Therapeutic drug monitoring of Antiepileptic drugs in serum: a fully automated approach

Significance of the Topic

The therapeutic drug monitoring (TDM) of antiepileptic drugs (AEDs) is essential for optimizing patient therapy and minimizing adverse effects. Due to high inter- and intra-individual variability in pharmacokinetics, precise and reliable quantification of multiple AEDs in serum is a critical requirement in both research and clinical settings.

Study Objectives and Overview

This study presents a fully automated workflow for simultaneous quantitation of 26 common AEDs in human serum. By coupling an automated sample preparation unit (CLAM-2000) with a triple quadrupole LC-MS/MS system (Nexera X2-LCMS-8060), the aim was to eliminate manual extraction steps, enhance throughput, and maintain high analytical accuracy.

Methodology and Instrumentation

• Sample Preparation: On-deck protein precipitation, filtration, and extract transfer performed by CLAM-2000 at 8 °C.
• Chromatography and Detection: Nexera X2 UHPLC linked to LCMS-8060 triple quadrupole; multiple reaction monitoring (MRM) transitions set for each analyte.
• Calibration: Three serum calibrator levels and internal standards (ClinCal® MS9213 and ClinMass® MS9212) used to evaluate linearity and accuracy.

Key Results and Discussion

• Throughput: One fully quantified sample every 6 minutes without human intervention.
• Correlation: Excellent agreement with manual preparation (Passing-Bablok regression across 20 spiked samples).
• Linearity and Accuracy: R2 values ≥ 0.97 for most analytes, accuracy within 85–115%.
• Precision: Intraday and interday CVs below 15% across low and high concentration controls.

Method Benefits and Practical Applications

The automated approach reduces operator risk, reagent consumption, and per-sample costs while improving reproducibility. It is well suited for high-throughput TDM in research and quality control laboratories.

Future Trends and Potential Applications

• Extension to broader drug panels and biomarkers.
• Integration with laboratory information management systems for real-time reporting.
• Miniaturized cartridges and on-site point-of-care platforms.
• Combining TDM data with pharmacogenomic profiling for personalized therapy.

Conclusion

The fully automated LC-MS/MS workflow provides a robust, accurate, and cost-efficient solution for simultaneous monitoring of multiple AEDs in serum, paving the way for streamlined therapeutic monitoring in both research and clinical laboratories.