Reversed phase high performance thin layer chromatography and column liquid chromatography of chlorophylls and their derivatives

Suzuki, N.; Saitoh, K.; Adachi, K.

Journal of Chromatography 408: 181-190


ISSN/ISBN: 0021-9673
DOI: 10.1016/s0021-9673(01)81800-8
Accession: 006337719

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The reversed-phase liquid chromatographic retention behaviour of chlorophylls-a and -b, pheophytins-a and -b and pheophorbides-a and -b has been studied on an octadecyl-bonded stationary phase with mobile phase solvents of different polarities, including alcohols, acetone, acetonitrile and mixtures of ethanol and water. The a type of each chlorophyll derivative shows greater retention than the b type in every solvent tested. In both a and b series, the retention increases in the following order: pheophorbide < chlorophyll < pheophytin. The retentions of pheophorbides-a and -b are enhanced specifically by addition of a salt, such as sodium chloride, to the mobile phase used. The successful HPLC separation of a mixture of chlorophylls, pheophytins and pheophorbides is demonstrated by using ethanol-water (95:5, v/v) containing 0.005 M sodium chloride as the mobile phase.