title: Development of a Method for Crustacean Allergens Using Liquid Chromatography/Tandem Mass Spectrometry.
abstract: 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. 22ADTLEQQNK30, 92IQLLEEDLER101, 113LAEASQAADESER125, 134SLSDEER140, 153FLAEEADR160, and 190IVELEEELR198 of tropomyosin and 152VSSTLSSLEGELK164 and 217TFLVWVNEEDHLR229 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 (r2>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.
authors.affiliation: Gifu Prefectural Research Institute for Health and Environmental Science, 1-1 Naka-Fudougaoka, Kakamigahara, 504-0838, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nagai
authors.forename: Tomiaki
authors.initials: T
authors.lastname: Minatani
authors.forename: Kotaro
authors.initials: K
authors.lastname: Goto
mesh_terms: D000485:Allergens / Q000032:analysis*; D020816:Amino Acid Motifs; D000818:Animals; D001122:Arginine Kinase / Q000032:analysis; D060829:Arthropod Proteins / Q000032:analysis*; D002853:Chromatography, Liquid / Q000379:methods*; D003445:Crustacea / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D005504:Food Analysis / Q000379:methods*; D005512:Food Hypersensitivity / Q000517:prevention & control; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D059748:Proteolysis; D016415:Sequence Alignment; D012758:Shellfish / Q000032:analysis*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014335:Tropomyosin / Q000032:analysis
chemical_list: D000485:Allergens; D060829:Arthropod Proteins; D010446:Peptide Fragments; D014335:Tropomyosin; D001122:Arginine Kinase