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. 2022 Mar 3:9:848294.
doi: 10.3389/fnut.2022.848294. eCollection 2022.

Determination of Tropomyosin in Shrimp and Crab by Liquid Chromatography-Tandem Mass Spectrometry Based on Immunoaffinity Purification

Sufang Fan  1 , Junmei Ma  1   2 , Chunsheng Li  3 , Yanbo Wang  4 , Wen Zeng  5 , Qiang Li  1 , Jinru Zhou  4 , Liming Wang  1 , Yi Wang  5 , Yan Zhang  1   2
Affiliations

Determination of Tropomyosin in Shrimp and Crab by Liquid Chromatography-Tandem Mass Spectrometry Based on Immunoaffinity Purification

Sufang Fan et al. Front Nutr. .

Abstract

A UPLC-MS/MS method was developed for the detection of tropomyosin (TM) in shrimp and crab. After simple extraction, the samples were purified by immunoaffinity column and then digested by trypsin. The obtained sample was separated by Easy-nLC 1000-Q Exactive. The obtained spectrums were analyzed by Thermo Proteome Discoverer 1.4 software and then ANIQLVEK with high sensitivity was selected as the quantitative signature peptide. Isotope-labeled internal standard was used in the quantitative analysis. The method showed good linearity in the range of 5-5,000 μg/L with a limit of quantification (LOQ) of 0.1 mg/kg. The average recoveries were 77.22-95.66% with RSDs ≤ 9.97%, and the matrix effects were between 88.53 and 112.60%. This method could be used for rapid screening and quantitative analysis of TM in shrimp and crab. Thus, it could provide technical support for self-testing of TM by food manufacturers and promote further improvement of allergen labeling in China.

Keywords: immunoaffinity purification; isotope-label; liquid chromatography-tandem mass spectrometry; signature peptide; tropomyosin.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Chromatographic-mass spectrograms of signature peptides and internal standard of tropomyosin.
See this image and copyright information in PMC

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