Development of an LC-ESI-MS/MS method for the determination of histamine: application to the quantitative measurement of histamine degranulation by KU812 cells.

A rapid, simple, and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed for the identification and quantification of histamine without a previous derivatization step or the addition of general ion-pairing reagents to the mobile phase. This method was used to measure histamine release following degranulation of KU812 human basophilic cells, using pyrazol as an internal standard. Analyses were performed on an LC system employing a Cosmosil 5C(18) PAQ column and an isocratic elution with methanol-0.005% trifluoroacetic acid (1:1) at a flow rate of 0.2 mL/min. A triple-quadrupole mass spectrometer, equipped with an electrospray ionization interface was employed, operating in the positive ion mode. The retention time of histamine and the internal standard were 4.0 and 5.0 min, respectively. The relative standard deviations (R.S.D.s) of the retention time and peak area were between 0.47% and 2.03%. Micropipette tip solid-phase extraction (SPE) using LooseTip C(18) allowed for not only rapid sample preparation, but also decreased suppression effects, improving peak shape. This method was used to evaluate the anti-allergic effects of compounds contained in Taxus yunnanensis extracts. Four constituents that were isolated from the wood extracts of T. yunnanensis and sodium cromoglicate, which is used as a first line anti-allergic drug, were tested in an in vitro histamine release inhibition assay. Of these compounds, taxiresinol and isotaxiresinol were more inhibitory than sodium cromoglicate.