Lipid Nanoparticle and Liposome Characterization with FFF-MALS-DLS

Significance of the topic

Lipid-based nanocarriers such as liposomes and lipid nanoparticles offer precise control over drug encapsulation, stability and release kinetics, making them essential tools in pharmaceuticals and therapeutics. Accurate characterization of particle size, shape and structure is critical for ensuring efficacy, safety and regulatory compliance, motivating the adoption of high-resolution analytical methods.

Objectives and study overview

This white paper demonstrates the combined field-flow fractionation with multi-angle and dynamic light scattering (FFF-MALS-DLS) as a robust technique for detailed characterization of lipid nanoparticles and liposomes. It compares FFF-MALS-DLS with common methods (batch DLS, EM, NTA), and presents three case studies: solid lipid nanoparticles and nanostructured lipid carriers, preclinical stability and serum interaction studies, and drug transfer kinetics between liposomal formulations.

Methodology and instrumentation

• Separation: Eclipse asymmetrical FFF system coupled to standard HPLC components (pump, degasser, autosampler).
• Online characterization: DAWN MALS detector for radius of gyration (Rg) and WyattQELS DLS module for hydrodynamic radius (Rh).
• Additional detection: UV/Vis and fluorescence detectors for composition and drug quantification.

Main results and discussion

• Case 1: FFF-MALS revealed elongated structures in solid lipid nanoparticles and nanostructured carriers, confirmed by elevated Rg values and EM imaging, contrasting with spherical nano-emulsions.
• Case 2: Stability and serum incubation studies detected subtle changes in size distribution and particle morphology not observable by batch DLS, demonstrating FFF-MALS-DLS high sensitivity in complex media.
• Case 3: A label-free FFF-MALS-UV/fluorescence assay enabled baseline separation of donor and acceptor liposomes and precise measurement of drug transfer kinetics, dependent on lipid mass ratios.

Benefits and practical applications

FFF-MALS-DLS delivers high-resolution, quantitative size and shape distributions over large particle ensembles, addresses biases of traditional methods, and meets critical quality attribute requirements for regulatory submissions. It supports drug loading assessment, release kinetics, and stability studies in pharmaceutical development and quality control.

Future trends and opportunities

• Integration with preparative FFF channels for higher sample loading.
• Expansion to additional detectors for compositional analysis.
• Development of ex vivo predictive assays for drug release and transfer.
• Broader application to diverse nanocarrier systems and adoption in regulatory frameworks.

Conclusion

The FFF-MALS-DLS platform provides an indispensable analytical approach for comprehensive characterization of lipid-based drug carriers, enabling precise size, shape and compositional analysis to support research, development and regulatory compliance in nanomedicine.

Used instrumentation

• Eclipse asymmetrical field-flow fractionation device (Wyatt Technology).
• DAWN multi-angle light scattering detector.
• WyattQELS dynamic light scattering module.
• HPLC system components: pump, degasser, autosampler.
• UV/Vis and fluorescence detectors.

References

1. Johann C., Waters|Wyatt Technology. Lipid Nanoparticle and Liposome Characterization with FFF-MALS-DLS. White Paper WP2608.
2. Podzimek S. Light Scattering, Size-exclusion Chromatography and AF4. John Wiley & Sons; 2012.
3. Jores K., Mehnert W., Drechsler M., Bunjes H., Johann C., Mäder K. Investigations on solid lipid nanoparticles and nanostructured carriers by photon correlation spectroscopy, field-flow fractionation and TEM. J Control Release. 2004;95(2):217-227.
4. Caputo F., et al. Measuring Particle Size Distribution by AF4: Preclinical Characterization of Lipid-Based Nanoparticles. Mol Pharm. 2019;16(2):756-767.
5. Hinna A.H., Huzfeld S., Kuntsche J., Bauer-Brandl A., Brandl M. Mechanism and kinetics of drug loss from liposomal carriers studied by AF4-based drug release/transfer assay. J Control Release. 2016;232:228-237.
6. FDA. Guidance for Industry: Liposome Drug Products. April 2018.