Ruta chalepensis L. is plant characteristic for the Mediterranean region but is widely distributed and cultivated worldwide, especially in tropical areas. The essential oil of R. chalepensis L. is often used for medicinal purposes due to its aromatherapy and pharmaceutical properties. Despite its numerous benefits, R. chalepensis contains some toxic compounds, necessitating careful handling and usage. Essential oils are mixtures of volatile compounds with distinct and characteristic scents. They are obtained from various parts of the plant and are known for their aromas and medicinal properties. Gas chromatography hyphenated with mass spectrometry is one of the most used techniques for analyzing volatile compounds in essential oils. This technique allows for very precise qualitative and quantitative analysis, even when only a small amount of the sample is available. This thesis describes the isolation and analysis of the essential oil of R. chalepensis L. grown on a cell line. The essential oil was isolated from plant material using water distillation, and analysis was carried out using gas chromatography-mass spectrometry. In all three essential oils, the most prevalent compounds are undecan-2-one, tridecan-2,4-dione, 2- undecyl acetate, and decanal. Furanocoumarins with toxic properties, including 3-(α,α-dimethylallyl)- psoralen (kalepensin), psoralen, methoxsalen, and bergapten, were also identified.