Chlorophyllides and pheophorbides were separated by ion-suppression high-performance liquid chromatography using an octadecyl silica column eluted with 80-95% (v/v) methanol in water containing 13 mM acetic (final pH 4.2) as the suppessing ion. The separated pigments were detected fluorometrically with a sensitivity in the picomol range without artifact formation. This technique can be used not only for the determination of non-esterified chlorophylls, but also for esterified chlorophylls, thus enabling the simultaneous identification of chlorophylls and their derivatives.
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