New ultra-micro high-performance liquid chromatographic method for determining the γ chain composition of hemoglobin F in normal adults

F. Kutlar, T. H.J. Huisman

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

The difficulty in isolating the minute quantity of Hb F (<1%) present in the red blood cells of normal adults greatly complicates the determination of its γ chain composition. We have developed a rapid ultra-micro high-performance liquid chromatographic (HPLC) method and used it to analyze the γ chain composition of Hb F in 47 adults with Hb F levels between 0.1-3.4%. The method involves the isolation of Hb F from as little as 50 μl of whole blood on an analytical size cation-exchange HPLC column, followed by concentration in a Centricon micro concentrator unit and by reversed-phase HPLC analysis. The entire procedure can be completed in one day and 3-4 analyses can be made simultaneously. We reanalyzed the blood samples from 22 subjects with known β-globin gene cluster haplotypes, and confirmed the association of high, low, and very low Gγ levels with haplotypes A, B, and C, respectively. Also included are the results of DNA sequence analyses of the Gγ and β promoters, and of the locus-control-region hypersensitive site-2 (LCR-HS-2) of the β-globin gene cluster in five subjects homozygous for haplotypes A, B or C; the data obtained failed to provide a satisfactory explanation for all the variations in the Gγ levels that have been observed.

Original languageEnglish (US)
Pages (from-to)183-189
Number of pages7
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume620
Issue number2
DOIs
StatePublished - Oct 29 1993

ASJC Scopus subject areas

  • Chemistry(all)

Fingerprint Dive into the research topics of 'New ultra-micro high-performance liquid chromatographic method for determining the γ chain composition of hemoglobin F in normal adults'. Together they form a unique fingerprint.

  • Cite this