Search

CN-121994969-A - Analysis method and system for detecting multi-type antibiotic residues in edible parts of vegetables

CN121994969ACN 121994969 ACN121994969 ACN 121994969ACN-121994969-A

Abstract

The invention relates to the technical field of food safety detection, and particularly provides an analysis method and an analysis system for detecting various antibiotic residues in edible parts of vegetables. The method aims at the problem that the prior art is difficult to detect various antibiotics in vegetables efficiently and accurately at the same time, realizes synchronous and efficient extraction and purification of sulfonamides, quinolones and tetracycline antibiotics by optimizing a sample pretreatment flow, combines specific ultra-high performance liquid chromatography-tandem high resolution mass spectrometry analysis conditions, and utilizes a pre-constructed standard compound database to carry out intelligent comparison, thereby completing rapid screening and accurate quantification of various target antibiotics in complex vegetable matrixes. The invention provides a complete solution, has the advantages of universality, high efficiency, high detection flux, good accuracy and the like in pretreatment, and provides a reliable technical means for daily monitoring and risk assessment of antibiotic residues in vegetables.

Inventors

  • ZHANG QING
  • XIA XINGHUI
  • CHEN LISHU
  • LI KAIXUAN

Assignees

  • 北京师范大学

Dates

Publication Date
20260508
Application Date
20260311

Claims (10)

  1. 1. An analytical method for detecting antibiotic residues of various types in edible parts of vegetables, comprising the steps of: S1, preprocessing a vegetable sample to obtain a detection liquid to be analyzed; S2, analyzing the detection liquid to be analyzed by adopting an ultra-high performance liquid chromatography-tandem high resolution mass spectrometer to obtain mass spectrum information of compounds in the detection liquid, wherein the mass spectrum information comprises retention time, fragment ion information and parent ion mass information; s3, comparing mass spectrum information of the compounds in the detection liquid with a preset standard antibiotic compound information database, and identifying and quantifying sulfonamide, quinolone and tetracycline antibiotics in the vegetable sample; The preset standard antibiotic compound information database comprises standard mass spectrum information of 52 target antibiotics, and the 52 antibiotics comprise 26 sulfonamide antibiotics, 20 quinolone antibiotics and 6 tetracycline antibiotics.
  2. 2. The method according to claim 1, wherein the step S1 comprises: s11, crushing and preprocessing a vegetable sample; S12, adding an isotope internal standard mixed solution and an extracting solution into the pretreated sample, extracting, and combining the extracting supernatants; s13, concentrating and redissolving the combined extracting supernatant; And S14, enriching and purifying the target compound in the complex solution by adopting a solid phase extraction column, and eluting to obtain the detection liquid to be analyzed.
  3. 3. The method according to claim 2, wherein the pretreatment in step S11 comprises adding Na 2 EDTA, naCl and anhydrous Na 2 SO 4 to the vegetable sample; wherein, every 2g vegetable samples added with 0.2g Na 2 EDTA,0.1g NaCl and 0.2g anhydrous Na 2 SO 4 .
  4. 4. The method according to claim 2, wherein in the step S12, the extracting solution is a mixed solution of methanol, acetonitrile and hydrochloric acid in a volume ratio of 48:48:4.
  5. 5. The method according to claim 2, wherein in step S12, the isotopically internal standard mixed solution comprises sulfamethazine- 13 C 6 , sulfamonomethoxine- 13 C 6 , trimethoprim-d 3 , sulfamethazine-d 4 , sulfamethazine-d 4 , ofloxacin-d 3 and doxycycline-d 3 .
  6. 6. The method according to claim 2, wherein in step S14, the solid phase extraction column is an HLB solid phase extraction column; the enrichment and purification comprises the steps of activating the HLB solid phase extraction column by using methanol and ultrapure water in sequence, loading a complex solution, leaching by using ultrapure water, and eluting a target compound by using methanol after the complex solution is pumped out.
  7. 7. The method according to claim 1, wherein in the step S2, the ultra performance liquid chromatography is performed by using a C18 column, mobile phase a is an aqueous solution containing 1mM formic acid, mobile phase B is a methanol solution containing 1mM formic acid, and gradient elution is performed; The gradient elution procedure was: 0min, mobile phase A90% and mobile phase B10%; 2min, mobile phase A80%, mobile phase B20%; 8min, mobile phase A68%, mobile phase B32%; 12min, mobile phase A65%, mobile phase B35%; 17min, mobile phase A35%, mobile phase B65%; 20min, mobile phase A30%, mobile phase B70%; 21min, mobile phase A5%, mobile phase B95%; 24min, mobile phase A5%, mobile phase B95%; 25min, mobile phase A90% and mobile phase B10%; 29min, mobile phase A90% and mobile phase B10%.
  8. 8. The method according to claim 1, wherein in step S2, the tandem high resolution mass spectrometry employs electrospray ionization source, positive ion mode, multi-reaction monitoring scan mode.
  9. 9. The method according to claim 1, wherein the step S3 specifically includes: S31, matching the parent ion mass information of the compounds in the detection liquid with the parent ion mass information of each standard antibiotic in a database, and screening out suspected target compounds within an error range of +/-5 ppm; S32, comparing and confirming the retention time and fragment ion information of the suspected target compound with the retention time and fragment ion information of corresponding standard antibiotics in a database; And S33, quantitatively analyzing the target compound confirmed to exist according to the peak area ratio of the target compound to the corresponding isotope internal standard and a standard curve by adopting an internal standard method.
  10. 10. A system for carrying out the analytical method for detecting antibiotic residues of multiple classes in edible vegetable portions according to claim 1, comprising: the pretreatment module is used for carrying out pretreatment on the vegetable sample to obtain detection liquid to be analyzed; The mass spectrum analysis module is used for analyzing the detection liquid to be analyzed by adopting an ultra-high performance liquid chromatography-tandem high resolution mass spectrometer to obtain mass spectrum information of the compounds in the detection liquid; The data processing module is used for comparing mass spectrum information of the compounds in the detection liquid with a preset standard antibiotic compound information database, and identifying and quantifying sulfonamide, quinolone and tetracycline antibiotics in the vegetable sample; the preset standard antibiotic compound information database comprises standard mass spectrum information of 52 target antibiotics.

