Comparison of tropomyosin released peptide and epitope mapping after in vitro digestion from fish (Larimichthys crocea), shrimp (Litopenaeus vannamei) and clam (Ruditapes philippinarum) through SWATH-MS based proteomics

Quantification of Allergic Crustacean Tropomyosin Using Shared Signature Peptides in Processed Foods with a Mass Spectrometry-Based Proteomic Strategy

“…Additionally, it is necessary to employ SIIS to compensate for matrix effects. The use of stable isotope-labeled internal standards such as IVE [L ( 13 C, 15 N)] EEELR helps minimize variations in ionization efficiency. The SIIS also enables monitoring of the mass spectrometry environment to ensure the consistent physical and chemical state of other peptides.…”

“…The standard curve for TM was prepared by adding concentrations of 0.15, 0.75, 1.5, 3, 7.5, 15, and 30 mg/kg (equivalent to 10, 50, 100, 200, 500, 1000, and 2000 ng/mL, respectively) to corresponding samples that did not contain crustacean allergenic TM. An appropriate amount of IVEL ( 13 C, 15 N) EEELR was taken to achieve a concentration of 5 ng/mL, preparing a matrixmatched standard working curve. This process was carried out in parallel with the samples.…”

“…TM standard: purified shrimp tropomyosin was procured from Indoor Biotech (Virginia, USA). The synthetic peptides including LNTATTK, LAEASQAADESER, LAMVEADLER, IVELEEELR, SLEVSEEK, LEDELVNEK, and IVEL ( 13 C, 15 N) EEELR (95% standard) were chemically synthesized by China Peptides (Suzhou, China). Sequencing-grade modified trypsin was obtained from Promega (Madison, WI, USA).…”

“…However, it is accompanied by drawbacks such as the complexity of sample preparation and requires specialized personnel to operate the equipment. − The use of the MS platform has proven to be highly sensitive, allowing for the direct detection of allergenic proteins, which is not achievable using polymerase chain reaction (PCR) technology. Additionally, this method has overcome the limitations of cross-interference commonly encountered in current immunological methods. − In recent years, the development and application of analytical methods based on liquid chromatography tandem mass spectrometry for detecting allergens in complex foodstuffs have been witnessed. − Indeed, while there have been advancements in the development and application of liquid chromatography tandem mass spectrometry for detecting allergens in complex foods, there remains a need for more extensive research on the application of mass spectrometry-based methods for allergen detection in commercially processed foods such as chips and hotpot seasoning.…”