The Use of High Pressure Liquid Chromatography for the Identification and Preparation of Pigments Concerned in Photosynthesis

Abstract

The chlorophylls and carotenoids present in preparations from chloroplasts of marine algae can be extracted and separated by high pressure liquid chromatography. The reverse-phase columns that were used gave good separation of the different pigments. Addition of the ion-pairing agent tetrabutylammonium phosphate to the methanol/water solvents gave improved separations with complex mixtures, and identification was facilitated by examination of the column eluant at 440nm, where all the pigments absorbed, and at 650nm, where only chlorophylls absorbed. The method allowed the isolation of individual pigments for further study.