Fully automated platform for determination of Tricyclic Antidepressant in serum

Significance of the Topic

The accurate determination of tricyclic antidepressants (TCAs) in serum is critical for therapeutic drug monitoring due to their narrow therapeutic windows and potential for severe adverse effects. Reliable quantification techniques support clinical decisions in depression, anxiety and pain management, enhancing patient safety and treatment efficacy.

Objectives and Study Overview

This work aimed to develop and validate a fully automated platform for the quantitation of over 13 TCAs in human serum. Key goals included eliminating manual sample preparation, achieving high throughput, and maintaining analytical performance suitable for routine research use.

Methodology and Used Instrumentation

The study employed an online integration of a CLAM-2000 fully automated sample preparation unit with a Nexera X2 HPLC system coupled to a Shimadzu LCMS-8060 triple quadrupole mass spectrometer. Protein precipitation, internal standard spiking and filtration were performed within CLAM-2000 at 8°C, followed by chromatographic separation on a Recipe MS9030 column (40°C) using a binary gradient over 3.8 minutes at 0.6 mL/min. Detection utilized positive‐mode ESI and optimized MRM transitions for each analyte with internal standard calibration.

Main Results and Discussion

• Automation achieved continuous processing and transfer to LC-MS/MS, delivering one quantitative result every five minutes without manual intervention.
• Linearity (R²>0.999) and accuracy (85–115%) were confirmed across low, medium and high concentration calibrators.
• Intra- and inter-day precision (CV%) for all TCAs remained within acceptable analytical ranges (<15%), demonstrating method robustness.
• Chromatographic resolution and sensitivity enabled clear detection down to low-limit of quantification levels, supporting reliable TDM applications.

Benefits and Practical Applications

The fully automated workflow offers multiple advantages:

• Elimination of operator-dependent variability and manual pipetting errors.
• Increased sample throughput and streamlined laboratory operations.
• Reduced reagent consumption and overall cost per analysis.
• Enhanced traceability through software‐controlled procedures and integrated data reporting.

This platform is particularly suited for research environments focused on pharmacokinetic studies, method development and high-throughput screening.

Future Trends and Possibilities

Emerging directions include:

• Extension to additional psychoactive and therapeutic drug classes using expanded MRM panels.
• Integration with laboratory information management systems (LIMS) for end-to-end data handling.
• Miniaturization and multiplexing for simultaneous analysis of larger analyte sets.
• Application of advanced ionization techniques and high-resolution mass spectrometry for enhanced selectivity.

Such advancements will further improve automation, data quality and the scope of clinical and forensic applications.

Conclusion

The presented automated platform successfully integrates sample preparation and LC-MS/MS analysis for rapid, accurate quantification of TCAs in serum. By removing manual steps, the system enhances laboratory efficiency, reduces human error and delivers reliable performance for therapeutic drug monitoring and research workflows.

References

ASMS 2017 ThP090, Vecchietti et al. “Fully automated platform for determination of Tricyclic Antidepressant in serum.” Shimadzu Corporation, June 2017.