HPLC Analysis of Steroids on Ascentis^® Express C18 with Methanolic Mobile Phase

column

Ascentis Express C18, 10 cm x 2.1 mm I.D., 2.7 μm particles (53823-U)

mobile phase

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

flow rate

0.2 mL/min

column temp.

35 °C

detector

UV, 254 nm

injection

2 μL

sample

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