Extration and Purification of Total Lipids from Animal Tissues.docVIP

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Extration and Purification of Total Lipids from Animal Tissues.doc

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REWIEW: Extraction and Purification of Total Lipids from Animal Tissues 1.INTRODUCTION Lipid research is an old branch of organic chemistry.It was only in the 1950s with the work of Folch and coworkers , Sperry and Brand , and Bligh and Dyer that satisfactory extraction methods were developed. The existing methods were considered but none was entirely satisfactory. The methods of Folch et al. were too time-consuming for routine investigations and since the former method entailed heating and evaporation it was considered unsuitable for lipid composition studies. The modified method of Folch et al. was more rapid than their previous method but still had the disadvantage of employing large and inconvenient volumes of solvent. The method used by Dyer and Morton was rapid but extracted only a fraction of the total lipid.These methods, while extracting adequately the lipids from the tissues, did not yield an extract entirely free of non-lipid. It was only in the middle 1960s with the introduction of the dextran gel purification techniques developed by Wells and Dittmer , Siakotos and Rouser , and Wuthier , that it was possible to have confidence in both the extraction and purification of lipids from a wide variety of tissues and organisms. There summary is to describe the of the methods for lipid extraction and purification and their advantages and disadvantages. 2.THE EXTRACTION OF TISSUE LIPIDS 2.1 REAGENTS Chloroform. Reagent grade . Methanol. Reagent grade. For use with tissues relatively poor in lipides, such as muscle or blood plasma, both the chloroform and methanol must be redistilled. Chloroform-methanol mixture. 2:1 by volume. Pure solvents upper phase and pure solvents louer phase. Chloroform , methanol, and water arc mixed in a separatory funn el in the proportions 8 :4:3 by volume. When the mixture is allowed to stand, a biphasic system is obtained. The two phases are collected separately and stored in glass bottles. It has been found that the approx