CHEMICAL CHARACTERIZATION LIPID

AndradeS,Granados L.X
Corporacin tecnolgica de Bogot y Tecnologa en Regencia de farmacia, Bioqumica
Bogot D.C. Colombia
26 de Mayo de 2016

ABSTRACT
The egg contains lecithin, a capable
substance to connect to the cholesterol
to impede that this is totally absorbed
by our organism. The lecithin is in
natural way in our body, in the bone
marrow, in the brain, in the liver and in
the heart and is essential for the good
functioning of the nervous system.
Besides this substance helps to hold
concentration, an agent is, being
therefore known as,, a corazn.La's
important
protectress
thin-layer
chromatography is principally used for
separation, this technique is based
principally in the displacement for
capillarity in the plate and drag along
the components through this, the
separation takes place for the
differential migration, since the movable
phase drags the apolar substances and
that one that they are polar they are
retained. For the separation of the
present lipids in the huevo.Las try of
lipids they consist in drawing a
comparison every one of the solutions
prepared from the egg with the
corresponding lipids to identify if the
egg has lipids like lecithin and or
cholesterol.

INTRODUCTION
Cholesterol is a sterol (lipid) that is found
in body tissues and blood plasma of
vertebrates. Occurs
in
high

concentrations
in
the liver, spinal marrow , pancreas and b
rain. They are abundant in the fats of
animal origin. The chemical formula of
cholesterol is represented in two ways:
C27H/c
or4627H45OH. It
is
a lipid steroid, molecule of pentane
perhydro-phenanthrene
cycle (or esterano), consisting of four
carbocycles condensed or cast, called
A, B, C and D, presenting several
substitutions:
1. Two methyl radicals in positions C-10
and C-13.
2. A chain aliphatic branched
carbons in position C-17.

of

3. A hydroxyl group at the c-3 position.

Introduction between
4.unaC-5 and C-6.

the

carbon

On the cholesterol molecule you can

see
a polar
head made
up
of
the hydroxyl group and a tail or non
polar formed by the condensation nuclei
carbocyclic
portion
and
the
substituents aliphatic. Thus, cholesterol
is
a
molecule
so
hydrophobic
cholesterol free water solubility is 10 8
M and, like the other lipids, it is quite
soluble in polar solvents such
as chloroform (CHCl3).

MATERIALS AND METHODS

In a camera chromatography positioned
themselves approximately 5mL of a
solution of n hexane: Ethyl ether:
Glacial acetic acid (70: 25: 1) and let
himself be saturated the camera during
approximately 20 minutes.
2g of yolk of a soft boiled egg prepared
a sample placing in a mortar
approximately itself, they added 10mL
of a solution of ethanol: Ether:
Chloroform (2:1:2), macerated smoothly
and it filtered through filter paper,
collecting the masking in a glass of
precipitated.
The solvent on electric iron of heating
evaporated, stirring the glass constantly,
to have the viscous or dry abstract
without quitting coming to boiling of the
solvent
since
ether
is
highly
inflammable. Once I mark effectively
when defending in sports, himself
redisuelvi LM of ether evidences it in
10, and 30 added LM of acetone
themselves. It took shape one
precipitated, which let itself be decanted
placing the glass of precipitated in
diagonal way. He took his liquid phase
and he placed himself in a glass of
precipitated of 50 LM, got warm in
electric iron of heating to reduce the
volume to 10 LM (Solution 1).
Himself I let the solid phase in the
electric iron of heating mark effectively
when defending in sports, and himself
redisolvi the solid in 10 LM of ethyl
ether (Solution 2).
1 separated 1 LM of solution itself and
of solution 2 for the chromatography.
For the qualitative tests of composition
of lipids; They took in three different test

tubes, 1 LM of solution 1, of solution 2

and of solution of lecithin (correct
check). 1 LM of ammonium molybdate
and 1 LM of sulfuric acid concentrated
slowly by the walls of the tube were
added to each tube; It shook and he let
himself be rested for 3 minutes next 1
LM of ascorbic acid was added. In a
fourth test tube molybdate, sulfuric acid
and ascorbic acid were added
(continuity failure). 1, of solution took 1
LM of solution 2 and of glycerine
(correct check). A small quantity from
KHSO4 was added to the three
previous pipes and to a fourth empty
tube (continuity failure). Each tube to
the flame got warm lightly and the smell
unfastened in each tube was taken. 1,
of solution took 1 LM of solution 2 and
of solution of cholesterol (correct
check). They added 3 FeCl3's LM, and
1 LM of sulfuric acid once the three
previous pipes was concentrated and to
a fourth empty tube (continuity failure).
In the separation of lipids for thin-layer
chromatography;
On
the
chromatographic plate they measured
themselves with a pencil smoothly
leaving a 1-CM distance from the base
of the plate. With a capillary positioned
himself on the plate you show them
according to the following information:
Plate 1: Line 1: Solution 1. Line 2: Boss
of Cholesterol. Line 3: Boss of Lecithin.
Plate 2: Line 1: Solution 2. Line 2: Boss
of Cholesterol. Line 3: Boss of Lecithin.

