The group's objectives are to separate

colored components of siling labuyo using

column chromatography and to determine the
purity of the components using thin layer
chromatography. The group also aim to
measure the retention factor values of each
colored components of siling labuyo obtained
from thin layer chromatography.

MATERALS AND METHODOLOGY
n performing column and thin layer
chromatography, the group members used siling
labuyo as their sample. The experiment could be
performed with the use of capillary
tubes,Pasteur pipette, mortar and pestle,
beaker, test tubes, iron stand, iron clamp, filter
paper, and precoated TLC plates. Silica gel was
used as an adsorbent in performing the column
chromatography. DCM Hexane was used as the
solvent for the system. DCM, DCM-hexane
(1:1), and DCM: Methanol were used as the
eluents in the separation of the components of
siling labuyo.
or the experiment proper, first the
group triturated five pieces of siling labuyo then
they extracted its pigments using DCM-hexane
(1:1). They had set aside a portion of the
extracts, to be used for thin layer
chromatography. They uniformly packed the
column with silica gel up to the indented part of
the Pasteur pipette. They placed 0.5 mL of the
extract on top of the column. The group then
eluted the pigment mixture using 10 mL of DCM-
hexane (1:1). They introduced the solvent
system, or the eluent, in portions. The group
used DCM-hexane (1:1), DCM, and DCM:
methanol as their eluents. The column should
not run dry. They discarded the colorless eluate
and collected the colored eluates in separate
test tubes.

igure 1: The process of eluting the pigment
mixture using DCM.
The group then performed the Thin Layer
Chromatography by applying the eluates on a
precoated TLC plate by spotting each eluate 10
times at a marked origin an inch from the bottom
of the TLC plate. Each spot was allowed to dry
before applying the next.

igure 2: Spotted TLC plate.
The developing chamber was then prepared
by placing approximate amount of the solvent
system, DCM-hexane (1:1), on a beaker. A filter
paper was lined on the inner wall of the chamber
then it was covered with watch glass and was
allowed to equilibrate. The plate was carefully
placed in the developing chamber. The solvent
was allowed to rise up to 1 cm from the upper
end. The plate was then removed from the
chamber and the solvent front was then
immediately marked and air dried. To ensure an
approximate marking on the solvent front, the
group visualized the components using a UV
lamp. The retention factor values was then
computed followed by the documentation of the
chromatoplates.


igure 3: Thin layer chromatographer set up;
TLC plate spotted with an eluate is placed inside
a developing chamber and was covered.
RESULTS AND DSCUSSON
Upon performing the column
chromatography, the group was able separate
the components of siling labuyo. They were able
to gather four different colored components of
siling labuyo and these are yellow, orange, red
orange, and red with their respective volume in
drops.
Table 1: CoIumn Chromatography ResuIts
Color of Component Volume of eluate
(drops)
1 Yellow 2 drops
2 Orange 1.30 mL
3 Red Orange 1.20 mL
4 Red 3 drops


igure 4: The different eluates collected from
siling labuyo through the use of column
chromatography.
The purity of the components were
determined using the thin layer chromatography.
The group was able to obtain the retention factor
value of each component using the distance of
the component from the origin marked on the
precoated TLC plate. The retention factor value
was computed by dividing the distance travelled
by the compound over the distance travelled by
the solvent The distance of the solvent is 6.3
cm from the point of origin.
Table 2: %hin Layer Chromatography ResuIts
Color of
component
Distance of
component
from origin
(X) in cm
Rf Value
1 Yellow 4.1 cm 0.65
2 orange 4.4 cm 0.70
3 Red
orange
5.1 cm 0.81
4 red 3.2 cm 0.51


REERENCES:
O www.chemguide.co.uk/analysis/chromat
ography/thinlayer.html
O Raymond P. W. Scott. Chrom-Ed book
series, Thin layer chromatography.
(2007)
O Norton, Peter B., Esposito, Joseph J.
(1994) Chromatography, The New
Encyclopedia Britannica. Chicago. P.
286