DETERMINATION OF PORPHYRINS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHYWITH CHEMILUMINESCENCE DETECTION

The peroxyoxalate chemiluminescence reaction. based on the oxidation o f bis (2,4 dinitrophenyl) oxalate with hydrogen peroxide, was develope d as a new detection method for the determination of porphyrins after separation by reverse phase high performance liquid chromatography. To optimize signal to noise ratio, experimental parameters such as desig n of the detection flow-cell and the effects of pH and organic solvent s were investigated. Significant enhancements in detectability, select ivity and linearity were demonstrated for the determination of various hydrophilic and hydrophobic porphyrins when compared to absorbance an d fluorescence methods (e.g., the detection limits (S/N = 3) for uropo rphyrin and coprophorphyrin were about 40 and 5 fmol, respectively). A pplication of this method for the sensitive and selective determinatio n of urinary porphyrins was also demonstrated.