Table 1. Optimization of HILIC LC–MS Separation of PSA Peptides and Glycopeptides^a.

protease	level	% AcN (50–95%)	pH (3.6–5.2)	[AF] (5–50 mM)
trypsin	peptides	increased retention with increased % AcN, minimum 90%	higher pH gives better peptide separation	no effects on retention, lower signals intensity with higher AF concentration
	glycopeptides	increased retention with increased % AcN, minimum 80%	higher pH gives less retention, but slightly better signal intensity	no effects on retention, lower signals intensity with higher AF concentration
	optimal	90% can	4.4	10 mM
ArgC	peptides	increased retention with increased % AcN, minimum 90%	lower pH gives better peptide separation	variable effects on retention, lower signal intensity with higher AF concentration
	glycopeptides	increased retention with increased % AcN, minimum 80%	higher pH gives less retention, but slightly better signal intensity	no effects on retention, lower signals intensity with higher AF concentration
	optimal	90% can	4.2	10 mM

^aEffects of changes in solvent composition are indicated together with the eventual optimal solvent conditions.