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Aims: Oncology treatments aim at selective toxicity for tumor (compared to normal) cells, and chromone- coumarin hybrids have shown such activity. Methods: In this study, we test a novel series of synthetic chromone and coumarin derivatives (1–9) for cytotoxic activity against a panel of tumor cell lines (MCF-7, A549, HepG2, HTC-116, B16 and Caco-2) opposed to non-tumor cells (HEK-293t). Electrical impedance spectroscopy was used to monitor cell viability in real time. Results: Compound 8 showed the most potent activity, and it significantly diminished cancer cell proliferation and viability in different cell lines. It induced apoptosis in a dose-dependent manner, as shown by Western blot and flow cytometry. Conlcusion: Electrical impedance spectroscopy appears to be a convenient tool for in vitro cytotoxicity analysis, which could be useful for identifying drug effects and side effects during early phases of drug discovery and development.