Description

Analysis method and system for detecting multi-type antibiotic residues in edible parts of vegetables Technical Field The invention relates to the technical field of environmental pollutant and food safety analysis and detection, in particular to an analysis method and an analysis system for detecting multi-class antibiotic residues in edible parts of vegetables. Background Antibiotics are used in large quantities worldwide as important drugs for preventing and treating infectious diseases of humans and livestock. The production and use of antibiotics in China are large, and a large number of antibiotics which are not completely metabolized enter the agricultural environment through livestock and poultry manure and other ways. These manure containing antibiotic residues is applied to farmlands as organic fertilizer or is input through irrigation water, so that antibiotics are continuously accumulated in the soil of the farmlands, and become a new pollutant of great concern. Vegetable planting systems are an important link for migration of antibiotics from soil to the food chain. Antibiotics have certain durability in the environment, and can be absorbed by vegetable root systems and transported to edible parts. Humans, on a long-term basis, ingest vegetables containing low doses of antibiotics, may interfere with the homeostasis of the intestinal flora and may accelerate the development and spread of bacterial resistance, potentially threatening public health. Therefore, various antibiotics are listed in a new pollutant list which is mainly controlled in China, and the enhancement of monitoring of the antibiotics in agricultural products is important. Currently, methods for detecting antibiotics in environments and agricultural products are mostly focused on water, soil or substrate sludge, and are generally directed to only a single class (e.g., sulfonamides or quinolones) or a limited number (typically within 20) of antibiotics. However, the matrix of the edible parts of vegetables is extremely complex, and contains a large amount of endogenous interfering substances such as chlorophyll, pigments, organic acids, saccharides, oils and fats. These substances not only severely interfere with the detection signal of the instrument for trace antibiotics, but also make development of a pretreatment method capable of extracting multi-class antibiotics from complex vegetable substrates simultaneously and efficiently pose a great challenge due to the great difference in the binding capacity of the substances with antibiotics of different polarities and chemical properties. In addition, the difference of the tissue structure and chemical composition of different types of vegetables (such as lettuce of leaf vegetables, tomatoes of fruits and vegetables and peppers) is remarkable, and the difficulty of establishing a multi-class antibiotic residue analysis method with strong universality is further increased. Therefore, the prior art lacks an integrated analysis method capable of rapidly, accurately and simultaneously measuring tens of common antibiotic residues in various vegetables. This technical bottleneck results in that we cannot fully evaluate the mixed contamination status of antibiotics in vegetables and their dietary exposure risk. The development of an analysis method which is suitable for complex vegetable matrixes and can synchronously detect three major antibiotics such as sulfonamides, quinolones, tetracyclines and the like is a key technical problem to be solved in food safety supervision and risk assessment. Disclosure of Invention The invention aims to solve or improve the technical problem that the multi-class and multi-antibiotic residues in vegetables cannot be synchronously, efficiently and accurately detected in the related technology. In order to solve the technical problems, the technical scheme provided by the application is as follows: the invention provides an analysis method for detecting multi-class antibiotic residues in edible parts of vegetables, which comprises the following steps: S1, preprocessing a vegetable sample to obtain a detection liquid to be analyzed; S2, analyzing the detection liquid to be analyzed by adopting an ultra-high performance liquid chromatography-tandem high resolution mass spectrometer to obtain mass spectrum information of compounds in the detection liquid, wherein the mass spectrum information comprises retention time, fragment ion information and parent ion mass information; s3, comparing mass spectrum information of the compounds in the detection liquid with a preset standard antibiotic compound information database, and identifying and quantifying sulfonamide, quinolone and tetracycline antibiotics in the vegetable sample; The preset standard antibiotic compound information database comprises standard mass spectrum information of 52 target antibiotics, and the 52 antibiotics comprise 26 sulfonamide antibiotics, 20 quinolone antibiotics and 6 tetracycline