Analysis of Histamine by On-Column Derivatization

Histamine was analyzed by on-column derivatization using Asahipak ODP-50 4E. When a stimulation, enough to cause inflammation, is applied to mast cells, they release histamine and other physiologically active substances as a result of their cell degradation. The analysis of histamine is widely performed using mast cells by bio-assay to assess their inflammation and/or allergy.
This chromatograms show analysis of histamine released during the degradation induced by the compounds 48/80(A), mastoparan (B) and somatostatin (C). Histamine analysis is possible without the interference of coexisting substances.
Refer to Instruments for On-Column Derivetization

Sample : Histamine released from rat peritoneal mast cell inducted by (A),(B) and (C)

 Column : Shodex Asahipak ODP-50 4E (4.6mmID*250mm)
 Eluent : CH3CN - 50mM Sodium tetraborate(18:82,v/v) containing 1mM OPA(o-Phthalaldehyde) and 1mM
 NAC(N-Acetyl-L-cysteine)
 Flow rate : 0.5mL/min
 Detector : Fluorescence detector(Ex 340nm,Em 450nm)
 Column temp. : 40°C

K.Saito, M.Horie, N.Nose,K.Nakagomi and H.Nakazawa, J.Chromatogr., 595,163(1992)