Steroids represent a broad class of natural products playing crucial roles in different biological systems and steroid derivatives were extensively used in the development of therapeutic agents. Steroid androsterone (ADT) derivatives, especially those substituted at position 3, were found to inhibit 17β-hydroxysteroid dehydrogenase type 3, a key enzyme involved in the formation of androgen testosterone from the natural substrate 4-androstene-3,17-dione. The enzyme inhibitory activity of ADT derivatives is however greatly modulated by the orientation (stereochemistry) of the substituent at position 3, it is consequently very important to determine the stereochemistry by using a powerful methodology such as nuclear magnetic resonance (NMR) spectroscopy. We report the ¹³C NMR data of thirty eight (38) ADT and epi-ADT derivatives, as alcohol or ether derivatives, bearing a substituent (alkyl, vinyl, allyl, phenyl, alkylphenyl, hydroxypropyl, bromopropyl or lactone) at position 3β or/and 3α. The carbon assignments were done using a combination of 1D and 2D NMR experiments, thus providing interesting markers for both the stereochemistry and nature of the substituent at position 3. Only the chemical shifts of carbon atoms (C1, C2, C4, C5, C10, and C19) surrounding position 3, as well as C3 itself were affected by the presence of a substituent. The magnitude of the deshielding effect observed for C3 was important and is helpful in the identification of a given stereoisomer, which information is not accurately predicted when using commercially available software.