Since, I do not have standard steroid (Androstanedione), I was wondering that if any one can tell me what is the TLC mobile phase for androstanedione and is it UV-active or not. I want to check androstanedione as product in my assay mixture using TLC and HPLC method.
TLC was carried out with Kieselgel 60 F254 plates (Merck, Darmstadt, Germany) using hexane/acetone (3:1 v:v) . In order to develop the image, the plates were sprayed with solution of methanol in concentrated sulfuric acid (1:1) and heated to 120 °C for 3 min.
It may be UV active and give blue fluroscence spot. As it contain lactone ring and most of the steroidal compund consisiting lactone ring is UV active.
You can use hexane or methanol as TLC mobile phase for androstanedione. shows photomeric effect and do not change in UV-light, but in visible light become inactive. You can check other solvents for finding new solution..
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