3

Figure 1. Paper Chromatography of the Pigments in Iresine herbstii

As can be observed in the figure presented above, three different pigments were separated
during the process of paper chromatography. The pigment which travelled the farthest is green and the
one lagging behind is red-purple and the yellow-green pigment comes in the middle. This indicates that
the red-purple pigment is not very soluble in the hydrophobic solvent; it possesses a greater affinity for
the hydrophilic paper. While the green pigment is the most soluble to the non-polar solvent followed by
the yellow-green pigment since they travelled the farthest. These observations are consistent with the
data presented in the table below.

Table 1. Distance Travelled by Pigments

Band Number Distance (mm) Solvent Front (mm) Band Color Rf Value
1 21 70 Red-Violet 0.3
2 30 Yellow-Green 0.43
3 45 Green 0.64

Results showed that bands one, two, and three, travelled 21 mm, 30 mm, and 45 mm across the
chromatogram paper, respectively, while the solvent travelled 70 mm. The Rf value for band one, which
is 0.3, is close to the standard Rf value range, which is 0.32-0.62, for monoglycosylated anthocyanin.
Therefore, it can be concluded that band one is monoglycosylated anthocyanin, also, these pigments
migrate slowly in non-polar solvent, which was why it lagged behind the other two pigments.

The Rf value computed for band number two is 0.43, it is near the standard Rf value for
chlorophyll b, which is 0.42. This indicates that band two is chlorophyll b. For band number three, the
computed Rf value is 0.64. It is close to the standard Rf value of chlorophyll a, which is 0.59. This implies
that the band which travelled the farthest is chlorophyll a. As stated in the introduction, chlorophyll a is
soluble to non-polar solvents, which was why it moved fast across the chromatogram paper.

Several errors may have occurred during the conduction of the experiment which led to the
discrepancies between the obtained Rf values in the experiment with that of the standard Rf values for
chlorophyll a, chlorophyll b, and anthocyanin. One source of error was that the paper used in the
experiment was an unsterile filter paper which may have been exposed to contaminants which can
affect the rate of movement of the pigments across it.