Rapid Analysis of Lipid Nanoparticle Components Using BioAccord™ LC-MS System

Significance of the Topic

The successful deployment of mRNA vaccines against SARS-CoV-2 has highlighted the critical role of lipid nanoparticle (LNP) delivery systems. Encapsulation of mRNA in LNP protects it from enzymatic degradation, enhances cellular uptake, and improves protein expression efficiency.

Study Objectives and Overview

This study aimed to develop a straightforward, rapid, and routine reversed-phase LC-MS method for comprehensive characterization of the four main LNP lipids—cholesterol, phospholipid DSPC, ionizable lipid MC3, and PEGylated lipid DMG-PEG-2000—using the ACQUITY Premier CSH C18 column coupled to the BioAccord LC-MS system.

Methodology and Instrumentation

• LC Conditions: Mobile phase A comprised acetonitrile/water/1 M ammonium formate (600/390/10, 0.1 % formic acid); phase B was isopropanol/acetonitrile/1 M ammonium formate (900/90/10).
• Column: ACQUITY Premier CSH C18 (100 × 2.1 mm).
• MS Conditions: Positive electrospray ionization, m/z range 50–2000, cone voltage 30 V, fragmentation voltage ramp 120–200 V.
• Instrument: BioAccord LC-MS system.

Key Results and Discussion

• Single Lipid Detection: Distinct extracted ion chromatograms and spectra were obtained for each LNP component, confirming effective separation of spectroscopically silent species.
• Complex Lipid Analysis: The PEGylated lipid displayed multiple charge states (+2 to +4) and variable PEG chain lengths (38–50 repeat units), demonstrating method sensitivity.
• New Peak Detection: Binary comparison of reference and spiked samples enabled rapid identification of additional lipid impurities through difference chromatograms and matched fragment ions.

Benefits and Practical Applications of the Method

The developed workflow provides high-throughput, routine analysis of LNP formulations, supporting identification of individual components, degradation products, process and raw material impurities, and quality control during vaccine development and manufacturing.

Future Trends and Potential Applications

• Adoption of high-resolution MS/MS for deeper structural elucidation of novel lipid variants.
• Automation and machine-learning–driven data analysis for accelerated impurity profiling.
• Extension to next-generation LNP platforms and personalized therapeutic formulations.
• Implementation of scalable QC workflows for continuous manufacturing of mRNA products.

Conclusion

A streamlined reversed-phase LC-MS method using the BioAccord system and ACQUITY Premier CSH C18 column enables rapid, reliable analysis of key LNP components, offering valuable support for vaccine formulation development and quality assurance processes.

References

1. Vaccines 2021, 9(1), 65; DOI:10.3390/vaccines9010065