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

Sufang Fan; Junmei Ma; Chunsheng Li; Yanbo Wang; Wen Zeng; Qiang Li; Jinru Zhou; Liming Wang; Yi Wang; Yan Zhang

doi:10.3389/fnut.2022.848294

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

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

Authors

Sufang Fan¹, Junmei Ma^{1

2}, Chunsheng Li³, Yanbo Wang⁴, Wen Zeng⁵, Qiang Li¹, Jinru Zhou⁴, Liming Wang¹, Yi Wang⁵, Yan Zhang^{1

2}

Affiliations

¹ Hebei Food Safety Key Laboratory, Key Laboratory of Special Food Supervision Technology for State Market Regulation, Hebei Food Inspection and Research Institute, Shijiazhuang, China.
² Hebei Key Laboratory of Forensic Medicine, College of Forensic Medicine, Hebei Medical University, Shijiazhuang, China.
³ Biology Institute of Hebei Academy of Science, Shijiazhuang, China.
⁴ School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, China.
⁵ Department of Chemical Engineering, Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Tsinghua University, Beijing, China.

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.