Re-searching Dataset 3 with both off-by-one error anticipation, and sodium adduction enabled in GlyCombo reduced the number of unassigned MS2 scans by 40% compared to a protonated adduct search (Fig ). Although this complicated search comes at a cost of a 4x greater search time, the total search time was a fraction of the 200-minute-long LC-MS run. As the off-by-one search only yielded an 11% decrease in unassigned MS2 scans, this option could be skipped for data generated by instruments with accurate monoisotopic peak detection . A limitation of these more complex searches is the increase in false positives due to mistaking protonated adducts for sodiated adducts and correct precursor masses for off-by-one errors. As GlyCombo is the initial step in filtering candidate precursor ions for quantitation and characterization, users can opt for a higher false positive rate to ensure a high true positive rate. Therefore, it can be crucial to detect a comprehensive range of glycans to leverage subsequent quality assessments, including retention time filtering (Skyline), MS2 scoring (GlycoWorkBench), and precursor isotopic distribution evaluation (Skyline).