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Immunoaffinity Column Purification-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Detection of 3-Methyl-quinoxaline-2-carboxylic Acid in Fish and Shrimp

A technique of ultra-high performance liquid phase and tandem mass spectrometry, which is applied in the detection field of 3-methyl-quinoxaline-2-carboxylic acid and achieves the effect of good reproducibility and high sensitivity

Active Publication Date: 2019-12-03
MARINE FISHERIES RES INST OF ZHEJIANG
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Immunoaffinity column (IAC) is based on antigen and antibody reaction, has the advantages of strong specificity, high enrichment rate and good purification efficiency, and has been successfully applied in the purification and enrichment of aflatoxin, tetrodotoxin and other substances, while There is no literature report on the purification of MQCA in aquatic products by immunoaffinity column

Method used

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  • Immunoaffinity Column Purification-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Detection of 3-Methyl-quinoxaline-2-carboxylic Acid in Fish and Shrimp
  • Immunoaffinity Column Purification-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Detection of 3-Methyl-quinoxaline-2-carboxylic Acid in Fish and Shrimp
  • Immunoaffinity Column Purification-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Detection of 3-Methyl-quinoxaline-2-carboxylic Acid in Fish and Shrimp

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Embodiment

[0021] 1.1 Materials and reagents

[0022] MQCA standard (purity ≥ 98.0%, Dr. Ehrenstorfer, Germany); Waters Oasis HLB solid phase extraction column (60mg / 3mL); Waters Oasis MAX mixed anion exchange column (60mg / 3mL); MQCA immunoaffinity column (column Volume 1mL, Beijing Huaan Maike Biotechnology Co., Ltd.) methanol, acetonitrile (chromatographically pure, Merck, Germany), formic acid (analytical pure, Sigma, USA), acetic acid, HCl, Na 2 HPO 4 12H 2 O, NaH 2 PO 4 2H 2 O, NaCl, NaOH (analytical grade, Shanghai Sinopharm Group). The experimental water was Milli-Q to prepare ultrapure water (resistivity ≥ 18.2MΩ.cm).

[0023] MQCA standard solution: Accurately weigh 5.00 mg of MQCA standard product, dissolve it in methanol and dilute to 50 mL, the solution concentration is 100 μg / mL, and store at -20°C in the dark. The intermediate solution and the working solution were gradually diluted to the required concentration with 0.1% formic acid-acetonitrile solution (9:1, V / V)....

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Abstract

The invention discloses a method for detecting MQCA (3-methyl-2-quinoxalinecarboxylic acid) in fish and prawns by adopting IAC-UPLC-MS / MS (immunoaffinity column purification-ultrahigh performance liquid chromatography-tandem mass spectrometry). According to the method, a well homogenized sample is subjected to acidolysis in an HCl solution with concentration of 2 mol / L, then pH of an extracting solution is regulated to 7-8, after IAC enrichment and purification, LC-MS / MS (liquid chromatography-tandem mass spectrometry) is adopted for determination, and an external standard method is adopted for quantification. 0.1% formic acid aqueous solution-methanol is taken as a mobile phase, gradient elution separation is performed, and monitoring is performed under conditions of electrospray ionization, positive ions and a multi-reaction monitoring mode. The method is good in reproducibility, high in sensitivity and suitable for actual determination of MQCA in aquatic products.

Description

technical field [0001] The invention relates to a method for detecting 3-methyl-quinoxaline-2-carboxylic acid, in particular to the detection of 3-methyl-quinoxaline in fish and shrimp by immunoaffinity column purification-ultra-high performance liquid chromatography-tandem mass spectrometry -2-Carboxylic acid method. Background technique [0002] 3-Methylquinoxaline-2-carboxylic acid (MQCA) is a residual marker of quinoxaline drug olaquindox, and also a metabolite of quinocetone and acetylmethaquine. Studies have shown that olaquindox has obvious teratogenic effects on most animals, and it also has potential teratogenic, mutagenic, and carcinogenic effects on humans. Therefore, it is prohibited or restricted to be used as a feed additive worldwide. Lalaquindox is rapidly metabolized in the body, while MQCA has a long residence time in the body, and the relationship between its content and total residues is stable. It is one of the labeled residues identified by the Codex ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 李佩佩张小军严忠雍陈思
Owner MARINE FISHERIES RES INST OF ZHEJIANG