A pilot study on 22 samples corresponding to 220 pure standards from the MCEBIO compound library was conducted to evaluate the influence of the AGC target and mass resolution, focusing on MS level ≥ 3. Accordingly, the settings for MS 1 and MS 2 remained unchanged whereas the other MS levels were affected. In Method 1, the AGC target was set to 60% (30k), which is higher than the recommendation for small molecules by the instrument's manufacturer, specifically MS3-OT AGC for small molecules between 20% and 40% (10-20k).