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Method for detecting 15 antibiotics in fish body

A technology for antibiotics and fish body, applied in the field of organic pollutant residue detection, can solve the problems of limited types and quantities of antibiotics, limited recovery rate, and many interferences, and achieve the effects of short extraction time, stable recovery rate, and low dosage.

Pending Publication Date: 2022-07-08
SHANGHAI ACADEMY OF ENVIRONMENTAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no complete and reliable analysis method for the separation, enrichment and co-detection of multiple antibiotics in aquatic products. The existing methods generally have problems such as instability, high interference, limited types and quantities of antibiotics, and limited recovery.

Method used

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  • Method for detecting 15 antibiotics in fish body
  • Method for detecting 15 antibiotics in fish body
  • Method for detecting 15 antibiotics in fish body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] 1. Sample pretreatment

[0107] Wash the collected fish samples, absorb the surface moisture with gauze, dissect the back muscles of the fish, separate the fish skin, add the same quality of water to the fish samples, and then homogenize them. The treated samples are quickly placed at -20°C. Freeze until further extraction.

[0108] Weigh 5.00g of homogenized fish samples into a 30mL glass centrifuge tube, and add 100ng of sulfamethoxazole-D respectively. 4 , Trimethoprim-D 3 , Norfloxacin-D 5 , Furazolidone-D 4 , tetracycline-D 6 , Olaquindox-D 4 and florfenicol-D 3 The 7 internal standards were thoroughly mixed and allowed to stand in the dark for 10 min. Then add 10 mL of acetonitrile to the centrifuge tube and mix by vortex for 1 min. Acetonitrile can denature the protein and release the drug bound to it. Then add 10 mL of phosphate buffer with pH=3 and mix by vortex for 1 min. The buffer provides the target substance. In a relatively stable pH environment, ...

Embodiment 2

[0139] The matrix of fish meat samples is complex and contains compounds such as protein, fat, carbohydrate, inorganic salt and water. These sample matrices will affect the extraction efficiency of the analyte, cause errors in the detection results, and also contaminate the instrument, which directly affects the detection efficiency and accuracy. Therefore, in the analysis of veterinary drug residues, the pretreatment technology of the sample is very critical, and the main purpose is to separate the target compound from the sample matrix and remove interfering impurities. At the same time, there are many kinds of target antibiotics, and the physicochemical properties vary greatly. In order to maximize the extraction efficiency of various antibiotics in fish meat, the present invention adopts a single factor experiment to study the extraction effect of three different extraction solutions on the target substances.

[0140] Homogenized fish samples were obtained according to ste...

Embodiment 3

[0143] According to step 2 of Example 1, the mixed standard solution of 15 antibiotics was diluted with 70% methanol-water solution into a series of concentration gradients (1.37, 4.12, 12.35, 37.04, 111.11, 333.33, 500 μg·L -1 ), the internal standard is 100.0 μg·L -1 The concentration of 1 was added to the standard product, and the internal standard method was used to draw a 7-point standard curve with the concentration ratio of the component to the internal standard as the abscissa, and the response value (response value=target peak area / internal standard peak area) as the ordinate to draw a 7-point standard curve to obtain Standard curve and correlation coefficient (R 2 ). Meanwhile, the limit of detection and limit of quantification were determined by the signal-to-noise ratio (S / N) method, the limit of detection (LOD) was estimated by 3 times the signal-to-noise ratio, and the limit of quantification (LOQ) was estimated by 10 times the signal-to-noise ratio. The standa...

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Abstract

The invention provides a method for detecting 15 antibiotics in a fish body, which comprises the following steps: sequentially adding an internal standard substance and an extraction solution into a fish meat sample, carrying out ultrasonic extraction, adding the obtained extraction solution into a solid-phase extraction column, enriching, eluting, degreasing, and determining the obtained sample solution by adopting ultra-high performance liquid chromatography-tandem mass spectrometry, 15 antibiotics in the sample solution are determined according to the retention time, an internal standard curve method is adopted for quantification, and the content of the 15 antibiotics in the sample solution is determined. According to the method for detecting the 15 antibiotics in the fish body, provided by the invention, 7 types of 15 antibiotics in fish meat can be rapidly, efficiently, sensitively and accurately detected at the same time, the detection efficiency is greatly improved, and the method has the advantages of high recovery rate, high sensitivity, high stability, low detection limit, more real and reliable detection result and the like.

Description

technical field [0001] The invention belongs to the technical field of organic pollutant residue detection, and relates to a method for detecting 15 kinds of antibiotics in fish, in particular to an ultrasonic extraction-solid-phase extraction combined with ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS / MS) ) method to detect the content of 15 antibiotics in fish. Background technique [0002] In recent years, my country's aquaculture industry has developed rapidly, and large-scale and intensive aquaculture has become the trend of aquaculture, and the frequency of aquatic animal diseases is also increasing. In order to prevent and control aquatic product diseases and accelerate the growth and reproduction of aquatic products, antibiotics are widely and massively used in aquaculture. The ionic form of antibiotics has a strong affinity for proteins and DNA, and the input antibiotics can accumulate in aquatic products through bioaccumulation. A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/72G01N30/8634G01N2030/045Y02A40/81
Inventor 胡双庆沈根祥张洪昌李贞金郭文宏朱英李雨薇
Owner SHANGHAI ACADEMY OF ENVIRONMENTAL SCIENCES
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