HPLC Analysis of Insulin (Species Variants) 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]: 71:29, (0.1% TFA in water):(0.1% TFA in acetonitrile);

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

gradient

0-100% B in 30 min

flow rate

1 mL/min

column temp.

ambient

detector

UV, 215 nm

injection

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