Apohemoglobin, carboxymethylated, tryptic digest

column

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

mobile phase

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

gradient

0-100% B in 65 min

flow rate

1 mL/min

column temp.

30 °C

detector

UV, 215 nm

injection

50 μL

sample

carboxymethylated apohemoglobin tryptic digest in 50 mM ammonium bicarbonate