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Method for simultaneously detecting AZA1, AZA2 and AZA3 in bivalve aquatic products

A technology for aquatic products and bivalves, which is applied in the field of determination of various protopolyformic acid shellfish toxins in bivalve aquatic products, and can solve the problem of time-consuming, inability to determine the specific components of various toxins, and sample processing operations cumbersome and other problems, to achieve the effect of improving the efficiency of detection and analysis

Inactive Publication Date: 2013-10-09
孙兴权 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Mouse bioassay (MBA) is a common method for detecting shellfish toxins in the past, but because of its inability to determine the specific components of various toxins, the specificity, sensitivity and accuracy are not good, and the reproducibility is poor. It has been replaced by the liquid chromatography-tandem mass spectrometry (LC-MS / MS) method adopted by the European Union in 2011
At present, the detection of AZAs shellfish toxins is mostly limited to AZA1, and the single AZA1 determination result cannot be used as the basis for judging whether AZAs in bivalve aquatic products exceeds the standard, and the established detection method still has cumbersome sample processing operations and takes a long time. question

Method used

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  • Method for simultaneously detecting AZA1, AZA2 and AZA3 in bivalve aquatic products
  • Method for simultaneously detecting AZA1, AZA2 and AZA3 in bivalve aquatic products
  • Method for simultaneously detecting AZA1, AZA2 and AZA3 in bivalve aquatic products

Examples

Experimental program
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Embodiment 1

[0049] Extraction: Take scallops, oysters, and variegated clams, which are representative bivalve aquatic products, as the research objects. Weigh 100g of the edible parts of the three samples, wash and homogenize at 9500r / min for 5min for sample preparation. Weigh 2 g of the homogeneous sample to be tested (accurate to 0.01 g) and place it in a 50 mL plastic centrifuge tube with a stopper, add 8 mL of 80% (V / V) methanol aqueous solution, vortex extract for 2 min, and extract at 8000 r / min at 4 °C Centrifuge for 5 minutes, transfer the supernatant to another centrifuge tube, add 8 mL of 80% (V / V) methanol aqueous solution to the residue and re-extract once, combine the supernatant; add 5 mL of n-hexane to the supernatant, and vortex at 1500 r / min Extract for 1 min, remove the n-hexane layer after static layering, and obtain extract I; add 15 mL of chloroform to extract I, vortex extract at 1500 r / min for 2 min, centrifuge at 8000 r / min at 4 °C for 2 min, remove the supernatant,...

Embodiment 2

[0065] Embodiment 2. Application to actual sample detection

[0066] Using the detection method established in Example 1, 50 actual samples of bivalve aquatic products sold in the market were tested, and no AZAs shellfish toxins were detected. It is reported that AZA1 has been detected in shellfish samples in Dalian sea area with a content of 99.6pg / g, but a single AZA1 measurement result cannot be used as a basis for judging whether AZAs in bivalve aquatic products exceed the standard. According to the European Union's detection requirement that the total amount of AZAs does not exceed 160 μg / kg, the present invention does not pursue a lower detection limit too much, and can fully meet the requirements of AZAs in bivalve aquatic products under the detection limit of LOQ=1.5 μg / kg. The need for routine testing and monitoring of shellfish toxins.

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Abstract

The invention discloses a method for simultaneously detecting AZA1, AZA2 and AZA3 in bivalve aquatic products. According to the method, liquid-liquid extraction and hybrid anion exchange reversed phase adsorption solid-phase extraction and purification are utilized to perform sample treatment on scallops, oysters, variegated clams and other bivalve aquatic products, and an analysis and detection method for simultaneously determining three main azaspiracid shellfish poisons AZA1, AZA2 and AZA3 in a sample through a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method is established; and furthermore, the average recovery rate of AZAs is more than 80%, and the relative standard deviation (RSD) is less than 12%. The method disclosed by the invention has the characteristics of high speed, high efficiency, sensitivity and accuracy.

Description

technical field [0001] The present invention relates to a method for determining multiple protopolyformic acid shellfish toxins in bivalve aquatic products, in particular to a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of three main protoxins in bivalve aquatic products. Methods of polyforminoid shellfish toxins AZA1, AZA2 and AZA3. Background technique [0002] Azaspiracids (AZAs) shellfish toxins are a class of fat-soluble polyether biotoxins produced by Properidininm crassipes. The carbon skeleton consists of 40 carbon atoms, and there are 20 stereoisomers in the molecule. It has a structural center and 9 rings, has a unique 6,5,6-triple spiro ring and cyclic amine structure (see structural formula 1), and belongs to the aminohelicic acid shellfish toxin. AZA1 is the most common and the highest proportion of AZAs shellfish toxins, AZA2 and AZA3 are the 8-methyl and 22-demethyl derivatives of AZA1, respectively, and these thre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 孙兴权郑秋月李一尘曹际娟
Owner 孙兴权
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