HPLC Analysis of Steroids on Ascentis^® Express F5 with Methanolic Mobile Phase

column

Ascentis Express F5, 10 cm x 2.1 mm I.D., 2.7 μm particles (53569-U)

mobile phase

[A] water [B] methanol; (40:60, A:B)

flow rate

0.2 mL/min

column temp.

35 °C

detector

UV, 254 nm

injection

0.2 mL/min

sample

50 mg/L in 75:25, water:methanol