Ion-pair reversed-phase high-performance liquid chromatography of algal chlorophylls

The chromatographic behaviour of algal chlorophylls under ion-pair reversed-phase liquid chromatography conditions has been tested either on monomeric and polymeric octadecylsilica columns. Three five-carbon amino compounds (pentylamine, piperidine and pyridine), differing in their molecular shapes and chemical properties, have been used as counterions. The monomeric octadecylsilica column did not allow the separation of chlorophylls c1 and c2 with any amine, but separated chlorophyllide a, chlorophyll c3 and Mg-divinylpheoporphyrin a5 when pyridinium acetate was used as ion pair agent. The polymeric octadecylsilica column separated chlorophylls c1 and c2 with the three counterions, achieving the highest retention and overall separation of acidic chlorophylls when pyridinium was employed as counterion.