Among them, the mass spectrometry conditions are optimized: according to the molecular weight and charging mode, the standard solution is directly injected into the ion source through a needle or pump, and the primary mass spectrometry analysis is performed to obtain the molecular ion peak, determine the parent ion of each target, and optimize the declustering voltage; the parent ion is scanned by secondary mass spectrometry to obtain fragment ion information, and 2 to 3 characteristic ions with high response and stability are selected, such as the attached Figure 4 shown.

[0032] Since most of the tested compounds have strong signal values ​​under ESI+, the cone voltage and collision energy corresponding to each compound are optimized to determine the best response fragment ion and the mass spectrometry parameters of the target compound.

[0033] Matrix effect investigation: The contents of water, protein, fat, minerals, etc. in different matrices are all different, so the response intensity will be different when performing method analysis. That is, the matrix effect is different. Therefore, it is necessary to evaluate the matrix effect to ensure the accuracy and feasibility of the method. Therefore, the slope ratio of the standard curve is selected to evaluate the matrix effect ME, and the relative ratio of the two is calculated by the following formula to evaluate the matrix effect ME:

[0034]

[0035] Where A and B represent the peak area (slope) of the analyte in pure solvent and matrix solution, respectively.

[0036] Validation of the detection method: The feasibility of this method was verified by analyzing its linearity, detection limit, quantification limit, accuracy, precision and stability.

[0037] To sum up the above, this embodiment uses FaVEx column rapid extraction and purification technology to quickly extract and purify multiple veterinary drug residues in animal-derived foods, thereby improving the extraction and purification time, reducing the use of organic solvents, responding to the call for green chemistry, and saving costs. It shortens the waiting time for veterinary residue detection and improves sampling efficiency and production capacity.

[0038] 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 detecting multiple veterinary drug residues in animal-derived food, characterized in that: The steps include: Extraction: using acetonitrile solution as the main extraction solvent to extract the animal-derived food to obtain an extract; Purification: Based on the FaVEx column rapid extraction purification technology, the extract is purified to obtain a purified solution; Detection: Based on ultra-high performance liquid chromatography tandem mass spectrometry technology UPLC-MS / MS, various veterinary drug residues in the purified liquid are detected.

2. The method for detecting multiple veterinary drug residues in animal-derived food according to claim 1, characterized in that: During the extraction, the selection of extraction solvents: comparative analysis of the extraction effects of seven extraction solutions on the target object, including acetonitrile solution, 0.5% formic acid acetonitrile, 1% formic acid acetonitrile, 2% formic acid acetonitrile, 0.5% acetic acid acetonitrile, 1% acetic acid acetonitrile and 2% acetic acid acetonitrile.

3. The method for detecting multiple veterinary drug residues in animal-derived food according to claim 1, characterized in that: During purification, the choice of purification method: QuEChERS method, FaVEx column rapid extraction purification technology and national standard method purification technology were selected for experimental analysis, and the key steps of each purification method were optimized to remove fat, protein and organic acid in the matrix.

4. The method for detecting multiple veterinary drug residues in animal-derived food according to claim 1, characterized in that: During the detection, the mass spectrometry conditions are optimized: according to the molecular weight and charging mode, the standard solution is directly injected into the ion source through a needle or pump, and the primary mass spectrometry analysis is performed to obtain the molecular ion peak, determine the parent ion of each target, and optimize the declustering voltage; the parent ion is scanned by secondary mass spectrometry to obtain fragment ion information, and 2 to 3 characteristic ions with higher response and stability are selected.

5. The method for detecting multiple veterinary drug residues in animal-derived food according to claim 4, characterized in that: Optimize the cone voltage and collision energy corresponding to each compound, determine the best response fragment ion, and determine the mass spectrometry parameters of the target compound.

6. The method for detecting multiple veterinary drug residues in animal-derived food according to claim 1, characterized in that: The slope ratio of the standard curve was selected to evaluate the matrix effect ME, and the relative ratio of the two was calculated by the following formula to evaluate the matrix effect ME: Where A and B represent the peak areas of the analyte in pure solvent and matrix solution, respectively.

7. The method for detecting multiple veterinary drug residues in animal-derived food according to claim 1, characterized in that: Validation of the detection method: Analyze the linearity, detection limit, quantification limit, accuracy, precision and stability of this method.

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