Column Chromatography of Lipids

Chromatography of lipids using a glass column filled with a suitable material is a common
and useful method for fractionation of lipid classes either on an analytical or a semipreparative scale. The retention results in a variety of mechanisms including hydrogen
bonding, Van der Waals' forces and also ionic bonding. The solid phase is relatively polar
(normal chromatography) and the more polar the lipid, the more strongly is it adsorbed.
Thus, the lipids are eluted by increasingly polar solvents. This technique has a low resolution
when used at low pressure (Solid Phase Extraction or SPE) but has a high resolution (high
performance) when run at high pressure using a stationary phase made of fine particles
(HPLC). The former is restricted to the fractionation of complex mixtures into two or three
less complex ones, the later being adopted to analyze and quantify purified fractions.
Extraction from the chicken egg yolk are separated based on its differences in solubility.
There are factors affecting lipid solubility like chemical nature of the molecule, atomic or
molecular formula weight, valence or charge or sphere of hydration, charge density and
sphere of hydration. Also, in analyzing the lipids present in the crude extract using column
chromatography, it is necessary to first isolate them quantitatively from nonlipid
components. Extraction of lipids from source materials, such as food, animal and plant
tissues,
or microorganism, should be carried out in a manner that avoids changes in the lipids or
leads to the formation of artifacts it has eluates and each was differentiated by its
components.
The first eluate must be a triacylglycerol or triglyceride for (9:1) mixture of petroleum
ether:ethyl ether. Triacylglycerols are the main components of animal and plant lipids. They
are an ester of three fatty acids and glycerol. They are the most concentrated source of
energy in the human body and are stored in subcutaneous fat deposits where they
contribute to insulation. Fat deposits contain over 70 000 kcal of stored energy, but the
triacylglycerol is not immediately accessible for muscle respiration because it must be
broken down into its basic components for transport in the blood and then oxidized before
entry into the krebs cycle. Triglycerides have lower densities than water (they float on
water), and at normal room temperatures may be solid or liquid. When solid, they are called
"fats" or "butters" and when liquid they are called "oils".
The second eluate was cholesterol for 5% methanol in dichloromethane. Cholesterol is a
chemical compound that is naturally produced by the body and is a combination of lipid (fat)
and steroid. Cholesterol is a building block for cell membranes and for hormones like
estrogen and testosterone. About 80% of the body's cholesterol is produced by the liver,
while the rest comes from our diet. Cholesterol is only one of several lipids (fats) circulating
in our blood stream. Its components, Triglycerides are an additional form of fat (3 fatty
acids plus glycerol) circulating in the blood. Cholesterol and Triglycerides cannot dissolve in
water due to being lipids, or fats. Because our blood is comprised primarily of water, for
Cholesterol and Triglycerides to circulate through your blood, the Cholesterol and
Triglycerides must be carried by protein packages called Apoproteins.
The third eluate must be lecithin for dichloromethane:methanol:water (1:3:1). Lecithin is
a lipid that consists mostly of choline, but also includes inositol, phosphorus, and linoleic
acid. Lecithin helps to prevent arteriosclerosis, protects against cardiovascular disease,
improves brain function, helps keep the liver and kidneys healthy, aids in thiamin and

vitamin A absorption, and can even help to repair liver damage caused by alcoholism, this
nutrient is essential to every living cell in the human body.