BACKGROUND The presence of motor signs and symptoms in Parkinson's disease (PD) is the result of a long-lasting prodromal phase with an advancing neurodegenerative process. The identification of PD patients in an early phase is, however, crucial for developing disease-modifying drugs. The objective of our study is to investigate whether Diffusion Tensor Imaging (DTI) of the Substantia nigra (SN) analyzed by machine learning algorithms (ML) can be used to identify PD patients. METHODS Our study proposes the use of computer-aided algorithms and a highly reproducible approach (in contrast to manually SN segmentation) to increase the reliability and accuracy of DTI metrics used for classification. RESULTS The results of our study do not confirm the feasibility of the DTI approach, neither on a whole-brain level, ROI-labelled analyses, nor when focusing on the SN only. CONCLUSIONS Our study did not provide any evidence to support the hypothesis that DTI-based analysis, in particular of the SN, could be used to identify PD patients correctly.