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HPLC analysis as a tool for assessing targeted liposome composition.

Mira Oswald1, Michael Platscher2, Simon Geissler1

  • 1Merck Serono, Chemical & Pharmaceutical Development, Frankfurter Straße 250, 64293 Darmstadt, Germany.

International Journal of Pharmaceutics
|November 17, 2015
PubMed
Summary

A new RP-HPLC method enables precise quality control for functionalized phospholipids, essential for developing targeted liposomes. This rapid analysis ensures accurate quantification and identification of components during liposome preparation.

Keywords:
DSPE-PEG(2000)ELSDHPLCLiposomesPhospholipid analysis

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Area of Science:

  • Pharmaceutical Sciences
  • Analytical Chemistry
  • Materials Science

Background:

  • Functionalized phospholipids are critical for designing targeted liposomes.
  • Controlling phospholipid quality and quantity is vital for liposome formulation development and manufacturing.
  • A complex liposomal preparation involves 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), Cholesterol (CHO), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000).

Purpose of the Study:

  • To develop and validate a robust RP-HPLC method for analyzing liposomal components.
  • To ensure accurate quantification and quality control of phospholipids during liposome preparation.
  • To identify potential degradation products within the liposomal formulation.

Main Methods:

  • Development of a reversed-phase high-performance liquid chromatography (RP-HPLC) method.
  • Detection of liposomal components using evaporative light scattering (ELS).
  • Validation of the method for linearity, precision, accuracy, sensitivity, and robustness.

Main Results:

  • The RP-HPLC method demonstrated a non-linear quadratic response for liposomal compounds (0.012-0.42 mg/ml) with R² > 0.99.
  • Method accuracy was confirmed to be 95-105% of the theoretical concentration.
  • Degradation products within the liposomal formulation were successfully identified.

Conclusions:

  • The developed RP-HPLC method serves as an effective control tool for functionalized liposome preparation.
  • HPLC analysis provides a rapid and easy method for monitoring functionalized lipids at each preparation step.
  • This analytical approach is beneficial for the quantification of all components in liposomal formulations.