Development of a Method for Crustacean Allergens Using Liquid Chromatography/Tandem Mass Spectrometry

摘要

An LC/MS/MS analysis method was developed for crustacean allergens, tropomyosin, and arginine kinase. A protein extract from shrimp was reduced, alkylated, and digested by trypsin. Peptide spectra were obtained using full scan analysis by LC/MS/MS, and we determined a sequence through a protein search. (22)ADTLEQQNK(30), (92)IQLLEEDLER(101), (113)LAEASQAADESER(125), (SLSDEER140)-S-134, (153)FLAEEADR(160), and (IVELEEELR198)-I-190 of tropomyosin and (192)VSSTLSSLEGELK(164) and (TFLVWVNEEDHLR229)-T-217 of arginine kinase were selected as the specific peptides, and optimal multiple-reaction monitoring conditions were used. The results obtained through the LC/MS/MS analysis correlated well with those using the ELISA method for various crustacean samples (r(2) > 0.9). Moreover, unregulated species, such as krill or insects, which produce positive results in some crustacean ELISA assays, can be differentiated by LC/MS/MS. These findings suggest that LC/MS/MS analysis may be effective for crustacean food allergen analysis.