Chromatography on Florisil in the quantitative estimation of urinary and other porphyrins

We describe the use of Florisil chromatography to purify and fractionate porphyrins into uro-, copro-, and protoporphyrins before their measurement by quantitative fluorometry. Porphyrinogens in fresh urine may be rapidly oxidized to porphyrins by chloranil, or slowly in weakly alkaline solution, before the sample is applied to the Florisil column. Aliquots of the separate ammoniacal eluates may be lyophilized for additional fractionation by direct thin-layer chromatographic analysis if desired. The behavior of delta-aminolevulinic acid, porphobilinogen, and other pyrrole compounds in this system is also described.