Piperazine exists in a number of energetically close structural conformations, and here, we investigated the dependence of their relative abundance on the surrounding conditions by using Raman and SERS  spectroscopy in pure solid, aqueous solution and Ag hydrosol. The experimental results were interpreted by DFT calculations using B3LYP functional with aug-cc-pvdz/LANL2DZ basis sets. In the chair form of piperazine, which is more stable than the skewed boat by ~8 kcal mol^-1, the two N–H bonds can remain equatorial or axial, leading to three different conformations, eq–eq, eq–ax and ax–ax. The calculated Raman spectrum of the lowest energy eq–eq conformation corresponds well with the experimental spectrum in pure solid, indicating eq–eq to be predominant. But, the contribution of the eq–ax conformation was found to be maximum in aqueous solution. The SERS spectrum revealed that eq–ax conformation was preferably adopted as piperazine was adsorbed vertically through its axial N-atom over silver nanoparticle surface.