Detection of potential ion suppression for peptide analysis in nanoflow liquid chromatography/mass spectrometry

A detection technique for ion suppression in liquid chromatography/mass spectrometry (LC/MS) was developed by adding a probe to an LC mobile phase at a certain concentration. The probe is so hydrophilic that it is not adsorbed in a reversed-phase nanoflow LC column, and, furthermore, has an isoelectric point of about 3, which is lower than that for most peptides and is close to the pH of the mobile phase. The intensity of the protonated probe molecule decreases much more than that of other peptides when ion suppression occurs. Thus, the occurrence of the ion suppression is detected by a decrease in the mass chromatogram for the protonated probe molecule, and the decrease ratio is higher than that for other ions.