Journal of pharmaceutical and biomedical analysisJul 1, 2026

How the type, purity, and stability of charged lipids affect lipid nanoparticle performance

mRNA Technology Weekly Brief ↗PubMed ↗DOI ↗

Updated Jul 2, 2026

Abstract

Four structurally different ionizable lipids were evaluated, revealing significant variability in transfection efficiency based on lipid structure and cell type.

The performance of messenger RNA (mRNA)-loaded lipid nanoparticles (LNP) is influenced by the identity, purity, and stability of ionizable lipids.
Batch-to-batch variability and stability of lipids are critical for maintaining in vitro potency of LNP formulations.
Impurities derived from lipid synthesis or degradation can lead to significantly reduced mRNA expression in cell cultures.
No apparent differences in traditional physicochemical characteristics, such as particle size distribution or encapsulation efficiency, were observed despite variations in potency.
Advanced analytical techniques enabled the detection and quantification of impurities, correlating their presence with a loss in transfection potency.

Simplified

Lipid nanoparticles (LNP) have become the leading platform for nucleic acid delivery. However, achieving reproducible and efficacious LNP formulations remains a challenge. Although ionizable lipids are recognized as key determinant for LNP performance, the factors governing efficient RNA delivery are still not fully understood. In this work, we systematically investigated the influence of ionizable lipid identity, purity, and stability on the performance of messenger RNA (mRNA)-loaded LNP. Four structurally different ionizable lipids were evaluated, revealing that transfection efficiency is strongly lipid structure- and cell type-dependent, emphasizing the need of tailoring lipids and, thus, LNP to specific therapeutic applications. Furthermore, batch-to-batch variability and stability studies showed that lipid purity and chemical integrity are critical for LNP's in vitro potency. Lipids containing synthesis- or degradation-derived impurities exhibited markedly reduced mRNA expression in cell culture, despite no apparent differences in "conventional" physicochemical characteristics such as particle size distribution or encapsulation efficiency. Advanced analytical methods, including high performance liquid chromatography with a charged aerosol detector (HPLC-CAD) and liquid chromatography-high resolution mass spectrometry (LC-HRMS), enabled precise detection and quantification of impurities, directly correlating their presence with loss in transfection potency. Overall, this work provides valuable insights into the quality-performance relationships of ionizable lipids utilized in LNP formulations. It underscores the importance of rigorous raw material characterization and stability testing to ensure the reliability of LNP delivery technology.

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Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

PubMed

How the type, purity, and stability of charged lipids affect lipid nanoparticle performance

Abstract

Full Text

Funding

Competing interests

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