In this study, sulphated polysaccharides were investigated in respect to their blood compatibility properties (hemocompatibility). Pure chitosan was treated with sulphating agents such as SO(3)/pyridine complex and chlorosulfonic acid (HClSO(3)) to obtain 3,6-O-sulfochitosan with low and high concentration of sulfur. These synthetically derived materials and the commercially available sulphated polysaccharides heparin and dextran sulfate, both with high concentrations of sulfur, were coated onto PET foils to act as surfaces with strong antithrombotic activity. This treatment should lead to better blood compatibility properties of PET materials for medical applications. To examine this, the optimized free hemoglobin method was applied to determine the antithrombotic activity of these surfaces. Glass as the standard thrombotic surface and a heparin-coated PET surface as a surface well-known for its strong antithrombotic activity were used as internal references. The experiments showed that dextran sulfate and sulphated chitosan with high concentrations of sulfur demonstrated the same antithrombotic activity as heparin over the whole period of measurement time. In addition, a relationship between the sulfur concentration in these sulphated polysaccharides and their blood compatibility properties can be demonstrated in this article.