Purification and Characterization of Triacylglycerols in Natural Oils

Lipids, relatively nonpolar chemical substances found in plant, bacterial, and animal cells, are among the most ubiquitous of biomolecules. In this experiment, a lipid extract of ground nutmeg will be purified by chromatography on a silica gel column. Analysis of the lipid extract by thin-layer chromatography will provide the classification of the components in the extract. The unknown lipids will be further characterized by saponification and analysis of the fatty acid content by gas chromatography. For an abbreviated experiment, students may be provided samples of natural oils and fats that can be analyzed by saponification and gas chromatography. 

In this experiment we will approach the world of lipids with an introduction to analytical procedures currently applied to lipid extraction, purification, characterization, and structure elucidation. 

The extraction procedure used to isolate lipids from biological tissue depends on the class of lipid desired and the nature of the biological source (animal tissue, plant leaf, plant seed, bacteria, cell membranes, etc.). Because lipids are generally less polar than other cell constituents, they may be selectively extracted with the use of organic solvents. Early studies of lipids used ether, acetone, hexane, and other organic solvents for extraction however, these solvents extract only lipids bound in a nonpolar or hydrophobic manner. In the 1950s, Folch s group reported the use of chloroform and methanol (2 1) in 

Separation of crude lipid extracts into individual lipid classes is difficult and time-consuming. In some cases a crude separation of lipids can be attained by selective solvent extraction. For more extensive purification of lipids, the researcher must turn to chromatography. Chromatographic methods can both resolve a complex lipid mixture into the various classes of lipids and separate the individual components of a single class of lipids. 

Once several silica gel column fractions have been collected, the eluted fractions must be analyzed for the presence of lipids. Thin-layer chromatography on silica gel plates will be introduced in this experiment for the detection and identification of neutral lipids. 

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EXPERIMENT 6 Purification and Characterization of Triacylglycerols in Natural Oils 303... 

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