In order to promote the development and utilization of the liver， a by-product of Takifugu rubripes' processing， high content of n-3 polyunsaturated fatty acid fish oil was obtained from Takifugu rubripes' liver by enzymatic method. The sensory properties， basic physicochemical properties， tetrodotoxin content， fatty acid composition， liposoluble substances， and volatile compounds of fish oil before and after refining were analyzed. The results showed that the crude and refined fish oil meet the standards of grade II crude fish oil and grade I refined fish oil in the aquatic industry standard respectively； No tetrodotoxin was detected in both crude and refined fish oil， and 20 kinds of fatty acids were detected in both， with unsaturated fatty acids as high as 71.50% and 71.11%， the total contents of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were 25.98% and 25.64%， respectively； The total amount of tocopherol in refined fish oil decreased slightly， but it could still reach 1.51 mg/100 g. 38 volatile compounds were detected in both oils by gas chromatography ion mobility spectroscopy (GC-IMS) system， including 8 alcohols， 1 ether， 18 aldehydes， 8 ketones， and 3 esters； 31 different volatile compounds (VIP>1) were screened by orthogonal partial least squares discriminant analysis (OPLS-DA) combined with variable importance in the projection (VIP)； Quantitative descriptive analysis (QDA)， differential spectrogram and thermographic cluster analysis all showed that compared with crude fish oil， aldehydes， ketones and other undesirable flavor substances in refined fish oil were significantly reduced. This study provides a theoretical basis for Takifugu rubripes liver oil as a new type of fish oil.