The plate placed itself in the camera, it

got plugged and the passage of time
kept passing grade the plate until the
movable phase came to 1 CM of the top
of the plate. The plate was withdrawn

and the zone got marked with pencil as

far as the movable phase came. He let
himself be dried the plate, and himself I
apply
the
disclosing
solution
(Ammonium Sulphate 10 %). He let
himself be marked effectively when
defending in sports and positioned
himself the plate on an electric iron of
heating to 70 C approximately and he
left once they started to reveal colors on
this to him.

6
0,9
9
4
Board 1. Displacements in TLC, (sln1
solution 1, sln2 solution 2, BC boss
cholesterol, BL boss lecithin).
1

Image 2. Result of the plates

chromatography (PlacaIzquiera Sln1,
PlacaDerecha Sln2).

RESULTS
Thin-layer chromatography

Total
solve
nt
distan
ce
Dista
nce
sampl
e
(cm)
RF

Sln1

BC

BL

Sln
2

BC

BL

3,3

3,3

3,3

3,2

3,2

3,2 DISCUSSION OF RESULTS

1,3
3,0
0,39
0,91

1,6
1,9
3,0

0,4
1,5
2,2
3,1

0,4
8
0,5
8
0,9

0,1
2
0,4
5
0,6

1,5
2,9
0,4
7
0,9
1

0,2
1,
5
3,
0
0,0
6
0,
47
0,

The traveled distance for the solvent

1, came from 3,3cm for the plate of the
solution 1. The solutions 1 and 2
2,0 contain the sample, that he presents
3, separations, distinguished for the color
and traveled distances.
0,
The solution 1 and 2 originating of the
0,6 egg yolk has a refractive index in some
0,
similar points to the one of the boss of

cholesterol shown in bold in the tabla1

(1.5 and 3.0) of the as it can be said
that solutions have tall content of
cholesterol in their composition.
From the lecithin in the solution 2
refractive indexes look closer although
for very little as compared with the
solution 1, which indicates that the
solution 2 can have a bigger degree of
quantity of lecithin in their composition
without than the solution 1 accent of
having her because values as we said
previously are very close.
From the obtained data we can
corroborate the presence of cholesterol
and lecithin in the egg yolk.
The lipid fraction of the yolk through
maceration was carried out in a medium
solvent (ethanol: ether: chloroform) in
proportions 4:2:4 being these of
different polarities since ethanol is a
polar solvent, petroleum ether is nonpolar and chloroform is fairly polar in
order to separate lipids present
breaking the covalent bonds. To filter
this solution evaporates it to remove the
solvent present in solution taking
appropriate care since it is volatile. To
get extract redissolved in ether and
acetone, ether is a solvent of low
polarity and under 35 C. [1] Boiling
point what makes it special for lipids,
resins and mineral fats therefore will
contain fat and cholesterol while
acetone as a solvent intermediate
allows miscibility two solvents that are
not
and
the
extraction
of
substances This will contain these
phospholipids. A precipitate after the
rest was formed at the edge of the
glass, to the separate phases are
prepared solution (reducing to 10 mL) 1
and solution 2 (by adding 10 mL of ethyl

ether to the solid) that is used for thinlayer chromatography.

Figure # 1 Solutions 1 and 2

Qualitative identification tests of the

composition of lipid in egg yolk by
coloration, precipitation and aromaticity
tests were made.
The first test was performed by me
method Lieberman- Burchard with
solution 1, 2, lecithin, ascorbic acid,
ammonium molybdate and [H2SO4] for
the positive control which winch to a
blue coloration and evidenced a
precipitation in the lower part of the tube
being similar to solution 1 while in the
negative control solution present a
green coloration. Which tells us that the
egg yolk contains lecithin Phospholipid
Figure # 2 controls lecithin

Was the test of aromaticity with KHSO4 ,

solution 1, solution 2 and Glycerin was
used for the positive test to pass tubes
through the flame they presented a
smell similar to the smell of the punch

with the exception of the second tube

does not present this scent

which indicates to us that the egg yolk

contains cholesterol.

Figure # 3 tests KHSO4

Figure # 4 tests for cholesterol

BlBLIOGRAPHY

Test for cholesterol with solution 1,

solution 2, FeCl3 and sulphuric acid in
which positive test was visibly very
similar to solution 1 a bit more light

LEHNINGER.
PRINCIPIOS
DE
BIOQUMICA /Michael M. Cox, David
L. Nelson/ EdicionesOmega/ 1264