Analysis of Intermediates of Steroid Transformations in Resting Cells by Thin-Layer Chromatography (TLC)

Thin-layer chromatography (TLC) is a useful and convenient method for the analysis of steroids due to: its simple sample preparation, low time-consuming process, high sensitivity, low equipment investment and capacity to work on many samples simultaneously. Here we describe a TLC easy protocol very useful to analyze steroid molecules derived from a biotransformation carried out in wild-type and mutant resting cells of Rhodococcus ruber strain Chol-4. Following this protocol, we were able to detect the presence or the absence of some well-known intermediates of cholesterol catabolism in Rhodococcus, namely AD, ADD, and 9OHAD.