Selective and Sensitive Method for Estimation of Liraglutide in Human Plasma using Shimadzu LCMS-8060

Significance of the Topic

This study addresses the critical need for accurate quantification of liraglutide, a GLP-1 receptor agonist widely used in type 2 diabetes and obesity management. Precise plasma measurements are essential for pharmacokinetic profiling, dose optimization, and regulatory compliance in drug development and therapeutic monitoring.

Objectives and Study Overview

The primary objective was to develop and validate a rapid, highly sensitive, and high-throughput bioanalytical method for liraglutide in human plasma. Key targets included achieving a lower limit of quantification (LLOQ) of 0.50 ng/mL, minimizing sample volume, and ensuring straightforward method transfer to customer laboratories.

Methodology and Instrumentation

Samples (200 µL plasma) were processed via positive-pressure solid-phase extraction (SPE) with a single-step protocol: conditioning, sample loading, wash, and elution into a methanol-acetonitrile mixture. Chromatographic separation employed a Shim-pack Velox Biphenyl column (100 × 2.1 mm, 2.7 µm) on a Nexera X2 UHPLC system coupled to the Shimadzu LCMS-8060. The mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B), delivered at 0.25 mL/min under a 10-minute gradient. Electrospray ionization in positive MRM mode used a heated interface and UF-Qarray ion guide. The optimized MRM transition for liraglutide was m/z 938.5→1128.4.

Main Results and Discussion

The method exhibited excellent linearity from 0.50 to 202.70 ng/mL (r2 > 0.99). Intra- and inter-day precision and accuracy met US regulatory criteria (≤15% CV and 85–115% accuracy, ≤20%/80–120% at LLOQ). Mean recovery was 50.92% (CV 13.06%), and matrix factor averaged 0.97 (CV 3.69%). No significant carryover or endogenous interferences were observed. Signal-to-noise at the LLOQ exceeded 20:1, demonstrating robust sensitivity.

Benefits and Practical Applications

• High sensitivity (LLOQ 0.50 ng/mL) enables reliable pharmacokinetic studies.
• Low plasma volume (200 µL) reduces sample requirements and prolongs instrument life.
• Single-step SPE simplifies preparation and increases throughput.
• Ready-to-use validated protocol facilitates transfer to routine laboratories.

Future Trends and Applications

Advancements may include automation of SPE workflows, extension to other peptide and protein therapeutics, coupling with high-resolution mass spectrometry for metabolite profiling, and integration of AI-powered data analysis for enhanced throughput and data interpretation. Personalized medicine and therapeutic drug monitoring stand to benefit from further miniaturization and multiplexing.

Conclusion

The validated LCMS-8060 assay provides a selective, sensitive, and high-throughput platform for quantifying liraglutide in human plasma in compliance with major regulatory guidelines. This method supports pharmacokinetic investigations, quality control, and broader analytical applications in clinical and research laboratories.

References

• https://en.wikipedia.org/wiki/Liraglutide (accessed Jan 02, 2020)
• https://www.rxlist.com/victoza-drug.htm (accessed Jan 02, 2020)