Quantification of crustacean tropomyosin in foods using high‐performance liquid chromatography–tandem mass spectrometry method

Abstract

BACKGROUND Allergic reactions to crustacean products have been increasing owing to the rising consumption. Tropomyosin (TM) is the main crustacean allergen; it has a coiled‐coil structure, which shows stability to various food processing methods. Crustacean processed products have been used in several food products, thereby causing greater difficulties in detecting TM in these products. We aimed to develop an assay based on high‐performance liquid chromatography–tandem mass spectrometry for the accurate and reproducible quantification of crustacean TM in foods. RESULTS The three peptides IQLLEEDLER, LAEASQAADESER, and IVELEEELR were selected as peptide markers, and the peptide IVELEEELR was selected as the quantitative marker. Extraction conditions and enzymatic digestion conditions were completely optimized. The extraction solution of Tris–hydrochloric acid buffer (50 mmol L−1, pH 7.4) containing 1 mol L−1 potassium chloride and the enzymatic treatment at 1:15 ratio (enzyme/protein, m/m) for 13 h showed excellent efficiency. The method exhibited a good linear relationship, with the qualified coefficient of determination (R2 = 0.9994) in the wide range of 1 to 1000 μg L−1. The accuracy was validated based on spiked recovery at three spiking levels (12.5, 25.0, and 50.0 μg kg−1, TM/matrix) in blank matrices that included chicken sausages, beef balls, and egg–milk biscuits. The recoveries ranged from 91% to 109% with qualified relative standard deviations <15% with the limit of quantification (of 1.6 mg kg−1, TM/matrix). CONCLUSION This new approach can be used for the qualitative and quantitative detection of crustacean TM in various food matrices. © 2021 Society of Chemical Industry.

Multisyringe Flow Injection Analysis of Tropomyosin Allergens in Shellfish Samples

“…These two methods do not measure the same compounds. Tropomyosin is composed of two polypeptide chains, with five peptides found after digestion [ 37 ]. Among them, the peptide with the IVELEEELR sequence has been shown to be the most sensitive and was thus chosen to be a marker of tropomyosin [ 37 ].…”

“…Tropomyosin is composed of two polypeptide chains, with five peptides found after digestion [ 37 ]. Among them, the peptide with the IVELEEELR sequence has been shown to be the most sensitive and was thus chosen to be a marker of tropomyosin [ 37 ]. The alternative method proposed in this paper found glutamic acid to be the most prevalent amino acid in that peptide, and it was thus chosen to be a marker of tropomyosin.…”

“…The SDS-PAGE bands between 34 and 40 kDa ( Appendix A ) were destained and digested with trypsin, according to the method designed by Abel Rahman et al [ 26 ], before injection into the LC/MS system. Tropomyosin quantification was based on IVELEEELR (sequence) peptide determination, as suggested by Wang et al [ 37 ].…”

A comprehensive overview of emerging processing techniques and detection methods for seafood allergens

“…J. Wang et al. (2021) also developed an assay based on high‐performance liquid chromatography‐tandem mass spectrometry (LC–HRMS) to detect crustacean TM qualitatively and quantitatively. They found that this approach demonstrated high accuracy and reproducibility (recovery range of 91−109%, standard deviations < 15%, limit of quantification of 1.6 mg/kg) in various food matrices (chicken sausages, beef balls, and egg–milk biscuits).…”

An overview of tropomyosin as an important seafood allergen: Structure, cross‐reactivity, epitopes, allergenicity, and processing modifications

“…There are various types of allergic reactions, which can be divided into Type‐I: anaphylaxis, Type‐II: cytotoxic, Type‐III: immune complex, and Type‐IV: late‐onset (Maker et al., 2019; Trautmann & Stoevesandt, 2019). In individuals susceptible to TM, IgE‐mediated allergic reaction (Type‐I allergic reaction) is the most common (Jianhua Wang et al., 2021). The IgE‐mediated hypersensitivity reaction mainly covers two steps based on seafood allergy.…”