The application of human bone in forensic proteomics is an expanding novel purpose in the aim of successfully quantifying biological estimations used in medico-legal investigations with greater accuracy. In this project, different extraction protocols were tested on n=30 human bone samples, including S-Trap technology with two different lysis buffer, and ZipTips. Additionally, a comparison was made between data-dependent acquisition (DDA) and data-independent acquisition (DIA) mode. Results showed less missing data using S-Traps instead of the more routine reverse-phase media tips (ZipTip). The type of lysis buffer when using S-Traps does not impact largely the analysis conducted in forensic proteomic workflows. Lastly, it was found that when open-source software is used for data processing for both DDA and DIA modes, DIA has the upper advantage in terms of acquiring a larger number of proteins due to its greater sensitivity to those with a lower abundance.