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Updated: Apr 3, 2026

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
Published on: June 21, 2021
Modification pattern of whey protein depends on oxidizing agents and accelerated storage conditions
Laura Meissner1, Niklas Roeder1, Tobias Demetrowitsch1
1Institute of Human Nutrition and Food Science, Department of Food Technology, University of Kiel, Heinrich-Hecht-Platz 10, 24118 Kiel, Germany.
None:
Oxidative modification sites in whey proteins depend on the oxidizing agent and cannot be described by a single analytical parameter. This study therefore examined how oxidative protein modification differs across oxidizing agents and treatment conditions by evaluating whey proteins at multiple structural levels. Overall protein changes were assessed by RP-HPLC and SDS-PAGE; tertiary structure by intrinsic and extrinsic (ANS) fluorescence and zeta potential; secondary structure by ATR-FTIR; primary structure by mass spectrometry; and side-chain reactivity and protein-bound carbonyls by colorimetric assays. RP-HPLC profiles revealed treatment- and intensity-dependent losses in protein nativity in the order hydrogen peroxide (H2O2)/tert-butyl hydroperoxide (TBHP) > ultraviolet radiation (UVB) > elevated temperature (+40 °C). H2O2 and TBHP promoted oxygen addition at the primary-structure level, affecting secondary and tertiary structure. UVB mainly induced structural rearrangements, while elevated temperature caused partly hydrophobic modifications, increasing RP-HPLC retention times of modified proteins.
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