Chlorophyll Analysis by New High Performance Liquid Chromatography Methods

Abstract

The improvements in high performance liquid chromatography (HPLC) analysis of chlorophylls (Chls) and bacteriochlorophylls (BChls) during the last decade rely mainly on the application of newly developed stationary phases combined with new mobile phases developed with special regard to the nature of the ionpairing agents employed for achieving the retention of free acid forms, which is especially important for the Chls c group. The application of mass spectrometry (MS) as a detection technique coupled on-line with liquid chromatography (LC) has provided important structural information. All these tools have contributed to the discovery of the biosynthetic pathways of higher plant Chls, to the study of the composition and distribution of new Chls in algae, and to the characterization of complex pools of BChls. Faster, more sensitive and more specific HPLC methods are expected in the very near future, with the development of new columns and detection techniques, e.g. monolithic stationary phases and coupled liquid chromatography-nuclear magnetic resonance (LC-NMR) systems.