Isolation of chlorophylls a and b from spinach by counter-current chromatography

A method for the isolation of chlorophylls from spinach by counter-current chromatography was developed. An initial extraction protocol was devised to avoid the notorious sensitivity of chlorophylls to degradation by light, heat, oxygen, acids and bases. Further purification and separation of chlorophylls a and b were achieved using counter-current chromatography. Chlorophyll structures and purities were established by HPLC, fast atom bombardment mass spectrometry and nuclear magnetic resonance. Purity was estimated to be >95% (100% by HPLC). Typical yields from 30 g of freeze-dried spinach were 300 mg of chlorophyll a and 100 mg of chlorophyll b.