Isolation of Chlorophyll and Carotenoid Pigments from Spinach

Isolation of Chlorophyll and Carotenoid Pigments from Spinach

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Isolation of Chlorophyll and Carotenoid Pigments from Spinach Observations: I enjoyed this experiment. Although, I thought we could had run more efficiently, so that it ran a lot faster and smoother. Results: There was a great deal of spotting with the TLC plate. We were able to determine Carotenes (1 spot yellow-orange), Pheophytin a (gray), Pheophytin b, Chlorophyll a (blue-green, more intense than chlorophyll b), Chlorophyll b (green), and Xanthophylls (yellow, there spots). Conclusion: I think we did very well considering the time it took to get our results. We had a hard time heating the substance so that we can evaporate the solvent in the test tube, which took about 30 minutes. We also used the wrong absorbent in part 1 of this experiment, we used alumina instead of anhydrous sodium sulfate, and someone decided to keep the sulfate with them at their lab bench. Questions:
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Unformatted text preview: 1.The chlorophylls are more polar than the carotenes so they adhere more strongly to the alumina in the column, thus their slower mobility. The same principle applies to their movement up the TLC plate. Their slower mobility up the plate causes them to not travel as far as the carotenes when development is stopped. Since the further the substance travels the higher the Rf value, their Rf value is lower than that of the carotenes. 3. Rf values would be higher because Acetone is more polar than hexane, the components would move faster across the TLC plate. We used 70%hexane-30% acetone for preparing our development chamber 4. There were yellow and green bands in the column but other colors showed up during TLC development. Some additional components aside from the yellow and green ones can result from air oxidation, hydrolysis, and other chemical reactions....
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