In order to estimate the Fatty acids content of cooked fish, Bligh and Dyer extraction was used to extract fat. The homogenate samlpe chloroform methanol and water mixture gives a cream white homogenate which separates when centrifuged. Fat extract gives very little total fat and I doubt whether the fat has not been properly derived into the chloroform layer since creaming happened while homogenizing.
Can anyone explain and suggest a solution
Dear Thiruchenduran,
you may wish to try the method developed by Smedes (in 1999). It avoids the use of (toxic) chlorinated solvents according to the Montreal Protocol (1987). Lipids are extracted from marine tissues using a propan-2-ol - cyclohexane - water (8:10:11, v/v/v) mixture followed by centrifugation. Due to its lower density, cyclohexane separates on top of the extraction mixture and can easily be removed. We applied the method successfully with high recoveries as regards lipids and associated POPs (dioxins, PCBs). Please refer to: Smedes F: Determination of total lipid using non-chlorinated solvents. Analyst 124 (1999) 1711–1718. You could also ask the author for a copy here: Article Determination of total lipid using non-chlorinated solvents
Wishing you best of success,
Johannes
Thank you for your reply. Dr. Johannes Haedrich . I have requested for the method. We will also has to run a reference sample using this method for adotion at our laboratory. I really appreciate your reply. Thanks again
Thiru
You are welcome. Did you already receive the method? It is especially designed for lipid extration from fish.
I have already requested but yet to receive the method.