The determination of histamine in challenged human leukocyte preparations by high-performance liquid chromatography

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L LeinoL Lauren


A highly sensitive and rapid method was developed for the determination of histamine in challenged human leukocyte preparations by high-performance liquid chromatography. The assay is based on the Shore's OPT-reaction of the unpurified sample and on a specific separation of the derivate with analytical reversed phase phenyl column combined with spectrofluorometric detection. The detection limit of histamine by this method was 0.07 pmol (signal to noise ratio 2:1) and the within-day variation for peak height was 3.6% and for retention time 0.8%. A good linear standard curve ranging from 12.5 pg to 500 pg (0.07 pmol-2.7 pmol) was obtained with correlation coefficient of 0.998. The histamine release from human basophils in mixed leukocyte preparation was induced by the calcium ionophore A 23187. A concentration of 0.4 micrograms/ml ionophore was required for 50% histamine release with a Ca2(+)-concentration of 1.8 mmol/l. The measured total histamine content was 1.5 pg/basophil.


Nov 1, 1986·Allergy·F EbbesenI Søndergaard
Jan 1, 1985·International Archives of Allergy and Applied Immunology·G Marone
Jan 1, 1987·International Archives of Allergy and Applied Immunology·T IshizakaH Saito
Jul 1, 1987·Analytical Biochemistry·T YoshimuraM Kimura
Apr 8, 1973·Life Sciences. Pt. 2: Biochemistry, General and Molecular Biology·D P EvansD S Thomson


Dec 2, 1994·Journal of Chromatography. B, Biomedical Applications·D EggerL Hültner
Aug 26, 2006·Immunology and Allergy Clinics of North America·Mariana Castells

Related Concepts

High Pressure Liquid Chromatography Procedure
Histamine Release

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