HPLC Analysis of Insulin (Species Variants) on Discovery^® BIO Wide Pore C8

column

Discovery BIO Wide Pore C8, 15 cm × 4.6 mm I.D., 5 μm particles (568322-U)

mobile phase

[A] (0.1% TFA in water):(0.1% TFA in acetonitrile), 71:29 [B]: (0.1% TFA in water):(0.1% TFA in acetonitrile), 68:32,

gradient

0-100% B in 30 min

flow rate

1 mL/min

column temp.

30 °C

detector

UV, 215 nm

injection

5 μL in 0.1% TFA containing 5ug each of three insulin variants