As the demand for dried timber has been increasing worldwide, drying of timbers is required to supply the demand in the manufacturing industries of furniture and other interior woodwork. During the drying process, some condensation will occur, resulting in liquids' emission, which requires disposal. In the liquid condensate, there are many substances at low concentrations that make their recovery uneconomic. After revealing the potential of the substances available in these condensates, biotechnological processes can be employed to produce high value-added products. This condensate contains tree sap, including proteins, enzymes, and many natural compounds such as terpenoids, alkaloids, phenolic compounds. Thus, it is considered that these natural compounds might be beneficial in pharmaceutical applications. Natural compounds such as phenolic compounds are bioactive substances that have antioxidant, antimicrobial, and cytotoxic activities. This study was aimed to characterize the condensate from the Rf-vacuum drying process of beech (Fagus sylvatica L.), walnut (Juglans regia L.), and restharrow (Onosis arvensis L.) in terms of antioxidant capacity, total phenol content, antimicrobial activity, and cytotoxicity. Obtained data revealed that condensates from the Rf-vacuum drying process inhibited the growth of both S. epidermidis and E. coli significantly.