RESOLUTION OF LUTEIN AND ZEAXANTHIN USING A NON-ENDCAPPED, LIGHTLY CARBON-LOADED C-18 HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC COLUMN

A new rapid and reproducible high-performance liquid chromatographic method using Spherisorb ODS-1, a non-endcapped, lightly carbon-loaded column material, for the separation of higher-plant chloroplast pigments is described. The method resolves lutein and zeaxanthin, as well as all other major and most minor pigments at or near baseline by either of two solvent programs. Program I is faster and more sensitive than program II while the latter resolves pheophytin a and beta-epsilon-carotene slightly better than program I. Both programs use an initial buffered aqueous mixture that appears critical for this application of ODS-1. The method is well suited for analysis of xanthophyll-cycle pigment changes.