Brittle stars, unlike most other echinoderms, do not use their small tube feet for locomotion but instead use their flexible arms to produce a rowing or reverse rowing movement. They are among the fastest-moving echinoderms with the ability of complex locomotory behaviors. Considering the high species diversity and variability in morphotypes, a proper understanding of intra- and interspecies variation in arm flexibility and movement is lacking. This study focuses on the exploration of the methods to investigate the variability in brittle star locomotion and individual arm use. We performed a two-dimensional (2D) image processing on horizontal movement only. The result indicated that sinuosity, disc displacement and arm angle are important parameters to interpret ophiuroid locomotion. A dedicated Python script to calculate the studied movement parameters and visualize the results applicable to all 5-armed brittle stars was developed. These results can serve as the basis for further research in robotics inspired by brittle star locomotion.