Abstract
The quantification of allergens in food including baked food matrices is of great interest. The aim of the present study was to describe a non-immunologic method to quantify bovine β-casein using ultra-performance liquid chromatography tandem triple quadrupole mass spectrometry (UPLC-TQ-MS/MS) in multiple reaction monitoring (MRM) mode. Eight of 10 theoretical peptides from β-casein after tryptic digestion were compared and MRM methods were developed to determine five signature peptides. The peptide VLPVPQK was selected as the signature peptide for bovine β-casein because of the high sensitivity. A stable isotope-labelled internal standard was designed to adjust the instability of sample pre-treatment and ionisation caused by matrix effect. Using the present suspension digestion method, the native and denatured β-casein could be digested to release the signature peptide at the maximum extent. The UPLC-TQ-MS/MS method developed based on a tryptic signature peptide led to a reliable determination of bovine β-casein allergen in baked food matrices at a low quantitation level down to 500 μg kg–1 with a satisfactory accuracy (< 8.9%) and recovery (98.8% ± 2.6% to 106.7% ± 3.0%).
Highlights
About 2% of the general population suffers from food allergies (Ring et al 2001)
An in silico tryptic digestion of bovine β-casein was performed with the PeptideMass tool provided by UniProt
Ten of the 16 theoretical digestion products could be used as the potential candidates for a signature peptide (Table 1)
Summary
About 2% of the general population suffers from food allergies (Ring et al 2001). Allergy to cow’s milk is considered as one of the most common food allergies (Bock & Atkins 1990). β-Casein is one of the major allergens in cow’s milk proteins (IUIS Allergen Nomenclature 2014). Allergy to cow’s milk is considered as one of the most common food allergies (Bock & Atkins 1990). There were some cases in which allergenic ingredients were unrecognised or not declared on the product labels (hidden allergens) (Anibarro et al 2007; Van Hengel 2007), which put food-allergic consumers at great risk. In order to prevent these accidents from happening, food manufacturers and food testing laboratories are responsible for ensuring that products are free of allergens or labelled . The detection methods of food allergens include PCR and ELISA as the traditional techniques and MS-based methods as emerging techniques (Poms et al 2004b; Monaci et al 2006; Johnson et al 2010)
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