HPLC Analysis of Steroids on Ascentis® Express F5
CONDITIONS
column
Ascentis Express F5, 15 cm x 4.6 mm I.D., 2.7 μm particles (53591-U)
mobile phase
[A] water; [B] methanol; (50:50, A:B)
flow rate
0.8 mL/min
pressure
2393 psi (165 bar)
column temp.
35 °C
detector
UV, 240 nm
injection
5 μL
sample
10 μg/mL of each in 90: 10, water: methanol
Description
Analysis Note
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.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany