Although blossom and honeydew honeys are produced from different nectar sources, it is very difficult to distinguish them. In this study, physicochemical and biochemical properties were compared to distinguish honeydew and blossom honeys obtained from different botanical sources. The honeys of pine (Pinus spp.) and oak (Quercus, spp.) were used as honeydew honey, and chestnut (Castania sativa L.) and highland honey were used as blossom honey. Turbidity, specific optical rotation (SOR) [α]20, electrical conductivity (EC), CIEL* a*b color parameters, and moisture and pH were assessed as physical parameters. Proline, invertase activity, sugar composition, total phenolic and flavonoid contents, and antioxidant capacity were used as biochemical properties. It was determined that SOR values were the most important parameters in distinguishing honeydew and blossom honeys. Turbidity stands out as a crucial factor in differentiating between pine and oak honeys. Except for chestnut honey, electrical conductivity, CIE color Lab values, and sugar profiles were identified as additional distinguishing factors among the tested honeys.