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Separation of mono- and divinyl chlorophyll species by high-performance liquid chromatography using an octadecyl
Y Shioi1, K Watanabe, K Takamiya
1Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Abstract:
Most of the chlorophylls and their related compounds from nonesterified chlorophyllide to esterified pheophytin were separated by high- performance liquid chromatography using a wide pore, C(18) polyvinyl alcohol polymer column with an elution using a binary gradient starting from a buffered mobile phase. The high selectivity of this system enabled not only the separation of common chlorophyll species but also the resolution of structurally similar compounds such as mono- and divinyl chlorophyllide and pheophorbide species which usually coelute on monomeric bonded phases. The method is successfully applied to analyses of the pigments extracted from green and etiolated tissues of higher plants and photosynthetic bacterial cells.