- Purification of Chlorophyll α
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Forms green crystals from Me2CO, Et2O + H2O, Et2O + hexane + H2O or Et2O +pentane + H2O. It is sparingly soluble in MeOH and insol in pet ether. In alkaline soln it gives a blue-green colour with deep red fluorescence. A very crude mixture has been purified by chromatography on low melting polyethylene (MI 0.044; 'Dow' melting index MI < 2) and developed with 70% aq Me2CO. The order of effluent from the bottom of the column is: xanthophylls, chlorophyll β, chlorophyll α, phaeophytins and carotenes. A mixture of Chlorophyll α (CAS NO. ) and β is best separated by chromatography on sugar and the order is chlorophyll β elutes first followed by chlorophyll α. To an Me2CO-H2O soln of chlorophylls 200mL of iso-octane is added and the mixt shaken in a separating funnel and the H2O is carefully removed. The iso-octane layer is dried (Na2SO4) and applied to a glass column (5cm diameter) dry packed with l000mL of powdered sucrose which has been washed with 250mL of iso-octane. Elution with 0.5% of isopropanol in iso-octane gives chlorophyll α. Keeping the eluate overnight at 0°C yields micro crystals which are collected by filtration or centrifugation (Yield 40mg).
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