An Improved Method Fòr Extraction, Partial Purification, Separation and Isolation of Chlorophyll from Spinach Leaves

Powdered sugar column chromatographic separation and isolation of chlorophyll were studied. A wet column was employed using a Pharmacia R25 chromatographic tube connected, through a UV monitor, to a fraction collector. Chlorophyll extracted from spinach leaves with acetone was selectively precipitated with dioxane and water, and then was washed with 80% (v/v) aqueous methanol. Use of the further purified chlorophyll as a starting material for chromatographic separation and isolation of chlorophyll, made column chromatographic procedures become extremely effective and easier. Thin-layer chromatographic and high-performance liquid chromatographic tests revealed that chlorophyll preparations in this study did not contain any other photosynthetic pigments and their degradation products.