Piper betle leaf oil shows cell cytotoxicity and antioxidant activity in vitro

Piper betle leaf essential oil showed pronounced free-radical-scavenging activity in four laboratory assays and concentration-dependent cytotoxicity against human oral carcinoma cells in an in vitro study. In the MTT cell-viability test, the researchers evaluated oil concentrations from 6.25 to 100 µg/mL and determined an IC50 of 22.8 µg/mL for KB cells. The study also assessed membrane damage, adding a second measure alongside the observed loss of cancer-cell viability.

The researchers examined antioxidant activity using DPPH, hydrogen peroxide, superoxide dismutase and ABTS assays. The essential oil produced pronounced scavenging activity across all four tests, according to the study summary. Because the available report does not provide assay-specific numerical results, it does not allow direct comparison of the magnitude of activity among the four methods or calculation of an overall effect size.

The summary also does not report the detailed chemical composition of the tested oil, limiting attribution of the measured effects to particular constituents. The cancer-cell experiment used KB, a human oral carcinoma cell line. MTT testing covered concentrations ranging from 6.25 to 100 µg/mL, a 16-fold span between the lowest and highest doses.

The reported IC50 was 22.8 µg/mL, while viability declined as the oil concentration increased. The investigators described this pattern as significant, concentration-dependent cytotoxicity and considerable antiproliferative activity. Their separate membrane-damage assessment also indicated an effect on KB cells, although the supplied summary does not identify the procedure or provide quantitative membrane results.

The authors proposed that the oil’s many bioactive components, including functional groups associated with free-radical scavenging and inhibition of cancer cells, may account for the observed activity. The results therefore support further investigation of Piper betle leaf essential oil as a material with laboratory-measured antioxidant and cytotoxic activity, not a conclusion that it acts as a therapeutic anticancer agent. This was a laboratory study conducted in vitro, with testing in chemical assay systems and one cultured human cancer cell line.

It did not evaluate treatment of cancer in animals or humans, and the supplied evidence does not establish effects on normal cells, selectivity between cancer and non-cancerous cells, absorption, dosing, safety, efficacy or clinical benefit. No basis is provided for recommending ingestion, inhalation or topical use of the essential oil. Taken together, the findings offer a preliminary experimental signal across four free-radical-scavenging assays and KB-cell viability and membrane assessments, while questions about composition, mechanism, selectivity and translation to living organisms remain unresolved.

Subject of Research: In vitro antioxidant and cytotoxic testing of Piper betle leaf essential oil

Article Title: In vitro assessment of the free radical scavenging and cytotoxic activities of Piper betle leaf essential oil

Article References: Janani, P., Anju, A., Anjana, A., Babu, R. D., & Meiyalagan, V. (2026). In vitro assessment of the free radical scavenging and cytotoxic activities of Piper betle leaf essential oil. Medical Oncology, 43(11), Article 336. https://doi.org/10.1007/s12032-026-03462-9

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DOI: 10.1007/s12032-026-03462-9

Keywords: Piper betle, leaf essential oil, free-radical scavenging, antioxidant assays, oral carcinoma, KB cells, cell viability, cytotoxicity, MTT assay, in vitro research, vitro, assessment