A rapid and fully-automated method for the quantitation of tricyclic antidepressants in serum using turbulent-flow liquid chromatography-tandem mass spectrometry.
BACKGROUND
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.


METHODS
Human serum and an internal standard 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 analytes were 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 ionized and detected over a 3.5 min 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.


RESULTS
The simple and complex precision CVs for all compounds were < or = 16%. The limits of detection and quantitation for all drugs were < or = 3 ng/ml and <20 ng/ml, respectively. Recoveries were between 97 and 114%. Slopes for method comparison plots were all >0.96. Proficiency testing materials had values within 2 SDI of peer group means for all drugs.


CONCLUSIONS
Based on validation data, this is a specific, sensitive fully-automated method for rapid quantitation of tricyclic antidepressants in serum.
Breaud
Autumn R
AR
The Johns Hopkins School of Medicine, Department of Pathology, Clinical Chemistry, Baltimore, MD, USA.
Harlan
Robert
R
Di Bussolo
Joseph M
JM
McMillin
Gwendolyn A
GA
Clarke
William
W
D000929:Antidepressive Agents, Tricyclic / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods / Q000592:standards; D006801:Humans; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards*; D013997:Time Factors
D000929:Antidepressive Agents, Tricyclic