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. 2021 Aug 20;9:736788. doi: 10.3389/fchem.2021.736788

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Copyright © 2021 Geib, Moghaddam, Supinski, Golizeh and Sleno.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Reference samples (A) were prepared using non-treated mouse and rat liver homogenates and HP-IAM as the cysteine alkylation reagent. Modified peptides of interest from potential protein targets were selected for MRM method preparation. Final LC-sMRM methods were used to screen for modified peptides in (B) APAP-treated samples. Data-dependent high-resolution MS/MS experiments were performed on reference samples as well as selected APAP-treated rat and mouse samples for identifying modified peptides via database searching with custom modification (+C₈H₇NO₂ addition onto Cys residues).