HPLC Analysis of Peptides on Ascentis^® RP-Amide

column

Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 μm particles (565324-U)

mobile phase

[A] 0.1% trifluoroacetic acid in water; [B] 0.1% trifluoroacetic acid in acetonitrile

gradient

9 to 21% B in 5 min; to 27% B in 15 min

flow rate

1.0 mL/min

column temp.

30 °C

detector

UV, 220 nm

injection

5 μL

sample

as indicated in mobile phase A