A Simple LC-MS/MS Method for Simultaneous Analysis of 35 Anti-psychotics in Human Plasma for Clinical Research

Significance of the topic

The therapeutic monitoring of antipsychotic drugs in plasma supports personalized dosing, prevents adverse effects, and enhances understanding of pharmacokinetic interactions in clinical research.

Study objectives and overview

This study aimed to establish a rapid, reliable, and cost-effective LC-MS/MS method for simultaneous quantification of 35 antipsychotics in human plasma. Key goals included minimal sample volume, efficient sample preparation, high throughput, and robust performance meeting clinical research requirements.

Methodology

Sample preparation involved protein precipitation:

• 50 µL human plasma spiked with calibrators and quality controls.
• Addition of 100 µL internal standard solution (MeOH:0.1 M ZnSO4, 70:30 v/v), vortex and centrifugation.
• Transfer 100 µL supernatant to 96-well plate and dilute with 100 µL water; mix before injection.

Chromatographic separation used a reversed-phase column with water and methanol mobile phases (2 mM ammonium acetate, 0.1% formic acid) under a gradient flow, achieving a 4.3-minute runtime.

Used instrumentation

• Waters ACQUITY UPLC I-Class system with FL Sample Manager.
• Waters XSelect HSS C18 SB XP column (100 Å, 2.5 µm, 2.1 × 30 mm).
• Waters Xevo TQ-S micro mass spectrometer in positive electrospray ionization mode using MRM.

Main results and discussion

Linearity and sensitivity:

• Excellent linearity (r² > 0.99) across all analytes with 1/x weighting.
• Lower limit of measuring interval achieved at lowest calibrator (≤20% CV, ≤15% bias).
• No significant carryover observed (<25% of the lowest calibrator signal).

Matrix effects and recovery:

• Normalized matrix factors ranged from 0.88 to 1.14, indicating effective compensation by internal standards.
• Extraction recoveries between 85% and 115%; individual analyte extraction efficiencies varied from 52.2% to 96.9%.

Precision and interference:

• Repeatability and total precision ≤8.7% RSD across low, mid, and high QC levels (n = 25).
• No significant interference from common endogenous compounds (albumin, bilirubin, creatinine, lipids, uric acid).

Benefits and practical applications

The method supports high-throughput clinical studies by combining:

• Minimal sample volume (50 µL).
• Fast analysis (<5 minutes per injection).
• Robust quantitative performance suitable for pharmacokinetic and drug interaction studies.

Future trends and potential applications

• Integration with automated sample handling to increase laboratory throughput.
• Expansion of analyte panels to include metabolites and novel antipsychotic agents.
• Application in therapeutic drug monitoring and personalized medicine frameworks.

Conclusion

A straightforward LC-MS/MS protocol enables reliable simultaneous quantification of 35 antipsychotics in human plasma with rapid analysis, strong reproducibility, and minimal matrix interference, facilitating diverse clinical research applications.

References

No literature references were provided in the original document.