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The aim of this study was to optimize extracion of flaxseed oil process by ultrasonic treatment and determination of its effect on ω-3 fatty acid. The extraction of flaxseed oil was optimized by using different solvents like methanol, acetone, petroleum ether, ethanol, hexane and dichloromethane. The ultrasonication treatment was optimized with respect to amplitude (20–80 kHz), temperature (25–40 °C), sonication time (20–80 min) and solid to solvent ratio (1:5, 1:10 and 1:15) for extraction of flaxseed oil. The extracted oil was subjected to GC analysis to determine ω-3 fatty acid. The recovery of flaxseed oil was higher with hexane followed by dichloromethane. The ultrasonic treatment at frequency of 40 kHz, temperature of 30 °C, extraction time of 40 min and solid to solvent ratio: 1:10 gave best results for extraction. The ultrasonic assisted extraction improves the extraction yield by 11.5% with similar amount of solvent. The chromatograph showed that there were no significant effects on α-Linolenic acid (ω-3) by the ultrasonic assisted extraction. This will beneficial to the oil extractor to recover higher amount of oil from same amount of the raw materials.