Rapid liquid chromatographic-mass spectrometric assay for oxymetazoline in whole rat blood.

A rapid HPLC-electrospray mass spectrometric assay for the quantitation of oxymetazoline in whole rat blood has been developed. Sample preparation was a single liquid-liquid extraction after addition of a deuterated internal standard (IS) and pH adjustment. An aliquot of reconstituted extract was injected onto a narrow-bore octadecyl reversed-phase column at a flow-rate of 400 microliters/min. Using a 20:1 post-column split, 5% of the eluent was introduced into the mass spectrometer interface. Elution of the analyte and IS occurred in less than 2 min. This rapid separation was made possible because of the sample cleanup and the selectivity of the mass spectrometric detection. The [M+H]+ ions for oxymetazoline (m/z 261) and [2H9]oxymetazoline (m/z 270) were detected using selected ion monitoring. The linear range of the assay was 0.67-167 ng/g of blood and the limit of quantitation with a 0.30-g sample was 1.0 ng/g. The assay permitted the analysis of nine samples per hour with the requisite sensitivity and selectivity and was used to determine the blood pharmacokinetics of oxymetazoline in rats dosed via intravenous and intranasal routes.