Plant Pigment Separation Lab

Plant Pigment Separation Lab - Conclusion: Isolation and...

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Conclusion : Isolation and Purification of Plant Pigments Drew Rasmussen 9/08/2011 This experiment was used to show students how to properly set up and “pack” a microscale chromotography column for the separation of low volatility compounds (such as the pigments extracted from the spinach). In addition to learning microscale chromatography techniques, students were introduced to Thin Layer Chromotography where a TLC plate could be read and measured to determine if two compounds had properly separated by analysis of their R f values. As part of the interpretation of the R f values, the different marks on the TLC plate was compared to determine whether full separation of the different compounds had occurred. First, a hot plate was obtained to begin boiling water for the spinach leaf pigment extraction process. During this time, a “Williamson microscale column” was set up for chromatography. The glass column with a micro Buchner funnel attached was “packed” with a slurry of anhydrous alumina and hexane as a nonpolar solvent. The column was filled about 2/3 of the way full with a protective barrier of washed sand on top of the alumina layer. Pigment was extracted from the boiled spinach leaves using by pulverizing the leaves and then rinsing with acetone. A dark green liquid came off of the leaves, and this was used as the “original sample” to separate the pigments out of. This was done by introducing the sample to the column and adding a solvent. The solvent started off as a 90:10 hexane acetone solution for the yellow
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This note was uploaded on 01/18/2012 for the course CHEMISTRY 308 taught by Professor Jonscaggs during the Fall '11 term at Boise State.

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Plant Pigment Separation Lab - Conclusion: Isolation and...

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