The Isolation of Plant Pigments by Column and Paper Chromatography CHEM 2071 Sect-023 Felipe Shinsato Drawer: 107

In this lab, the paper chromatography of the solution containing pigments present in spinach leaves resulted in three distinct colors. The colors of the pigments present were yellow, a greenish-yellow, and a darker green. The first band yellow was most likely composed of betacarotene, and the second yellow band was composed of xanthophyll. The greenish-yellow pigment was most likely composed of chlorophyll a, and the darker green pigment was chlorophyll b. The pigments separated from one another due to differences in polarity. The filter paper was polar, so as the mobile phase traveled through the filter paper the more polar compounds began binding to the filter paper. Two distinct bands of yellow formed. However, nonpolar hexane easily dissolved the nonpolar carotene which allowed it to be easily separated from the polar molecules such as the chlorophylls and xanthophyll. Since, xanthophyll was next least polar molecule the second band of yellow formed near the green bands . Chlorophyll a being less polar than chlorophyll b traveled higher up the paper before binding to the water molecules on the filter paper. Chlorophyll b being the most polar travelled the least. The column chromatography yielded similar results. However, the colors of the fractions collected were yellow, green, and a pale greenish-yellow. The first fraction collected was yellow in color, probably a mixture of carotenes and xanthophyll. A green mixture followed which was most likely composed of the chlorophylls. The pale greenish-yellow fraction didnt have a direct relation with any pigment present in the spinach leaves . The mixture could be a product of chlorophyll b and some residual carotenes and xanthophyll.

Although the chromatography lab did not require complicated processes, it provided plenty of experimental data. For example, if only the colors of the pigments are known, the polarity of each pigment in relatively to one another could be determined . In the context of this lab, the pigments were identified since both the colors and relative polarities were known. This lab could have been improved through better extraction of the pigments present in the spinach. The spinach leaves were added too early to the water. To compensate for this error, the spinach leaves were boiled longer than the recommended time which could have resulted in pigments being lost to the water. A more concentrated solution of pigments would have resulted in more distinguished separations.

References: "Chromatography." Wikipedia. Wikimedia Foundation, 29 Sept. 2013. Web. 02 Oct. 2013.