While there are many ways to identify substructures in galaxy clusters using different wavelengths, each technique has its own caveat. In this paper, we conduct a detailed substructure search and dynamical state characterisation of Abell 2399, a galaxy cluster in the local Universe ($z sim 0.0579$), by performing a multi-wavelength analysis and testing the results through hydro-dynamical simulations. In particular, we apply a Gaussian Mixture Model to the spectroscopic data from SDSS, WINGS, and Omega WINGS Surveys to identify substructures. We further use public textit{XMM-Newton} data to investigate the intracluster medium (ICM) thermal properties, creating temperature, metallicity, entropy, and pressure maps. Finally, we run hydro-dynamical simulations to constrain the merger stage of this system. The ICM is very asymmetrical and has regions of temperature and pressure enhancement that evidence a recent merging process. The optical substructure analysis retrieves the two main X-ray concentrations. The temperature, entropy, and pressure are smaller in the secondary clump than in the main clump. On the other hand, its metallicity is considerably higher. This result can be explained by the scenario found by the hydro-dynamical simulations where the secondary clump passed very near to the centre of the main cluster possibly causing the galaxies of that region to release more metals through the increase of ram-pressure stripping.16