HPLC Analysis of Carboxymethylated Apohemoglobin Tryptic Digest on Discovery^® BIO Wide Pore C5

column

Discovery BIO Wide Pore C5, 15 cm × 4.6 mm I.D., 5 μm particles (568422-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