Peptides

column

Discovery BIO Wide Pore C18, 15 cm × 4.6 mm I.D., 5 μm particles (568222-U)

mobile phase

[A] (95:5), 0.1% TFA in water:0.1% TFA in acetonitrile; [B] (50:50), 0.1%TFA in water:0.1% TFA in acetonitrile

gradient

0-100% B in 65 min

flow rate

1 mL/min

column temp.

30 °C

detector

UV, 215 nm

injection

50μL carboxymethylated apohemoglobin tryptic digest in 50 mM ammonium bicarbonate