Extraction, purification and characterization of chlorophyll a and chlorophyll b from some sources

In the first stage of this thesis, chlorophylls contained in powdered lyophilized spinach, spirulina and tomato stems were extracted using different solvents (acetone, methanol and ethanol). The results indicated that lyophilized spinach contained higher levels of chlorophyll than the other tested sources. Furthermore, considering the amounts of both chlorophyll a and chlorophyll b, acetone was determined to be the most suitable solvent for the extraction of chlorophylls. In the second phase of the thesis, advanced purification studies of chlorophylls extracted from lyophilized spinach powder using acetone were conducted. For this purpose, chlorophyll a, chlorophyll b, and other color pigments in spinach were separately obtained by a column chromatography technique using silica gel as a packing material. Chlorophyll a and b fractions obtained from column chromatography were applied to thin-layer chromatography using a mixture of petroleum ether:2-propanol:water (90.7:9.07:0.23, v/v/v) as the mobile phase. After separation of the pigments in the thin-layer chromatography, chlorophyll a and b were scraped from the plate and taken up in methanol before being filtered. High-pressure liquid chromatography was used to identify and determine the purity levels of chlorophyll a and b after purification. Our method yielded 0.075 mg of chlorophyll a with a purity of 91% and 0.0318 mg of chlorophyll b with a purity of 86% from 0.5 g of lyophilized spinach powder.