Lipid
Analysis of Lipid through Two-Dimensional Thin Layer Chromatography
Myca Pua, Ramon Ramos, Amanda Redilas, Kimleigh Reyes,
Nathaniel Sim and Clara Tamondong
Group 9 2F Medical Technology Biochemistry Laboratory
ABSTRACT
Lipids constitute a group of naturally occurring molecules that include fats, waxes, sterols, fat-soluble vitamins such as vitamins A, D, E, and K, monoglycerides, diglycerides, triglycerides, phospholipids, and others. In this experiment lipid was extracted from the egg yolk of a chicken egg. Three forms of lipid were extracted namely total lipid, neutral lipid and polar lipid. Then using a square chromatoplate the lipid was subjected to a two-dimensional thin layer chromatography to separate the lipid into different components. Theoretically the egg yolk should contain TAGs, cholesterol ester, cholesterol, free fatty acids and polar lipids. In the end of the experiment the chromatoplate should show the polarity of the spots based on the adsorption of the spots in the mobile phase of the two-dimensional thin layer chromatography. Proper handling of the chromatoplate and developing chamber is recommended to avoid errors in the result.
INTRODUCTION
Lipids are substances found in living organisms that are insoluble in water but soluble in non polar solvents and solvents of low polarity. This lack of solubility in water is an important property because our body chemistry is so firmly based on water. Most body constituents including carbohydrates which are soluble in water. But the body also needs insoluble compounds for many purposes, including the separation of compartments containing aqueous solutions from each other, that’s where lipids come in.
The water-insolubility of lipids is due to the fact that the polar groups they contain are much smaller than their alkane-like (nonpolar) portions. These nonpolar portions provide the water-repellent, or hydrophobic, property (1).
An important use for lipids, especially in animals, is
Myca Pua, Ramon Ramos, Amanda Redilas, Kimleigh Reyes,
Nathaniel Sim and Clara Tamondong
Group 9 2F Medical Technology Biochemistry Laboratory
ABSTRACT
Lipids constitute a group of naturally occurring molecules that include fats, waxes, sterols, fat-soluble vitamins such as vitamins A, D, E, and K, monoglycerides, diglycerides, triglycerides, phospholipids, and others. In this experiment lipid was extracted from the egg yolk of a chicken egg. Three forms of lipid were extracted namely total lipid, neutral lipid and polar lipid. Then using a square chromatoplate the lipid was subjected to a two-dimensional thin layer chromatography to separate the lipid into different components. Theoretically the egg yolk should contain TAGs, cholesterol ester, cholesterol, free fatty acids and polar lipids. In the end of the experiment the chromatoplate should show the polarity of the spots based on the adsorption of the spots in the mobile phase of the two-dimensional thin layer chromatography. Proper handling of the chromatoplate and developing chamber is recommended to avoid errors in the result.
INTRODUCTION
Lipids are substances found in living organisms that are insoluble in water but soluble in non polar solvents and solvents of low polarity. This lack of solubility in water is an important property because our body chemistry is so firmly based on water. Most body constituents including carbohydrates which are soluble in water. But the body also needs insoluble compounds for many purposes, including the separation of compartments containing aqueous solutions from each other, that’s where lipids come in.
The water-insolubility of lipids is due to the fact that the polar groups they contain are much smaller than their alkane-like (nonpolar) portions. These nonpolar portions provide the water-repellent, or hydrophobic, property (1).
An important use for lipids, especially in animals, is
References: [1] Bettelheim,F.A., March,J. (1990). Introduction to organic and biochemistry. Philadelphia: Saunders College. [2] McKee. (2003). Biochemistry: The Molecular Basis of Life. Boston: McGraw-Hill. [3] Lehninger, A.L. (2008). Legninger Principles of Biochemistry. New York: W.H. Freeman. [4] Heftman, E. (1967). Chromatography. New York: Reinhold Publishing Corporation Polar lipids Figure 4 Theoretical Two-dimensional TLC Chromatoplate