Systematic development of an UPLC–MS/MS method for the determination of tricyclic antidepressants in human urine

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 88; pp. 660 - 665
• Main Authors: Chambers, Erin E., Woodcock, Matthew J., Wheaton, Jessalynn P., Pekol, Teresa M., Diehl, Diane M.
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01-01-2014
• Subjects: Antidepressant; Antidepressive Agents, Tricyclic - analysis; Antidepressive Agents, Tricyclic - urine; Calibration; Chromatography, High Pressure Liquid; Depression - drug therapy; Depression - urine; Drug Monitoring - methods; Humans; Incurred Sample Reanalysis; Reference Standards; Reproducibility of Results; Solid Phase Extraction; Tandem Mass Spectrometry; Tricyclic; UPLC–MS/MS; Urine; Antidepressant; Urine; Incurred Sample Reanalysis; Tricyclic; UPLC–MS/MS
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2013.09.001

•The UPLC–MS/MS method measures five tricyclic antidepressants in human urine.•Simple screening protocols and subsequent optimization delivered the final method.•The method was evaluated against current FDA guidelines.•The work includes an assessment of incurred sample reanalysis (ISR).
Tricyclic antidepressants have been prescribed for the treatment of depression and other disorders since their discovery in the 1950s but have been replaced in recent decades by newer drugs with more favorable side effect profiles. However, for some patients and conditions, tricyclic antidepressants remain the drug of choice. A fast, sensitive, and robust UPLC–MS/MS method for the monitoring of amitriptyline, nortriptyline, imipramine, doxepin, and desipramine in human urine has been developed using a pre-defined and systematic method development approach. The method was developed using sub-2-μm particle technology, providing a state-of-the-art alternative to older methods. Total cycle time was 2.5min. Human urine samples (200μL) were prepared using an Oasis® WCX μElution solid-phase extraction plate, which provided good recovery for all analytes (>92%) and low matrix effects (absolute matrix effects <10%). Standard curves were linear over the range 0.02–250ng/mL with r2 values>0.994. The method was evaluated against current FDA guidelines and was applied to the analysis of patient samples, including an assessment of incurred sample reanalysis (ISR).