Unit

UNIT 8.3 Measurement of Heme Concentration

  1. Peter R. Sinclair1,2,
  2. Nadia Gorman2,
  3. Judith M. Jacobs2

Published Online: 1 MAY 2001

DOI: 10.1002/0471140856.tx0803s00

Current Protocols in Toxicology

Current Protocols in Toxicology

How to Cite

Sinclair, P. R., Gorman, N. and Jacobs, J. M. 2001. Measurement of Heme Concentration. Current Protocols in Toxicology. 00:8.3:8.3.1–8.3.7.

Author Information

  1. 1

    Veterans Administration Medical Center, White River Junction, Vermont

  2. 2

    Dartmouth Medical School, Hanover, New Hampshire

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: MAY 1999

Abstract

Heme (iron protoporphyrin IX) is a prosthetic group for a number of hemoproteins in different tissues (e.g., hemoglobin, myoglobin, cytochrome P-450s, mitochondrial cytochromes, catalases, and peroxidases). Mutations in the biosynthetic pathway can affect the synthesis and/or degradation of heme. Several assays are provided in this unit for quantifying heme: a spectophotometric assay based on the characteristic absorption spectrum of oxidized and reduced form of the hemochrome formed by replacing the nitrogen ligands with pyridine; a fluorescence assay based on removal of the iron by a heated, strong oxalic acid solution to produce fluorescent protoporphyrin; a reversed-phase HPLC assay to measure heme and intermediates in the synthetic pathway; and a radiometric assay to measure newly synthesized heme in tissue culture cells.