Determination of hydroxy and peroxy acid derivatives of uroporphyrin in the plasma of patients with congenital erythropoietic porphyria by high-performance liquid chromatography.

A reversed-phase high-performance liquid chromatographic method is described for the determination of hydroxy and peroxy acid derivatives of uroporphyrin in the plasma of patients with congenital erythropoietic porphyria. The porphyrins were extracted from the plasma with 20% trichloroacetic acid-dimethyl sulphoxide (1:1, v/v). The supernatant after centrifugation was chromatographed on a Hypersil-ODS column by gradient elution with 9% (v/v) acetonitrile in 1 M ammonium acetate buffer (pH 5.16) (solvent A) and 10% (v/v) acetonitrile in methanol (solvent B) as the gradient mixture. The method was also suitable for the preparative isolation of the porphyrins.