Fractal analysis of images of the retinal vasculature is an instrument that has proven to be of great value both for the characterization of various pathologies and for the study of the vasculature in healthy retinas. To quantify this parameter, it is necessary to consider the treatment of the fractal object and the analysis conditions to ensure the validity of the results. Fractal and multifractal analysis of the retinal vasculature depends on several factors, including the fractal methods applied, the segmentation algorithm and calculation used, and especially the quality of the retinal image which directly influences the accuracy of the segmentation. These factors can influence the calculation and analysis of the fractal or multifractal dimensions. In the present work, different metrics have been developed to quantify the differences introduced by different segmentation methods and image datasets. Using the developed metrics, it has been possible to determine and quantify the influence of the factors studied effectively. The results indicate that the developed metrics allow to quantify these differences, as well as provide criteria on which are the best methods and protocols, which is relevant when using fractal and multifractal methods as an aid in retinal characterization and in the diagnosis of different anomalies.