Vivien Coombs

AP Biology Period 4

Schulte

Experiment #4

January 30-31, 2018

The Effect of Isopropyl on the Rate of Chromatography in Spinach to Determine the Use of

Plant Pigments in Photosynthesis

 Abstract

This experiment looks at the effects of isopropyl alcohol on spinach chromatography in

order to study the relation of plant pigments in photosynthesis. Pigment from the spinach leaves
was rolled onto coffee filter with a quarter and the paper was placed into green isopropyl alcohol,
oil, and distilled water. The coffee filter paper did not show any signs of results in both the water
and the oil; however, the isopropyl alcohol showed a small amount of growth of mostly blue
colored pigment up to and Rf value of .909. We then discarded the coffee filter paper with all
three of the solvents and proceeded to repeat the experiment using a thicker paper and only using
the isopropyl alcohol to see the chromatography. By doing so, a greater outcome was observed
and the various shades of blue and green could be seen, with some definite lines of a green and a
blue. The coffee filter paper would only show very faint lines as opposed to the definite lines on
the thicker paper.
The pigments of plants are determined by the colors they absorb; for example, most
plants are green or contain some amount of green because they mostly absorb colors such as red
and blue, but do not absorb any green making the plants reflect the green instead. Chlorophylls
are the most important pigments in photosynthesis and contain both chlorophyll ‘a’ and
chlorophyll ‘b’, colored blue-green and yellow-green respectively. The thicker paper we used
showed the movement of the solvent(isopropyl alcohol) up the paper, we then measured the
movement of the solvent and the pigment to determine the Rf value of the solvent (the distance
the pigment moved up divided by the distance the solvent moved up). The Rf value is the
Relative Mobility Factor, which measures the mobility of both the solvent and the pigment. The
hypothesis is that the thicker paper would produce a better range of color pigments than the
coffee filter do to it being more of a sponge-like base to better move the pigment line up the
paper.

Data

Solute Total Distance Total Distance Rf Value

Solvent Move Up Pigment Moved Up (D​pigment​/ D​solvent​)

Isopropyl Alcohol 11cm Chlorophyll ‘A’- 10cm Chlorophyll ‘A’-.909

(coffee filter) Chlorophyll ‘B’- 8.5cm Chlorophyll ‘B’-.773

Isopropyl Alcohol 15.5cm chlorophyll ‘A’- ​13.5cm Chlorophyll ‘A’-.871

(Thicker Paper) Chlorophyll ‘B’- 12cm Chlorophyll ‘B’-.774

This table does not include the oil and the distilled water since we have no data for those
solvents. The solvent distance for both chlorophyll ‘a’ and ‘b’ was the same in both of the
different solutes.
 Summary

We were able to get a much better result when we used the thicker paper than with the
coffee filter paper, which proved our initial hypothesis that we would see more pigments on thick
paper. While in the data table it shows that the Rf value of the chlorophyll ‘a’ was the highest at
.909, the pigment that was seen on the filter was not a strong pigment and could barely be seen
enough to measure. The isopropyl was also able to move up the thicker paper more than the filter
paper, proving its more sponge-like ability to hold up the solvent. From what colors were seen on
the thicker paper we are able to see that the spinach leaves we used, possessed mostly
chlorophyll ‘a’ and chlorophyll ‘b’. Both of these chlorophylls are the most important pigments
in photosynthesis; the chlorophylls transfer the lights they absorb and transfer it to the reaction
center of the cell to be used as energy for photosynthesis to occur. The experiment only really
worked with the thicker paper, but it was interesting to compare the two different papers and the
effect that the type of paper that was used had on the experiment; this experiment definitely
needed a thicker paper that could hold all of the solutions or else the results wouldn’t be
accurate.

Bibliography
What Are the Roles of Chlorophyll A & B? (n.d.). Retrieved February 07, 2018, from

https://sciencing.com/what-are-the-roles-of-chlorophyll-a-b-12526386.html