The determination of an allergen protein , β-casein in nano-LC system and its intraction with some phenolic compounds

In this study, i) preparation of a new 9-Anthracenemethyl methacrylate (ANM) monolithic nanocolumn with hydrophobic character for reversed-phase nano-liquid chromatography (nano-LC), ii) new preparation for the allergenic protein β-casein (β-CN) developing a determination method and iii) applying it to milk samples and examining the cojugate structures formed as a result of its interaction with some phenolic compounds. In this context, a new hydrophobic monolithic nanocolumn based on Anthracenemethacrylate (ANM), a new approach for the determination of β-CN, was prepared and characterized. First, monolithic column solutions containing ANM and divinylbenzene (DVB) were prepared, cyclohexanol and dodecanol were used as porogenic solvents, and a monolithic- nano column was prepared by in-situ polymerization method. ANM monolithic column was prepared with an inner diameter of 100 µm and a length of 20 cm, and its structural morphology was examined with SEM images. In addition, chromatographic characterization of the column was performed using homologous alkylbenzenes in the nano-LC system. The β-CN determination method was developed in the nano-LC system using the characterized monolithic column, and the LOD value was found to be 0.2 mg/mL. The developed method was applied to milk samples and quantification was made. ın addition, conjugated structures formed by the allergenic protein β-CN and some phenolics were examined