I am extracting glycosaminoglycan from fish viscera of Seabass and I am facing some difficulties, as I kept tissue in acetone for more than one month but still I am getting lots of fat and not proper precipitation of GAG is taking place. Please help me out with some suggestions.
Dear Ravi,
acetone is not good solvent of fats, but it can sufficiently dewater your sample prior to extraction of fats with non polar solvents.
You should use less polar solvent to remove fats completely from the dewatered sample, e.g. hexane, CHCl3 or maybe THF. After storing the samples in any of these solvents, you may wasch out the solvents from the sample with acetone and then start extracion of GAG, followed with its precipitation.
Note: to remove the fats completely, use at least 3 or multistep exractions of the sample prior to GAG isolation. If you can use soxhlet extractor, maybe 8 to 16 h of continuous extraction might do.
Good luck.
Ravi,
as for acetone used for deflation, you need much time and many extraction steps to remove most of fat present in the fish sample.
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