Observations on the use of o-phthalaldehyde condensation for the measurement of histamine.
Siegel-PD; Lewis-DM; Petersen-M; Olenchock-SA
Analyst 1990 Aug; 115(8):1029-1032
The measurement of histamine was used as a quantitative marker in an in-vitro investigation of mast cell degranulation. This investigation addressed the reaction conditions and the stability of the o-phthalaldehyde (OPT)/histamine adduct and further evaluated the purity of leukocyte histamine extracts. The formation and degradation of the adduct was measured at both 0 and 25 degrees-C under alkaline conditions. The OPT/histamine adduct was observed to be very stable at 0 and 25 degrees-C, following acidification. Derivatization conditions and the purity of the leukocyte histamine extract were assessed and indicated that derivatization at low temperatures slowed down decay, and provided a greater overall fluorescence intensity. The results demonstrated the necessity of an extraction procedure for the isolation of histamine from leukocyte supernatant fluids. The authors state that the use of butanol in the extraction procedure can cause some quenching of the fluorescent product. However, it has been demonstrated to be a reliable solvent for most leukocyte histamine extraction purposes.
NIOSH-Author; Analytical-methods; Chemical-analysis; Analytical-chemistry; Cell-function; Cell-biology; Temperature-effects; Chromatographic-analysis