Apohemoglobin (horse), carboxymethylated, tryptic digest

column

Discovery BIO Wide Pore C18, 15 cm × 2.1 mm I.D., 3 μm particles (568222-U)

mobile phase

[A] 0.1% TFA in water; [B] 0.1% TFA in water:0.1% TFA in acetonitrile, 50:50;

gradient

0-100% B in 50 min

flow rate

0.2 mL/min

column temp.

35°C

detector

UV, 220 nm

injection

5 μL, each at 0.8 mg/mL in 0.1% TFA