Extraction, purification, and characterization of chlorophylls from spinach leaves
Khalyfa, A. | Kermasha, S. | Alli, I.
Crude chlorophyll was extracted from fresh spinach leaves by use of methanol and acetone as two separate solvents. The crude chlorophyll was partially purified by successive precipitation using a mixture of dioxane and water. The partially purified chlorophyll was subjected to column chromatography with DEAE-Sepharose CL-6B. Chlorophylls a and b were obtained by further purification of a Sepharose CL-6B column chromatography. The recovery of chlorophyll fractions was determined, and the degree of their purity was demonstrated by spectrophotometric scanning and by HPLC analysis. The extraction of 100 g of fresh spinach leaves with methanol and subsequent purification procedures resulted in 38 mg of chlorophyll a (30.1% purity) and 13 mg of chlorophyll b (27.5% purity). Extraction with acetone and subsequent purification procedures resulted in 61 mg of chlorophyll a (44.5 % purity) nd 29 mg of chlorophyll b (32.4% purity), respectively. The purities of commercial samples of chlorophylls and b were 36.0% and 32.4%, respectively.
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