Studies on the isolation of plant pigments from Spinach leaves

ravikumar velpula


The leaves of plants contain a number of colored pigments generally falling into two categories, chlorophylls and carotenoids. The green chlorophylls a and b, which are highly conjugated compounds capture the light energy used in photosynthesis. Carotenoids are part of a larger collection of plant-derived compounds called terpenes. These naturally occurring compounds contain 10, 15, 20, 25, 30 and 40 carbon atoms, which suggest that there is a compound with five carbon atoms that serves as their building block. Their structures are consistent with the assumption that they were made by joining together isoprene units, usually in a "head to tail" fashion. Isoprene is the common name for 2-methyl-1,3-butadiene. The branched end is the "head" and the unbranched is the "tail". That isoprene units are linked in a head to tail fashion to form terpenes is known as the isoprene rule. Carotenoids are tetraterpenes. Lycopene, the compound responsible for the red coloring of tomatoes and watermelon, and β-carotene, the compound that causes carrots and apricots to be orange, are examples of carotenoids. β-Carotene which cleaves to form two molecules of vitamin A when it is ingested, is the major dietary source of this vitamin.

Spinach leaves, which you will use in this assignment, contain chlorophyll a and b and β- carotene as major pigments as well as smaller amounts of other pigments such as xanthophylls. The xanthophylls, which are oxidized versions of carotenes and pheophytins, look like chlorophyll except that the, Mg+2 is replaced by two hydrogen ions. In this assignment you will isolate and separate the plant pigments using differences in polarity to effect the separation. Since the different components are colored differently, the separation is easily followed visually.


Isolation, Extraction, Column Chromatography, Thin-layer chromatography

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