Exercise 10

Thin layer chromatography is an essential technique in separating and

determining the purity or identity of lipids. TLC has a characteristic of having a high
sensitivity while separating the mixture firmly and in great speed. Non-polar
solvents ascends along the plate and is dependent on its affinity with the moving
phase and stationary phase. Thus, it can be said that the separation is dependent
on the difference in partition coefficient. (Crawford, 2013).

The retention factor (Rf) should be compared against the given set of
standards in order to determine the purity and identity of the lipid. Under the same
condition, a compound that has a larger R f value is said to be less polar because it
gets away from the stationary phase but if not, the it is more polar and most
probably have a lower Rf value (Clark, 2016).

distance travelled by the s a mple

distance travelled by the solvent
Rf = the origin theorigin

This exercise aims to understand the principles behind the thin layer
chromatography technique and to use it in identification of the major lipid fractions
from different sources such as egg yolk, margarine, and butter.

Clark, J. (2016). Thin Layer Chromatography. Retrieved on April 20, 2017 from
http://www.chemguide.co.uk/analysis/chromatography/thinlayer.html

Crawford, N ( 2013). Retention Facctor in Chromatography. Retrieved on April 20,

2017 from http://study.com/academy/lesson/retetion-factor-in-chromatography-
definition-formula-quiz.html
 Lipid, an organic molecule that has lower solubility/ insoluble in water. Lipids
can be separated from the tissue of the animals and plants with the use of hot
ethanol, ether, benzene, etc. Also, neutral fats is an example of a saponifiable lipid
and can be hydrolysed by heat and alkali. On the other hand, steroid, a non-
saponifiable lipid cannot be hydrolysed even by heating. Primary and secondary
alcohol groups, free and esterified carboxyl group, amine groups, etc. are just some
of the functional groups that can be found in lipids (Bioinfo. org , n.d.).

Lipids have extensive composition, thus, it makes the extraction difficult but
since some techniques are currently available, extraction is now possible to
perform. The two techniques that can be used in extracting lipids are the
chloroform-methanol (2:1) solvent system and the ethanol (3:1) solvent system. In
order to separate and purify lipids, column chromatography and special washers on
solvent must be used. Lastly, since polyunsaturated and monounsaturated fatty
acids are common in majority of lipids therefore to properly extract lipids, the
performer must be very cautious and must execute it an area with low temperature
(Bioinfo. org , n.d.).

This exercise aims to extract lipids and obtain its components through
different solvent systems.