In this paper, surface-enhanced Raman spectroscopy (SERS) was used to investigate the adsorption process of cysteine (Cys). Studies were carried out in the presence of phosphate-buffered saline solution (PBS), at pH 7.4, and acidified to pH 5, 3, and 1, on the surface of Ti for implant application. In situ SERS spectra obtained for the Cys/Ti solution system, after 24 h of immersion time, indicated that the buffer solution strongly influences the adsorption behavior of Cys on the Ti surface. This results in a decrease in Cys adsorption on the Ti surface, in the range of pH 7.4 to 3. The strong interaction between a sulfur atom of Cys and a Ti surface was observed only at pH = 1, under strongly acidic conditions. In contrast, ex situ SERS spectra recorded for the same samples but in a dried Cys/Ti system show a completely different behavior of Cys on the Ti surface. Formation of a disulfide (S-S) bond has occurred as a result of the dimerization or aggregation of Cys molecules on the Ti surface. Detailed analysis of the adsorption behavior of Cys on the Ti surface can be very important in the preparation of bioactive materials (i.e., coated by organic layers).