Multiplexed Data-Independent Acquisition (plexDIA) aims to bridge the gap between the quantitative depth of DIA and the sample throughput of isobaric tag-based workflows. Here, we implement non‑isobaric sample multiplexing using commercially available, isotopically differentiated tandem mass tag (TMT) analogues (TMTpro Zero, standard, and super heavy) to perform plexDIA. By introducing defined mass offsets, we avoid reporter ion interference and we shift signal deconvolution to the MS1 and precursor fragment levels. We benchmarked this workflow using the yeast triple knockout (TKO) standard (Δmet6, Δpfk2, Δura2) analyzed on an Astral mass spectrometer. Notably, targeted quantification closely mirrored biological ground truth, with protein knockouts exhibiting near‑zero intensities strictly within their assigned labeling channels and no detectable cross‑channel signal bleeding. Ultimately, non-isobaric plexDIA avoids the pitfalls of isobaric ratio compression, and provides a highly accurate and scalable framework for quantitative proteomics.