HPLC Analysis of Cytochrome c Species Variants on Discovery^® BIO Wide Pore C18

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); (70:30), [B] (0.1% TFA in water):(0.1% TFA in acetonitrile), 64:36

gradient

0-100% B in 30 min

flow rate

1 mL/min

column temp.

ambient

detector

UV, 220 nm

injection

12 μL each at 0.8 mg/mL in 0.1%TFA