Normal-phase HPLC Separation of Possible Biosynthetic Intermediates of Pheophytin a and Chlorophyll a

• Published in: Analytical Sciences 2001, Vol.17(4), pp.509-513
• Main Authors: NAKAMURA, Akimasa, TANAKA, Shuhei, WATANABE, Tadashi
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Carbohydrate Sequence; Chlorophyll; Chlorophyll - analogs & derivatives; Chlorophyll - biosynthesis; Chlorophyll - isolation & purification; Chlorophyll - metabolism; Chlorophyll A; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; Heterocyclic compounds, pigments; Hordeum - chemistry; Isomerism; Liquid chromatography; Other biological molecules; Pheophytins - biosynthesis; Pheophytins - isolation & purification; Pheophytins - metabolism; Plant Leaves - chemistry; Plant Proteins - isolation & purification; Tetrapyrrolic pigments
• ISSN: 0910-6340; 1348-2246
• DOI: 10.2116/analsci.17.509

Normal-phase HPLC conditions have been developed for separating the C173 isoprenoid isomers, which are expected to be formed as biosynthetic intermediates of chlorophyll (Chl) a, Chl a′ (C132-epimer of Chl a), pheophytin (Pheo) a and protochlorophyll (PChl). The application of these conditions to pigment composition analysis of greening etiolated barley leaves allowed us to detect, for the first time, the C173 isomers of Chl a′, a possible constituent of the primary electron donor of photosystem (PS) I, P700, and those of Pheo a, the primary electron acceptor of PS II, in the very early stage of greening. The C173 isomer distribution patterns were approximately the same between Chl a and Chl a′, but significantly different between Pheo a and Chl a′, probably reflecting the similarity and difference, respectively, in the biosynthetic pathways of these pigment pairs.