HPLC Analysis of Prednisolone (Secondary Pharma Standard) on Ascentis^® C18

column

Ascentis C18, 15 cm x 4.6 mm I.D., 5 μm particles (581324-U)

mobile phase

[A] water; [B] methanol:acetonitrile (50:50, A:B)

gradient

40% B for 14 min; to 80% B in 6 min; held at 80% B for 5 min

flow rate

1 mL/min

column temp.

40 °C

detector

UV, 254 nm

injection

10 μL

sample

125 μg/mL in water:acetonitrile (60:40)