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

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...

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Published inFrontiers in nutrition (Lausanne) Vol. 9; p. 848294
Main Authors Fan, Sufang, Ma, Junmei, Li, Chunsheng, Wang, Yanbo, Zeng, Wen, Li, Qiang, Zhou, Jinru, Wang, Liming, Wang, Yi, Zhang, Yan
Format Journal Article
LanguageEnglish
Published Switzerland Frontiers Media S.A 03.03.2022
Subjects
immunoaffinity purification
isotope-label
liquid chromatography-tandem mass spectrometry
Nutrition
signature peptide
tropomyosin
liquid chromatography-tandem mass spectrometry
immunoaffinity purification
tropomyosin
isotope-label
signature peptide
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Summary: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.
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Reviewed by: Xizhi Shi, Ningbo University, China; Wentong Xue, China Agricultural University, China
Edited by: Biao Yuan, China Pharmaceutical University, China
This article was submitted to Food Chemistry, a section of the journal Frontiers in Nutrition
These authors have contributed equally to this work and share first authorship
ISSN:2296-861X
2296-861X
DOI:10.3389/fnut.2022.848294