Bcakground: Extracts obtained from natural sources such as plants are of immense importance for humans. Methods: Therefore this study was conducted to obtain essential oil from the seeds of T. ammi by conventional and non-conventional methods. Hydrodistillation (HD), Solvent Extraction (SE), Ultrasonication (US), and Supercritical Carbon-dioxide (SC-CO₂) extraction techniques were used to extract essential oil from the powdered seeds of T. ammi. A quality control method for each extracted oil was developed using HPTLC, FTIR, and GC-MS. The optimization process was carried out using fractional factorial design (FFD) under which three parameters were considered: pressure (150, 175, and 300 bar), temperature (25, 30, and 40 °C), and CO₂ flow rate (5, 10, 15 g/min). Results: The yield of essential oil obtained from the HD, SE, US, and SC-CO₂ methods were 1.20%, 1.82%, 2.30%, and 2.64% v/w, respectively. Antioxidant activity was determined by the DPPH and superoxide scavenging methods and the IC50 (Inhibition Concentration) values of the T. ammi oil sample were found to be 36.41 and 20.55 µg mL-1, respectively. Conclusion: The present paper reported that different extraction methods lead to different yields of essential oils and the choice of a suitable method is extremely important to obtain more preferred compounds. The yield was higher in the SC-CO₂ method and it is a sustainable and green extraction technique. Many important constituents were detected in analytical techniques. Antioxidant activities carried out showed that essential oil extracted from T. ammi seeds possess significant antioxidant activity.
Keywords: FTIR; GC-MS; HPTLC; SC-CO2; optimization; sustainable.