22. A Rapid and Fully-Automated Method for the Quantitation of Tricyclic Antidepressants in Serum using Turbulent Flow Liquid Chromatography-Tandem Mass Spectrometry
Tue 4:30 PM - PosterSplash Track 3
Autumn Breaud
Johns Hopkins University
Autumn R. Breaud,The Johns Hopkins University
Robert Harlan, The Johns Hopkins University
Joe Dibussolo, ThermoFisher Scientific
Gwen McMillin, ARUP
William Clarke, The Johns Hopkins University
Here we describe a fully-automated turbulent-flow liquid chromatography-tandem mass spectrometry method for the detection of tricyclic antidepressant drugs (amitriptyline, desipramine, imipramine, and nortriptyline) in serum.

Human serum and an internal standard mixture were injected directly onto a Cyclone P online solid phase extraction (SPE) column (0.5 x 50 mm). Following removal of serum proteins and other components the sample was transferred to a Hypersil Gold C-18 (50 x 3 mm) analytical column. Elution occurred with a gradient of water and acetonitrile each with 0.1% formic acid. Analytes were then ionized and detected over a 3.5 minute analysis time by electrospray ionization mass spectrometry with selected reaction monitoring (SRM). Matrix effects were well-characterized and carryover, precision, linearity, recovery and limits of detection and quantitation were evaluated.

The inter-day and intra-day coefficients of variation for all compounds were < or = 12%. The limit of detection for all drugs was < or = 3 ng/mL and the limit quantitation for all drugs was <20 ng/mL. Recoveries were between 98 and 123% for all drugs. Slopes for method comparison were all >0.90 and proficiency testing materials had values within 1 SDI of peer group means for all drugs.

Based on the validation data, this is a specific and sensitive fully-automated method for rapid quantitation of tricyclic antidepressants in serum.