LC-MS/MS Method Development and Analysis of Tricyclic Antidepressants in Human Plasma

Author

Shahbaz GulFollow

Date of Award

Spring 5-2021

Document Type

Undergraduate Thesis

Department

Pharmacy Administration

First Advisor

Waseem Gul

Second Advisor

Mohamed M. Radwan

Third Advisor

Suman Chandra

Relational Format

Dissertation/Thesis

Abstract

Depression has become one of the most prominent problems in society, disrupting both personal and social lives. Tricyclic antidepressants (TCAs) were first developed in the 1950’s and became some of the leading anti-depressant medications on the market later that decade. Today, newer anti-depressants have risen to the forefront, being safer and having a lower side-effect probability. Nonetheless, TCAs continue to be prescribed for severe depression, especially in cases where the newer anti-depressants have failed. However, TCAs are still highly potent, the toxicity associated with these compounds cannot be ignored. They have considerable cardiovascular and neurological toxicity, and in the event of an overdose, may lead to death within an hour. In an effort to identify the concentration of these TCAs in individuals, an LC-MS/MS method was developed and validated for the analysis of amitriptyline, desipramine, imipramine, and nortriptyline in human plasma samples. The method was developed and validated on two different LC-MS/MS instruments, ABSciex QTRAP 3200 and ABSciex QTRAP 4500. Two specific MRM transitions for amitriptyline, desipramine, imipramine, and nortriptyline, and two specific MRM transitions for amitriptyline-d₃, desipramine-d₃, imipramine-d₃, and nortriptyline-d₃ internal standards, were monitored for each compound for maximum selectivity and sensitivity. Separation was achieved on a Synergi Hydro-RP column (150 x 3.00 mm; 4 µm; 80 Å), and data acquisition and processing were performed with Analyst^TM 1.6.3 software. The extraction procedure was developed to be rapid and simple, involving crashing the plasma with cold acetonitrile (ACN), filtering through a 0.2 μ polypropylene (PP) filter, evaporation, and reconstitution. The limit of detection (LOD) and limit of quantitation (LOQ) were identified to be 1 ng/mL, and the upper limit of linearity (uLOL) was determined to be 400 ng/mL. The developed method was applied to more than 350 human plasma samples. Matrix effect and drug interference studies were carried out, and neither matric effect nor drug interference was observed in any of the samples. The developed and validated LC-MS/MS method was robust, reproducible, and highly sensitive for the determination of low levels of the TCAs in human plasma samples.

Recommended Citation

Gul, Shahbaz, "LC-MS/MS Method Development and Analysis of Tricyclic Antidepressants in Human Plasma" (2021). Honors Theses. 1598.
https://egrove.olemiss.edu/hon_thesis/1598

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