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  Home > JCE Print > Journal of Chemical Education > Issues > 2004  > March  >
In the Laboratory
An Improved Method for the Extraction and Thin-Layer Chromatography of Chlorophyll a and b from Spinach
Hao T. Quach, Robert L. Steeper, and G. William Griffin
Science Department, Bunker Hill Community College, Boston, MA 02129

Cover
March 2004
Vol. 81 No. 3
p. 385

Abstract
A direct method of extracting plant pigments from spinach leaves into a dry organic solvent is presented. A powder is obtained by grinding a mixture of raw spinach, drying agent, and sand (1:1:2) that can be extracted directly into acetone in 10 minutes to provide a sample suitable for chromatographic analysis. This method avoids liquid–liquid extractions and subsequent drying as is found in previously reported methods. Removal of the magnesium ion from chlorophyll pigments was observed by treating the acetone extract with a strong ion-exchange resin for three minutes. The loss of magnesium ion is often observed as a degradation product of chlorophyll samples. An improved solvent system consisting of petroleum ether, cyclohexane, ethyl acetate, acetone, and methanol in 60%, 16%, 10%, 10%, and 4% quantities, respectively, is also reported. TLC separation using this extraction method and solvent system allows for the clear separation of chlorophyll a and b from all other extracted pigments.

Featured on the Cover

Supplement
A copy of our laboratory handout and a list of chemicals and materials used in this experiment are available. We have also attached copies of figures that show how this method works for a number of green vegetables and extracts from dried and fresh spinach stored for extended periods of time to show the pigment degradation products.
*  Contents
*  Download
JCE2004p0385W.pdf

More Information
*  Citation
Quach, Hao T.; Steeper, Robert L.; Griffin, G. William. J. Chem. Educ. 2004 81 385.
*  Keywords
Chromatography; General Chemistry; Introductory / High School Chemistry; Laboratory Instruction; Natural Products; Plant Chemistry; Separation Science
*  History
Created:
Last Updated:
February 3, 2004
February 18, 2005
Link to Cover added (April 2004).
  Home > JCE Print > Journal of Chemical Education > Issues > 2004  > March  > Page 385


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