Using a Microscale Approach To Rapidly Separate and Characterize Three Photosynthetic Pigment Species from Fern

A rapid separation of three photosynthetic pigments (chlorophyll a and b and xanthophyll) from fern (Polystichum acrostichoides) is described using microscale solvent extraction and traditional thin layer chromatography that minimizes use of harmful chemicals and lengthy procedures. The experiment introduces students to the concepts of extraction and separation of compounds. Combined with absorption and fluorescence spectroscopy using a plate reader, this laboratory experiment demonstrates the concept of light absorption by different pigments that are involved in photosynthesis as well as their photoluminescence properties. This new approach allows students to perform multilaboratory techniques within a 3 h laboratory session. The experiment has been used successfully in an introductory level course, but it may be easily adapted to second-year courses such as organic chemistry laboratory where these methods are widely used.