Abstract
In this work, the preparation of an aptamer-based coating for stir bar sorptive extraction (SBSE) aimed at the selective clean-up and preconcentration of the allergenic milk protein β-lactoglobulin (β-LG) is described. To this end, a thiol-terminated aptamer was grafted onto a vinylized polytetrafluoroethylene stir bar via “thiol-ene” click chemistry. The aptamer affinity coating was characterized and several parameters that affect SBSE efficiency were evaluated. The optimized aptamer-based SBSE method combined with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) analysis provided suitable linearity (0.25–50 µg·mL−1), satisfactory limit of detection (LOD) (0.08 µg·mL−1 β-LG), and relative standard deviations below 5.2 % among the extraction units, which could be reused for 10 extraction cycles and were stable for at least one month. The applicability of the developed method was successfully demonstrated by analyzing low levels of β-LG in different dairy-free food samples (i.e. a 100 % cocoa dark chocolate, a hydrolyzed hypoallergenic infant formula, and a white bread) and in white breads with declared and undeclared contents of milk (84 %<recoveries < 93 %).
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