Add time:07/25/2019         Source:sciencedirect.com

A sensitive method for the simultaneous determination of doxepin and its active metabolite DESMETHYLDOXEPIN (cas 1225-56-5) in plasma was established, using high-performance liquid chromatographic separation with tandem mass spectrometric detection. The samples were extracted with hexane–isoamyl alcohol, separated on a Phenomenex Luna C18 5 μm, 150×2.1 mm column with a mobile phase consisting of methanol–water–formic acid (600:400:0.5, v/v) at a flow-rate of 0.25 ml/min. Detection was achieved by a Perkin-Elmer API 2000 mass spectrometer at unit resolution in multiple reaction monitoring mode monitoring the transition of the protonated molecular ions m/z 280.2, 266.2 and 250.1 to the product ions m/z 107.1, 107.1 and 191.0 for analyte, metabolite and internal standard (benzoctamine-HCl), respectively. TurboIonSpray ionisation was used for ion production. The mean recovery for doxepin and desmethyldoxepin was 90% and 75%, respectively, with a lower limit of quantification at 0.320 ng/ml and 0.178 ng/ml for the analyte and its metabolite, respectively, using 0.5 ml plasma for extraction. This is the first assay method described for the simultaneous determination of doxepin and desmethyldoxepin in plasma using LC–MS–MS. The method is sensitive enough to be used in drug bioavailablity studies with doxepin.

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