Highly sensitive LC-MS/MS method for the quantification of barnidipine in human plasma, using the SCIEX QTRAP® 6500+ System

Importance of Bioanalytical Barnidipine Quantification

Barnidipine, a long-acting calcium channel blocker in the dihydropyridine class, exhibits very low plasma concentrations during pharmacokinetic studies. Accurate quantification at picogram-per-milliliter levels is essential for dose optimization, safety evaluation, and support of drug development efforts.

Study Objectives and Overview

This work aimed to develop and validate a highly sensitive LC-MS/MS assay on the SCIEX QTRAP 6500+ system for measuring barnidipine in human plasma. The method targets a calibration range of 5–8005.88 pg/mL, aligned with regulatory guidelines, and seeks to establish a low limit of detection for trace-level bioanalysis.

Methodology and Instrumentation

• Sample Preparation: Human plasma samples were spiked with barnidipine calibration standards and quality controls.
• Chromatography: Reversed-phase liquid chromatography separated analytes under optimized gradient conditions.
• Mass Spectrometry: The SCIEX QTRAP 6500+ system, equipped with electrospray ionization, operated in multiple reaction monitoring (MRM) mode to enhance sensitivity and selectivity.
• Calibration and Sensitivity: Calibration curve spanned 5 to 8005.88 pg/mL; limit of detection (LOD) in neat solution was determined at 0.1 pg/mL.

Main Results and Discussion

• Linearity: Demonstrated excellent linear response across the calibration range with correlation coefficients meeting regulatory acceptance criteria.
• Sensitivity: Achieved an LOD of 0.1 pg/mL, enabling reliable detection of barnidipine at trace concentration levels in plasma.
• Precision and Accuracy: Quality control samples showed intra- and inter-run variation within ±15% of nominal values, satisfying bioanalytical method validation standards.
• Chromatographic Performance: Representative chromatograms confirmed clear peak resolution and minimal endogenous interference.

Benefits and Practical Applications

This validated bioanalytical method supports detailed pharmacokinetic profiling of barnidipine in clinical and preclinical studies, facilitates informed dose selection, and underpins safety monitoring. Its high sensitivity and robustness make it well suited for routine use in pharmaceutical research and quality control laboratories.

Future Trends and Potential Applications

• Advancements in Ionization Techniques: Adoption of novel ion sources may further enhance detection limits.
• Microflow and Nano-LC Integration: Lower solvent consumption while improving sensitivity.
• Automation and High-Throughput Workflows: Streamlined sample preparation and data processing for large-scale studies.
• Personalized Medicine: Application to individualized therapeutic drug monitoring and pharmacokinetic modeling.

Conclusion

The SCIEX QTRAP 6500+-based LC-MS/MS method provides a highly sensitive, reproducible, and regulatory-compliant approach for quantifying barnidipine in human plasma at picogram levels. It addresses critical bioanalytical challenges and supports robust drug development programs.

References

1. DH Tech. Dev. Pte. Ltd. Application Note RUO-MKT-07-10303-A, 2019.