In the present investigation, the hooded cuttlefish Sepia prashadi were collected from Mudasalodai landing centre, east coast of Tamil Nadu, India. Collected animals were dissected for their liver (digestive gland) and the dried liver was ground well and the cuttlefish liver oil was extracted. The percentage of oil yield was noted as 32.45 %. The extracted liver oil was used for estimating total lipids, triglycerides, total cholesterol, High Density Lipoproteins cholesterol (HDL), Low Density Lipoproteins cholesterol (LDL), Very Low Density Lipoproteins cholesterol (VLDL) and free fatty acids. The fatty acid composition of the liver oil such as Saturated Fatty Acids (SFA), Mono Unsaturated Fatty acids (MUFA) and Poly Unsaturated Fatty acids (PUFA) was analyzed using GC-MIS. The results of the present study revealed that the cuttlefish (S. prashadi) liver oil was found to contain more MUFA and also contain considerable amount of triglycerides and cholesterol. The cuttlefish oil can be used as a bio enrichment of live feeds and the residual waste during oil production can be used as manure, poultry feed, bio diesel production, etc. Moreover production of oil from the animal wastes such as liver is an eco-friendly process and can reduce the environment pollution. The quality of oil can be improved to increase the PUFA content for human consumption. Apart from all the above advantages, cuttlefish oil can be used as an alternative to fish oil, as it is very easy and cheap to produce than the fish oils.