Simultaneous separation and quantification of free and metalchelated protoporphyrins in blood by three-dimensional HPLC. Clin Chem

In this new method for simultaneous separation and quan-tification of free and metal-chelated porphyrins in blood, the porphyrins are extracted from blood samples with a mixture of diisopropylamine:water:methanol (25:100:900, by vol) and separated by HPLC elution. The data are collected in three-dimensional form with a microcomputer. This method permits a high recovery of protoporphyrin (PP) and Zn-chelated PP as well as heme from blood. By this method, the amounts of these analytes in erythro-cytes from normal or abnormal subjects can be deter-mined more accurately than by conventional methods. IndexIng Terms: erythrocytes/heme/zinc protoporphyrin/three-di-mensional spectra: fluorescence (or absorbance) vs wavelength time Porphyrins exist in human blood and tissues in bot