Urine and faecal porphyrin profiles by reversed-phase high-performance liquid chromatography in the porphyrias

Abstract

A novel reversed-phase system is described for the simultaneous separation of type I and type III isomers of uro-, heptacarboxylic-, hexacarboxylic -, pentacarboxylic - and copro -porphyrins and the dicarboxylic meso- and proto-porphyrins. The porphyrins were resolved on a Hypersil -SAS column eluted with 10% (v/v) acetonitrile in 1 mol/l ammonium acetate, pH 5.16 (solvent A) and 10% (v/v) acetonitrile in methanol (solvent B) with a linear gradient from 100% A (0%B) to 35% A (65%B) in 30 min followed by isocratic elution at 65% B for a further 10 min. The method is simple and reproducible and has been applied to the analysis of porphyrins in urine and faeces from patients with acute intermittent, variegate, hereditary copro , congenital erythropoietic, erythrohepatic proto and symptomatic porphyrias.