Simultaneous determination of histamine and prostaglandin D2 using an LC-ESI-MS/MS method with positive/negative ion-switching ionization modes: application to the study of anti-allergic flavonoids on the degranulation of KU812 cells

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Abstract
A new method for simultaneous determination of histamine and prostaglandin D2 (PGD2) by liquid chromatography–electrospray ionization tandem mass spectrometry operated in positive and negative ionization switching modes was developed and validated without a previous derivatization step. This method was used to measure histamine and PGD2 release following degranulation of KU812 human basophilic cells, using pyrazol and d4-PGD2 as internal standards. Analyses were performed on a liquid chromatography system employing a Cosmosil 5C18 PAQ column and an isocratic elution with mixed solution of methanol–water (7:3, v/v) with 0.0015% trifluoroacetic acid at a flow rate of 0.2 mL/min. A triple-quadrupole mass spectrometer operating in selected reaction monitoring mode simultaneously monitored using the following transitions: positive m/z 112/95 for histamine and negative m/z 351/271 for PGD2. The retention times of histamine and pyrazol were 4.2 and 5.0 min, respectively. PGD2 and d4-PGD2 had retention times of 8.5 min. The limits of detection were 0.3 and 0.5 ng/mL for histamine and PGD2, respectively. The relative standard deviations of the retention time and peak area for histamine were between 1.6% and 7.7%, and were 1.2% and 7.8% for PGD2. This method was used to evaluate the anti-allergic effects of 26 flavonoids and sodium cromoglicate which are first-line anti-allergic drugs. Of these compounds, baicalein and morin were the most potent inhibitors.

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