SEPARATION OF CHLOROPHYLL-A AND METALLOPORPHYRINS BY HPLC IN NORMAL-PHASE MODE USING A DIOL-PHASE

The isolation of high-purity chlorophyll a was achieved by HPLC using a micro-preparative column with a diol-phase (Polyol, Si 100, 5 mum) i n normal-phase mode. The procedure included the extraction of photosyn thetic pigments from the leaves of stinging nettles and their partial purification as dioxane-water-chlorophyll-aggregates prior to the isol ation by micro-preparative HPLC. Freeze-drying resulted in a flaky chl orophyll a, which was free of water. Each step of isolation was contro lled by using an analytical HPLC-column with a diol-phase and a photod iode array detector. Synthesized metalloporphyrins (metal(II)-ion = Mg 2+, Zn2+, Cu2+, Ni2+) with structures analogous to chlorophyll were al so isolated by means of the micro-preparative HPLC-column containing a diol-phase. The purity of the obtained porphyrins was monitored with an analytical HPLC-column.