FSCN 4613 Lecture Notes - Lecture 4: Pasteur Pipette, Petroleum Ether, Lipoprotein
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Concentrate your lipids to make it easier to apply to the tlc plate- serum is mostly water! To remove contaminants that would interfere with the separation. Add salt solution to create two phases. Upper phase- methanol: water (1:1) contains water-soluble compounds. Lower phase- chloroform (contains lipids that we are interested in) Sinks to the bottom because it is very heavy. Remove upper phase, wash interface with methanol: water. Reconstitute lipids in petroleum ether, transfer to vial for storage. Use a mixture of organic solvents- chloroform and methanol. Filtrate will be collected (which contains lipids) Add sodium chloride solution to form two phases. Upper phase - methanol & water - contains sugar, amino acids, other water-soluble molecules. These are the things we don"t want. Remove the upper phase with a pasteur pipette. Add clean upper phase, then remove it to wash interface of two phases.