Development of a liquid chromatography-tandem mass spectrometry method for simultaneous quantification of hen's egg white allergens Gal d 1-4 in fresh and processed eggs

Hen's egg white allergens, namely Gal d 1-4, cause food allergies worldwide and their intake must be strictly controlled by allergic individuals. However, an efficient method for quantifying these allergens is currently unavailable. We aimed to develop an LC-MS/MS method for simultaneous Gal d 1-4 quantification. Purified Gal d 1-4 proteins were trypsin-digested and the resulting peptides used in LC-MS/MS analysis. The limits of quantification were 9.77-39.1 ng/mL. The Gal d 1-4 recovery in fresh and processed eggs was 68.3-121.3%, and intra- and interassay coefficients of variation were 1.5-15.7% and 2.4-38.1%, respectively, indicating high sensitivity, accuracy, and reproducibility. In addition, the high specificity of this method was confirmed by testing 27 other foods. This newly developed method could provide reliable information to the industrial food and clinical fields, facilitating improved quality of life for individuals with egg allergies.