Abstract
A reversed-phase h.p.l.c. system is described for the separation of the four type-III heptacarboxylic porphyrinogen isomers. The effects of buffer concentration, pH and type and proportion of organic modifier in the mobile phase on retention and resolution of isomers were studied. Optimum separation on an ODS-Hypersil column was by elution with a ternary mobile phase of acetonitrile, methanol and 1 M-ammonium acetate, pH 5.16 (7:3:90, by vol.). Isomer identification was based on a comparison of their retention times with those of authentic standards, and was further confirmed by h.p.l.c. analysis of the characteristic mixture of three pentacarboxylic porphyrins formed after partial decarboxylation of individual isomers in 0.3 M-HCl at 160 degrees C.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Jackson A. H., Sancovich H. A., Ferramola A. M., Evans N., Games D. E., Matlin S. A., Elder G. H., Smith S. G. Macrocyclic intermediates in the biosynthesis of porphyrins. Philos Trans R Soc Lond B Biol Sci. 1976 Feb 5;273(924):191–206. doi: 10.1098/rstb.1976.0009. [DOI] [PubMed] [Google Scholar]
- Li F., Lim C. K., Peters T. J. Separation and characterization of pentacarboxylic porphyrinogen isomers by high-performance liquid chromatography with electrochemical detection. Biochem J. 1987 Apr 15;243(2):621–623. doi: 10.1042/bj2430621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lim C. K., Li F., Peters T. J. High-performance liquid chromatography of uroporphyrinogen and coproporphyrinogen isomers with amperometric detection. Biochem J. 1986 Mar 15;234(3):629–633. doi: 10.1042/bj2340629. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lim C. K., Rideout J. M., Wright D. J. High-performance liquid chromatography of naturally occurring 8-, 7-, 6-, 5- and 4-carboxylic porphyrin isomers. J Chromatogr. 1983 Dec 30;282:629–641. doi: 10.1016/s0021-9673(00)91640-6. [DOI] [PubMed] [Google Scholar]
- Smith S. G., Rao K. R., Jackson A. H. The prophyrins of normal human urine, with a comparison of the excretion pattern in porphyria cutanea tarda. Int J Biochem. 1980;12(5-6):1081–1084. doi: 10.1016/0020-711x(80)90216-5. [DOI] [PubMed] [Google Scholar]