Detection of delta-aminolevulinic acid, porphobilinogen and porphyrins related to heme biosynthesis by high-performance liquid chromatography.

Two new high-performance liquid chromatographic methods are described for the quantitative determination of porphyrins and their precursors. In our method, sub-nanomole quantities of porphyrins, delta-aminolevulinic acid and porphobilinogen derivatized with ophthalaldehyde were injected onto a C18 reversed-phase column and eluted with 0.1 M monobasic sodium phosphate-methanol-tetrahydrofuran (4:6:3) and detected with a spectrofluorometer. A second reversed-phase system using methanol-tetrahydrofuran-22 mM acetate buffer (15:6:11) was also developed.