Quantification of Tricyclic Antidepressants in Serum Using Liquid Chromatography Electrospray Tandem Mass Spectrometry (HPLC-ESI-MS/MS)

  • Christopher A. Crutchfield
  • Autumn R. Breaud
  • William A. ClarkeEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1383)


Tricyclic antidepressants (TCA) are used to treat major depressive disorder and other psychological conditions. The efficacy of these drugs is tied to a narrow therapeutic window. Inappropriately high drug concentrations can result in serious side effects such as hypotension, tachycardia, or coma. As a result, concentrations of tricyclic antidepressants are routinely monitored to ensure compliance and to prevent adverse side effects by dose adjustments. We describe a method for the determination of concentrations of amitriptyline, desipramine, imipramine, and nortriptyline in human serum using high-performance liquid chromatography coupled to a tandem mass spectrometer with electrospray ionization (HPLC-ESI-MS/MS). The method is rapid, requiring only 3.5 min per analysis. The method requires 100 μL of serum. Concentrations of each TCA were quantified by a calibration curve relating the peak area ratio of each TCA analyte to a deuterated internal standard (amitriptyline-D3, desipramine-D3, imipramine-D3, and nortriptyline-D3). The method was linear from ~70 ng/mL to ~1000 ng/mL for all TCAs, with imprecision ≤ 12 %.

Key words

Tricyclic antidepressants Depression Tandem mass spectrometer Amitriptyline Desipramine Imipramine Nortriptyline 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Christopher A. Crutchfield
    • 1
  • Autumn R. Breaud
    • 1
  • William A. Clarke
    • 2
    Email author
  1. 1.Department of PathologyThe Johns Hopkins University School of MedicineBaltimoreUSA
  2. 2.Department of Pathology-Clinical ChemistryThe Johns Hopkins University School of MedicineBaltimoreUSA

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