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Clinical Chemistry 34: 2481-2485, 1988;
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Clinical Chemistry, Vol 34, 2481-2485, Copyright © 1988 by American Association for Clinical Chemistry

Ferrochelatase activity in human lymphocytes, as quantified by a new high-performance liquid-chromatographic method

E Rossi, KA Costin and P Garcia-Webb
Clinical Biochemistry Department, Queen Elizabeth II Medical Centre, Nedlands, Western Australia.

We describe a new rapid, sensitive high-performance liquid- chromatographic (HPLC) method for determining ferrochelatase (EC 4.99.1.1) activity in lymphocytes. Zinc and mesoporphyrin are incubated aerobically with sonicated lymphocytes and the zinc mesoporphyrin formed is extracted with dimethyl sulfoxide-methanol-EDTA for quantification by HPLC. Incubation conditions, including the concentration of the palmitic acid activator, were optimized. The Michaelis constant (Km) was 2.1 mumol/L for mesoporphyrin, 22.2 mumol/L for zinc. The mean ferrochelatase activity (expressed as zinc mesoporphyrin formed per hour per milligram of lymphocyte protein) for a reference population was 3.25 (SD 0.43) nmol.h-1.mg-1. For three patients with erythrohepatic protoporphyria (EHP), activities were 1.11, 1.30, and 1.35. Neutrophils contain negligible ferrochelatase activity.


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