Fast-Track Quantitative Analysis of Anti-Arrhythmic Drugs Using a Laser Diode Thermal Desorption (LDTD) coupled to a Mass Spectrometer

Fast-Track Quantitative Analysis of Anti-Arrhythmic Drugs Using a Laser Diode Thermal Desorption (LDTD) coupled to a Mass Spectrometer

Authors: Anastasiia Shyian¹, Anna Lenartowicz², Maciej Stopa¹, Katarzyna Krupczyńska-Stopa², Rafał Szewczyk^1,2, Serge Auger ³

1. 1. LabExperts sp. z o.o., 7A Limbowa Str., 80-175 Gdansk, Poland
   2. Bioanalytic sp. z o. o., 7A Limbowa Str., 80-175 Gdansk, Poland
   3. Phytronix LLC, Québec, Canada

Introduction

Cardiac arrhythmia affects approximately 12.6% of people over the age of 65 and represents a major clinical challenge. COVID-19 infection is associated with mid- and long-term accelerated vascular ageing, especially in women, and may contribute to the development or progression of cardiovascular disease, increasing arrhythmia burden. Pharmacological therapy requires maintaining optimal drug concentrations for safety and efficacy. To support therapeutic drug monitoring, a rapid method for simultaneous analysis of multiple anti-arrhythmic drugs and a metabolite was developed using laser diode thermal desorption coupled to mass spectrometry. The approach eliminates the need for HPLC and significantly reduces analysis time. In the Axino system, samples on Domino LazzWell plates are rapidly heated by an infrared laser for fast analyte desorption efficiency.

Methods

Serum samples were prepared using a validated protocol. Saturated sodium chloride solution was added, followed by extraction with acetonitrile. The organic upper phase was evaporated under a nitrogen stream, and the residue was reconstituted in a dedicated reconstitution solution to facilitate efficient desorption of analytes from the Domino LazzWell. For analysis, 4 µL of sample was deposited onto the plate. Samples were then analyzed using . Detection was performed in positive ionization mode using multiple reaction monitoring (MRM) on a QTRAP 6500+ mass spectrometer (SCIEX). The method enables simultaneous quantitation of four cardiovascular drugs: metoprolol, bisoprolol, propafenone, and eplerenone, as well as the pharmacologically active metabolite of amiodarone: desethylamiodarone.

Preliminary results

A rapid analytical method for simultaneous quantitation of selected anti-arrhythmic drugs and one metabolite was successfully developed using laser diode thermal desorption coupled to mass spectrometry (LDTD-MS). This approach eliminates the need for HPLC separation, removing the requirement to buy or prepare mobile phases and to equilibrate chromatography columns, significantly simplifying the analytical workflow. Direct analysis by LDTD-MS reduced the total analysis time to approximately 6 seconds per sample, enabling high-throughput measurements suitable for therapeutic drug monitoring applications. A validated sample preparation and analysis procedure was established. Validation demonstrated linearity for all analytes (r ≥ 0.995), accuracy between 80–120%, and reproducibility with %CV ≤ 15%. The method covers the validated concentration ranges for the analyzed compounds: 2.5-250 ng/mL for metoprolol, bisoprolol, 10-250 ng/ml propafenone, and 25-2500 ng/mL for eplerenone and desethylamiodarone; these ranges cover the therapeutic concentrations of all tested drugs.

Overall, the method demonstrates the feasibility of rapid, multiplexed analysis of cardiovascular drugs and their metabolite without the limitations of conventional LC-MS/MS workflows. By eliminating HPLC requirements and reducing analysis time to seconds per sample, this LDTD-MS approach provides a highly efficient and low-cost tool for therapeutic drug monitoring in clinical and research applications, while maintaining accuracy, linearity, and reproducibility across the therapeutic ranges of all compounds.

Novel aspect

HPLC-free LDTD–MS enables rapid, high-throughput therapeutic drug monitoring of multiple anti-arrhythmic drugs.

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