Abstract
A nano-liquid chromatographic /tandem mass spectrometric (nanoHPLC/MS/MS) method has been developed for the sensitive determination of histamine, a vital neurotransmitter. The method involved pre-column derivatization of histamine with 7-fluoro-4-nitrobenzoxadiazole (NBD-F), thus increasing the hydrophobicity of the analyte and allowing it to be retained and stacked on a nanoelectrospay emitter packed with C18 reversed-phase particles. Sodium 1-heptanesulfonate, added as an ion-pairing reagent in the mobile phase for sample loading, was found to facilitate greatly the analyte retention. The packed nanoelectrospray emitter was easily prepared using fused-silica capillary of 75 microm i.d., and included creating an electrospray emitter tip and then slurry-packing a 5 cm long column with 5 microm silica-based particles. After sample loading, the mobile phase was changed for separation and a characteristic precursor-to-product ion transition, m/z 275 --> 258, was used for the quantification. A linear calibration curve in the range from 1.0-100 ng/mL histamine with an r2 value of 0.9995 was obtained. The detection limit was estimated to be 0.1 ng/mL histamine in water. In a preliminary validation, the method was applied to determine histamine in alcoholic beverages and rat brain tissues.
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