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Ther Drug Monit. 2002 Aug;24(4):537-44.

Simultaneous determination of seven tricyclic antidepressant drugs in human plasma by direct-injection HPLC-APCI-MS-MS with an ion trap detector.

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  • 1Institute of Forensic Medicine, Karl-Franzens University Graz, Graz, Austria.


A sensitive and specific direct-injection high-performance liquid chromatography-atmospheric pressure chemical ionization tandem mass-spectrometry (HPLC-APCI-MS-MS) method has been developed for the rapid identification and quantitation of seven tricyclic antidepressants-amitriptyline, nortriptyline, doxepin, dosulepin, dibenzepin, opipramol, and melitracen-in human plasma. After the addition of the internal standard lofepramine and dilution with 0.1% formic acid, plasma samples were injected into the LC-MS-MS system. Proteins and other large biomolecules were removed during an on-line sample cleanup using an Oasis extraction column (1 x 50 mm, ID, 30 microm) with a 100% aqueous mobile phase at a flow rate of 4 mL/min. The extraction column was subsequently brought in-line with the analytical column by automatic valve switching. Analytes were separated on a 5-microm Symmetry C18 (Waters) analytical column (3.0 x 150 mm, ID) using a step gradient of acetonitrile-0.1% formic acid at a flow rate of 0.6 mL/min. The total analysis time was only 12 minutes per sample. The interday and intraday coefficients of variation for all compounds were </=11%. The assay was sufficiently sensitive to monitor all seven compounds within their therapeutic ranges. The proposed method has been in routine use for more than 1 year and permits direct analysis of plasma samples without time-consuming sample preparation, which may be especially advantageous for clinical laboratories.

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