Extraction of Spinach Lab Report OChem
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Extraction of Spinach Lab Report OChem

Course Number: OCHEM 241A/B, Fall 2011

College/University: Arizona

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6/14/11 Abstract The objective of this experiment was to isolate different fractions of spinach components using column chromatography and analyze the different fractions using Thin Layer Chromatography and obtain Rf values. The different pigments present in the spinach sample were Chlorophyll b, Xanthophyll, and Carotenes. The chlorophyll b had an Rf value of .44 and .06, xanthophyll had an Rf value of .38, and...

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objective 6/14/11 Abstract The of this experiment was to isolate different fractions of spinach components using column chromatography and analyze the different fractions using Thin Layer Chromatography and obtain Rf values. The different pigments present in the spinach sample were Chlorophyll b, Xanthophyll, and Carotenes. The chlorophyll b had an Rf value of .44 and .06, xanthophyll had an Rf value of .38, and the carotenes had an Rf value of .71. Introduction The objective of this experiment was to isolate different fractions of spinach components using column chromatography and analyze the different fractions using Thin Layer Chromatography and obtain Rf values. Column chromatography is an efficient way to separate the pigments because by sending solvents with different polarities through the column, the pigment attracted to the solvents polarity will be extracted and taken out of the column. The hexanes will be used to extract the least polar pigment because it is very non polar. The mix of 75/25 hexane/acetone will extract the slightly polar because it is slightly polar. Methanol will be used to extract the most polar out of the column because it is very polar. Thin Layer Chromatography in this experiment is a method that essentially compares the relative polarities of all the different pigments by obtaining Rf values. The Rf value is the distance the pigment travels divided by the distance the solvent travels. The higher the Rf value the less polar the pigment. The most polar pigments will not travel up the plate very far because they will stick to the silica coating of the TLC plate. The least polar molecules will travel farther up the TLC plate because it will be carried up the plate by the solvent. Discussion Through column chromatography Thin and Layer Chromatography, the pigments in the spinach were easily extracted and the Rf values were calculated. The pigments present in the spinach sample were Chlorophyll b, Xanthophyll, and Carotenes. The Rf value chlorophyll b was .44 and .06, the Rf value of Xanthophyll was .38, and the Rf value of Carotene was .71. When comparing the structures of the different pigments, the results that were obtained seem accurate. In the structure of chlorophyll, there are several sites for Hbonding, which will make the molecule polar. Also, the central bonds are four nitrogen atoms bonded to a much less electronegative magnesium atom. The dipoles of the nitrogen atoms will make the center more electron deficient, making it more prone to make new bonds. Because of this, it is safe to conclude that chlorophyll is the most polar pigment in the spinach sample. The second most polar pigment in the spinach sample was xanthophyll. When analyzing its molecular structure, this makes sense because there are two alcohol groups (O-H) on the molecule, which makes the molecule prone to H-bonding with other molecules. Xanthophyll is more polar than carotene because carotene is made of nonpolar alkane groups (C-H & -CH3). Because of this, carotene is the least polar pigment extracted from the spinach sample. The first sample taken had both chlorophyll b and carotene in in because when the hexane was added, the carotene was very hard to see. Because it was not detected, there was a large amount of excess hexane. This affected the results on the TLC plate in such a way that there were no spots visible on the plate. This is the part of the experiment could have been repeated in order to obtain an accurate reading of the pigments that were mistakenly mixed.

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