Kinetic determination of serum haptoglobin with a centrifugal analyzer

T. S. Kickler, P. F. Fong, G. F. Johnson, H. M. Solomon

Research output: Contribution to journalArticle

Abstract

We describe a method for determining haptoglobin with a centrifugal analyzer that is based on haptoglobin combining stoichiometrically with hemoglobin to form a complex that has peroxidase like activity proportional to the quantity of haptoglobin present. Under assay conditions, unbound hemoglobin exhibits only a small fraction of the total peroxidase activity. Activity is measured colorimetrically at 405 nm after reaction with o dianisidine and ethyl hydrogen peroxide. The procedure is standardized by saturating a known amount of hemoglobin with a serum whose hemoglobin binding capacity exceeds the amount of hemoglobin in the assay system. The mean and mean within run precision of our method, determined by performing 17 replicate assays of both a pooled normal serum and a 10 fold dilution of the serum, was 1.13 g/liter (CV, 2.9%), and 106 mg/liter (CV, 5.8%), respectively. The 95 percentile estimate of the normal range by our method is 0.45-1.85 g/liter hemoglobin binding capacity. When results by our automated method were compared to those by a manual method, the slope of the unweighted linear least squares regression line was .970, the y intercept 26 mg/liter, and the correlation coefficient .995.

Original languageEnglish (US)
Pages (from-to)1962-1967
Number of pages6
JournalClinical chemistry
Volume22
Issue number12
DOIs
StatePublished - 1976

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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