Here we use a quantitative label-free proteomics workflow - "one-pot" enrichment - for the comprehensive identification and quantification of lysine acetylated and succinylated peptides from a single sample. Coupled with label-free, data-independent acquisition (DIA), we identified and quantified 2235 acetylated and 2173 succinylated peptides from mouse liver protein. We show peak areas are highly correlated between one-pot and single-antibody enrichments. Finally, we show this method enables the detection of acetylation and succinylation on the same peptide.