Development of a Qualitative Analytical Method for Milk, Egg, Buckwheat, and Peanut Allergens Using LC-MS/MS

LC-MS/MS was used to develop a qualitative analytical method for milk, egg, buckwheat, and peanut allergens. The measurement conditions and extraction and trypsin digestion process for protein pretreatment were optimized. A sample spiked with each allergen was analyzed to evaluate the performance of the analytical method, including the extraction process. Repeatability and intralaboratory precision met the target values for all allergens, confirming the stability of the analytical method. Furthermore, the S/N ratio, retention time, and peptide ratio demonstrated the usefulness of the method as a qualitative test. Moreover, for milk and egg allergens, the equivalence of the quantitative values calculated using the developed method to those of ELISA was confirmed; however, this equivalence could not be confirmed for commercially available processed foods and some quality control samples. For buckwheat and peanut allergens, the equivalence of the quantitative values to those of ELISA could not be confirmed in the spiked sample. These findings demonstrate that this analytical method is useful as a rapid and simple qualitative test. It is also an excellent alternative to ELISA as it produces less waste and reduces environmental burden.