In Figure 4.

1, paper chromatography was used to separate the different

photosynthetic pigments from San Francisco leaves. In this part of the experiment,
9:! Petroleum:ether was used as the mobile phase where the mobility due to
solubility of the extracted photosynthetic pigments were based. Results showed 4
photosynthetic pigments namely: yellow green, green, yellow and orange. From
these results, we can say that the pgments yellow green and green were the least
soluble and the pigments yellow and orange were the more soluble ones. Also, we
can now say the identity of the pigments. Yellow green and green pigments are
most likely chlorophyll pigments while the other two are carotenoids or
xanthophylls.
Based from the data gathered in the spectroscopy of the photosynthetic
pigments, the absorption spectrum was made. The relationship betwee the
absorbance of the different pigments and wavelength was determined to be able to
visualize their absorbance peaks. Results showed that the yellow green pigment
peaked at 400 nm with absorbance of 0.135. for the green pigment, it peaked at
425 nm at 0.047. for the orange pigment, it peaked at 650 nm with absorbance of
0.19. lastly, the yellow pigment peaked at 475 nm with absorbance of 0.107. with
these given absorbance values, we can now determine the identities of the said
pigments. In order to identify the pigments, the wavelength for the thighest
absorbance of each pigment. The peak of the spectrum would indicate the
maximum absorbance of the pigment in that certain wavelength. According to
published literature, most carotenoids has a maximum absorbance value of 450
500 nm. In this case, we conclude that the yellow pigment was a carotenoid since it
peaked at 475 nm. The yellow and the orange pigment along with the yellow green
and green pigments have small difference in the absorbance values and we ca say
that the identity of these pigments are the same. In this experiment, the values
suggest that the yellow green and green pigments are most likely chlophylls.
Specifically, the yellow green pigment would be chl b and the green pigment id chl
a.
Figure 4.2 shows the standard curve made from the data gathered in the
absorption of different concentrations of Methylene Blue. The calculation of the
concentration of the unknown substance was done using the Lambert beer law. With
given concentrations and absorbance values, we were able to compute for the
concentration of the unknown using the formula for c. the calculation shown in the
worksheet is near to the interpolated value, this supports that the computed
concentration from the Lambert Beer law is correct. Based from these results we
can now say that the absorption and the concentrations of given samples have a
direct relationship. As the concentration of the sample increases, the value of
absorbance also increases. At higher concentration, there is more electrons,
therefore the higher the amount of light the pigments are able to absorb. Hence,
higher concentrations of the sample result to higher absorbance value.
In order to compute for the concentration of a certain substance given that it
initially registers at a high value and have a reading of 100%, one should dilute the
sample first. With this, the number of particles detected by the spectrophotometer
can be reduced and the initial reading would decrease. Also, the dilution should be

considered and should have an absorbance value that falls under the Lambert-Beer
plot that has been constructed for comparison. After getting the absorbance of the
diluted sample, we can now multiply the dilution factor from one of the points in the
plot to be able to get the exact concentration of the unknown.