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1. Essential Oils from Côa Valley Lamiaceae Species: Cytotoxicity and Antiproliferative Effect on Glioblastoma Cells

2. Since ancient times and across all populations, the therapeutic value of plants was validated by their wide use in traditional medicine to treat various diseases, including cancer. The well-known therapeutic significance attributed to plants usually resides in the secondary metabolites they produce Essential oils (EOs), obtained from aromatic plant species, are examples of complex mixtures of volatile secondary metabolites with huge therapeutic potential . They are produced and stored in specialized structures, internal or external, and are characterized by the presence of small molecular weight oxygenated compounds and hydrocarbons, mainly monoterpenes and sesquiterpenes . These compounds are well-known for their therapeutic value, including antitumor activit

3. Lavandula pedunculata (Mill.) Cav., Mentha cervina L. and Thymus mastichina (L.) L. subsp. mastichina are representative species of the Côa Valley’s flora, a Portuguese UNESCO World Heritage Site. L. pedunculata and T. mastichina are traditionally used to preserve olives and to aromatize bonfires while M. cervina is mainly used as a spice for river fish dishes. Despite their traditional uses, these aromatic plants are still undervalued, and literature regarding their bioactivity, especially anticancer, is scarce

4. Essential oils (EOs) from Lavandula pedunculata and Mentha cervina induce apoptosis in a dose-dependent manner, showing strong cytotoxic and antiproliferative potential against tumor cells, particularly in glioblastoma (GB) cells

5. Mentha cervina essential oil exhibited a slightly lower cytotoxic effect on tumor cells compared to Lavandula pedunculata and did not affect the viability of non-tumor cells Thymus mastichina essential oil did not induce a strong cytotoxic effect on GB cells The study used scanning electron microscopy (SEM) to assess the morphology of secretory structures and gas chromatography-mass spectrometry (GC-MS) to characterize the composition of essential oils

6. Retention time in Gas Chromatography-Mass Spectrometry (GC-MS) refers to the time taken for a compound to elute out of the chromatographic column. The retention time of a compound is used to identify and characterize substances within a sample In GC-MS analysis, the retention time reflects the interaction between the compound and the stationary phase of the column. Compounds with different chemical properties will have varying retention times

7. The retention index is calculated based on the retention times obtained for a mixture of n-alkanes (C8-C40) analyzed under identical conditions By comparing the retention times of the compounds in a sample to known standards, scientists can determine the identity of the substances present in the essential oils from Côa Valley Lamiaceae species. Retention time data are crucial for the accurate identification and quantification of compounds present in essential oils, aiding researchers in understanding the chemical composition and potential bioactivity of the samples