Background: Zanthoxylum rhetsa (Roxb.) DC and Zanthoxylum limonella Alston are spices for flavouring in indigenous Thai food. They are traditionally used as an aromatic, astringent, antimicrobial, antiseptic and antidiabetic agent. The purpose of this study is to examine their chemical compositions and evaluate antibacterial, antioxidant and anticancer properties of the essential oils. Materials and Methods: The essential oils of Z. rhetsa and Z. limonella were analysed for phytochemical constituents by Gas chromatography-mass spectrometry (GC-MS). The antibacterial activity was determined against several bacteria using the microdilution method. Antioxidant capacity was determined by free radical scavenger 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) methods. The anticancer activity was determined with two breast cancer cell lines (MCF-7 and MDA-MB-231) and the normal African green monkey kidney epithelial (Vero) cell line and using MTT assay. Results: Sabinene (22.51%) and terpinene-4-ol (32.33%) were found to be major components of Z. rhetsa essential oil while limonene (57.94%) and alpha-phelladrene (15.54%) were the major components of Z. limonella essential oil. Essential oil from Z. limonella exhibited broad spectrum antibacterial activity. Z. rhetsa and Z. limonella essential oils exhibited moderate antioxidant activity. The essential oil from Z. rhetsa possessed the ability to inhibit breast cancer cell (MCF-7 and MDA-MB-231) proliferation and cell viability. Conclusion: This study suggest that the essential oils from Z. limonella and Z. rhetsa could be applied as safe antibacterial and antioxidant agents for food and have the potential for further development of new anticancer agents.