RESULTS AND DISCUSSION

Color of Component

In this experiment, two types of chromatography

Volume of Eluate
(Drops)

were used for the purpose of separation and

1
2
3

purity determination. Column chromatography,

These eluates were then used as individual

which runs a solvent that moves down slowly

samples for spotting the TLC plates which were

through a glass column with packed adsorbent,

then put inside the developing chamber for

was used to separate the colored components of

completing the thin layer chromatography. The

malunggay leaves. On the other hand, thin layer

movement of these eluates along the TLC plate

chromatography (TLC) which utilizes a stationary

coated with silica-gel were possible through the

phase composed of a thin layer of adsorbent

principle of capillary action.[1] Retention factors

material on a glass plate or plastic sheet, was

were then determined by measuring the distance

used

each

of each component from the origin and by

collected eluate to overall determine their purity

dividing this to the solvent front, represented by

based

the formula:

based on solid-liquid chromatographic methods

to

separate

on

the

the

components

retention

values

of

(Rf

values)

obtained.
Starting

with

separating
malunggay

the

column

chromatography

for

colored

components

the

leaves,

hexane-acetone

of

was

determined to be the suitable solvent system to

be used as it run through the glass column with
packed adsorbent without dissolving the solid.

Distance of sample
Distance of solvent
Rf = origin origin

Table 2. Characteristic Color and Distance of

Component from Origin (X) with Determined
Rf Value
Distance of

This reagent was continuously added up on the

column to prevent it from running dry, as the
pure extract of malunggay leaves was prepared
with the same reagent, to be added next to
perform the chromatography process.
As the pure extract slowly moved down the
column, 3.0 mL each of the solvents, specifically
hexane-acetone, acetone, and acetone-methanol
respectively, were added continously at the same
time while collecting different colors of eluates in
separate test tubes. For this, four differentcolored eluates were collected, of which two were
mixed to provide three distinct-colored eluates,
particularly, pale yellow, green, and yellow green.
The following table presents the data collected
under column chromatography.
Table 1. Characteristic Color and Volume of
Collected Eluate (Column Chromatography)

Pale yellow
Green
Yellow Green

1
2
3

Color of

component

Component

from origin (X)

Pale Yellow
Green
Yellow Green

in cm
4.89 cm
4.79 cm
0.42 cm

Rf Value

0.75
0.74
0.42

After the Rf values were calculated, several

deducations were made. The difference in the Rf
values of the components are based from their
solubility and the strength of their adsorption to
the adsorbent, in the case the TLC plate coated
with silica gel. The lower the Rf value of the
component, the more soluble it is to the solvent
system used, which was hexane-acetone. This is
based on the solubility principle like dissolves
like. The more soluble the component in the
solvent system is adsorbed more firmly to the
TLC plate.[1] With this principle, it was deduced

that the third component is non-polar, like the

polarity of the solvent sytem hexane-acetone.
From the Rf values calculated, it is also possible
to determine the identity the component by
comparing the experimental Rf values with the Rf
values of known components. The spots on the
TLC plate is also an indicator of purity although it
is not an absolute indicator

of it. A pure

compound would appear as a single spot on the

plate

and

impurities.

several

[2]

spots

are

indications

of

It is also deduced that how close the

Rf values of a component to a known compound

is an indication of the purity of the component.
These

explanations

correlated

with

obtained

the

is

Figure 1. Chromatogram under UV light

now

results

from

the

chromatogram shown below.

The third component has a
polarity alike with the solvent
system, which was non-polar,
since it had the lowest Rf
value

out

of

components.

the
The

three
several

spots on the Chromatogram,

specifically

in

the

second

component

is

also

an

Figure 2. Dried Chromatogram with

Markings

indication

that

the

components we have separated still contain some

impurites. These impurites can be from the
different apparatus used and even from the
different

solvents

Chromatography.

used

in

Column

REFERENCE
From the Internet
[1] University of Massachusetts. (n.d.). Thin
layer Chromatography [PDF]. Retrieved from
http://www.chem.umass.edu/~samal/269/tlc.pdf
[2] Interpretation of experimental data [PDF].
(2011, August). Retrieved from
http://chemistry.syr.edu/totah/che276/support/6
a1.handouts/4.DataInterp.pdf