The application of high pressure liquid chromatography to the analysis of clinically important porphyrins

The porphyrias are a group of acquired or inherited metabolic diseases characterised by the abnormal production and excretion of porphyrins and porphyrin precursors. The clinical evaluation and subsequent treatment of porphyria are dependent on the analysis of these compounds in urine, feces and blood because a wide array of overlapping physiological manifestations make direct clinical diagnosis difficult. The analytical techniques currently available are inefficient and do not have the capacity to separate or easily quantify complex mixtures of porphyrins. High pressure liquid chromatography (HPLC), on the other hand, can readily achieve complex separations and since the sample being analyzed remains in solution at all times its detection and quantitation are considerably simplified. Bearing these considerations in mind, we have developed new porphyrin isolation techniques and evaluated a variety of HPLC packing/ solvent systems for the analysis of the uro to proto porphyrins from urine and feces. The HPLC procedures are rapid and efficient and should be easily extended to other porphyric samples. The versatility of the HPLC system was demonstrated by the characterization of copper coproporphyrin from a porphyric fecal sample. We have also developed procedures for the qualitative analysis of the fecal sub-uro porphyrins and made prelminary progress toward the separation and characterization of this heterogeneous group of compounds.Science, Faculty ofChemistry, Department ofGraduat