High-performance liquid chromatographic determination of zinc protoporphyrin and porphyrin carboxylic acids in urine.
A method for the reversed-phase high-performance liquid chromatographic separation of zinc protoporphyrin and porphyrin carboxylic acids with fluorescence detection and its application are described. A mu Bondapak C18 column was employed for all the experiments in this study. The method required a pretreatment of the column with a two-component mobile phase containing 0.1 M NaH2PO4 in acetonitrile (28:15, v/v, pH 5.3) for 10 min prior to sample injection. Separation was achieved isocratically by increasing the concentration of acetonitrile in the mobile phase (0.1 M NaH2PO4:acetonitrile, 18:130, v/v, pH 5.3) 4 min after injection to complete the elution. The flow rates and the period of pretreatment of the column were studied to optimize the separation. The method was applied to determining zinc protoporphyrin and porphyrin carboxylic acids of heme biosynthesis in urine.
Ho
J W
JW
Department of Pediatrics, State University of New York, Buffalo 14214.
D002264:Carboxylic Acids / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005470:Fluorometry / Q000379:methods; D007855:Lead Poisoning / Q000652:urine; D011166:Porphyrins / Q000652:urine*; D011524:Protoporphyrins / Q000652:urine*
D002264:Carboxylic Acids; D011166:Porphyrins; D011524:Protoporphyrins; C017803:zinc protoporphyrin