Biological studies on Mesolithic human remains from the Polish region are a rare subject of scientific research due to the limited number of these relics and their poor state of preservation. From the project titled “Old material with new methods: Using the latest bio-chemical analysis in studies of Mesolithic human remains from the Polish areas,” the radiocarbon (14C) dating of bones using accelerator mass spectrometry (AMS) has been performed. For these experiments, the gelatin was extracted from bones, and its quality evaluated by the C/Nat ratio and the stable isotope composition of both carbon and nitrogen. The 14C results have been obtained for 11 bone samples from 5 sites, and throughout this work the results of two preparation methods are compared. The simple gelatin extraction provided material with unsatisfactory collagen quality indicators, while additional alkali treatment allowed us to obtain much more reliable, and generally older, results. Additionally, analysis on VIRI/SIRI samples were conducted to test the developed method. Only seven of the investigated bone samples yielded ages within Mesolithic period, and the most reliable dates range from 5800 to 6800 cal BC. One sample was not datable, and two were shown to be much younger than expected.