Table 2.

Cleaning and desalting of peptides.

Peptide cleaning/desalting using C18 SepPac			(CV = 150 µL)
Initialize	3CV	1 x 450 µL	MeOH
Wash TFA	3CV	1 x 450 µL	50% ACN/0.1% TFA
Equilibrate	6CV	1 x 900 µL	0.1% TFA
Sample load (pH 1-3)	≥1CV	≥1 x 150 µL	in 0.1% TFA
Wash/desalt 1	3CV	1 x 450 µL	0.1% TFA/2% ACN
Wash/desalt 2	3CV	1 x 450 µL	0.1% FA/ 2% ACN
Elution 1	3CV	1 x 150 µL	30% ACN/0.1% FA
Elution 2	2CV	1 x 300 µL	80% ACN/0.1% FA

CV, Column volumes; MeOH, Methanol; TFA, Trifluoroacetic Acid; ACN, Acetonitrile; FA, Formic Acid.