In the current study, Raman spectroscopy is employed for the identification of the biochemical changes taking place during the development of Hepatitis C. The Raman spectral data acquired from the human blood plasma samples of infected and healthy individuals is analysed by Principal Components Analysis and the Raman spectral markers of the Hepatitis C Virus (HCV) infection are identified. Spectral changes include those associated with nucleic acidsat720 cm−1, 1077 cm−1 1678 (CO stretching mode of dGTP of RNA), 1778 cm−1 (RNA), with proteins at 1641 cm−1(amide-I), 1721 cm−1(CC stretching of proteins) and lipids at 1738 cm−1(CO of ester group in lipids). These differences in Raman spectral features of blood plasma samples of the patients and healthy volunteers can be associated with the development of the biochemical changes during HCV infection