HPLC Analysis of Steroids on Ascentis^® Express F5

Ascentis Express F5, 15 cm x 4.6 mm I.D., 2.7 μm particles (53591-U)

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

0.8 mL/min

2393 psi (165 bar)

35 °C

UV, 240 nm

5 μL

10 μg/mL of each in 90: 10, water: methanol

Sample concentrations at 10 ug/mL in 90:10 water:methanol. Acetonitrile does not provide adequate selectivity on this phase. Mass spec was used for peak identification.

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany