HPLC Analysis of Calphostin C on Discovery^® HS C18

column

Discovery HS C18, 25 cm × 2.1 mm I.D., 5 μm particles (568503-U)

mobile phase

[A] (0.1% phosphorus acid in water):(0.1% phosphorus acid in acetonitrile); [B] 0.1% phosphorus acid in acetonitrile; (50:50, A:B)

flow rate

0.2 mL/min

column temp.

24 °C

detector

UV, 254 nm

injection

3 μL

sample

Calphostin C (0.25 g/L) in 40% MeOH, 0.025% TFA