Zeta potential: a case study of cationic, anionic, and neutral liposomes

Anal Bioanal Chem. 2017 Sep;409(24):5779-5787. doi: 10.1007/s00216-017-0527-z. Epub 2017 Jul 31.


Zeta potential is often used to approximate a nanoparticle's surface charge, i.e., cationic, anionic, or neutral character, and has become a standard characterization technique to evaluate nanoparticle surfaces. While useful, zeta potential values provide only very general conclusions about surface charge character. Without a thorough understanding of the measurement parameters and limitations of the technique, these values can become meaningless. This case study attempts to explore the sensitivity of zeta potential measurement using specifically formulated cationic, anionic, and neutral liposomes. This study examines zeta potential dependence on pH and ionic strength, resolving power, and highlights the sensitivity of zeta potential to charged liposomes. Liposomes were prepared with cholesterol, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and varying amounts of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS). A strong linear relationship was noted between zeta potential values and the mole percentage of charged lipids within a liposome (e.g., cationic DOTAP or anionic DOPS). This finding could be used to formulate similar liposomes to a specific zeta potential, potentially of importance for systems sensitive to highly charged species. In addition, cationic and anionic liposomes were titrated with up to two mole percent of the neutral lipid 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (lipid-PEG; LP). Very small amounts of the lipid-PEG (<0.2 mol%) were found to impart stability to the DOTAP- and DOPS-containing liposomes without significantly affecting other physicochemical properties of the formulation, providing a simple approach to making stable liposomes with cationic and anionic surface charge.

Keywords: Lipid-PEG; Liposomes; Stability; Surface characterization; Zeta potential.

MeSH terms

  • Anions / chemistry
  • Cations / chemistry
  • Cholesterol / chemistry
  • Fatty Acids, Monounsaturated / chemistry
  • Liposomes / chemistry*
  • Osmolar Concentration
  • Phosphatidylcholines / chemistry
  • Phosphatidylserines / chemistry
  • Polyethylene Glycols / chemistry
  • Quaternary Ammonium Compounds / chemistry
  • Static Electricity
  • Surface Properties


  • Anions
  • Cations
  • Fatty Acids, Monounsaturated
  • Liposomes
  • Phosphatidylcholines
  • Phosphatidylserines
  • Quaternary Ammonium Compounds
  • Polyethylene Glycols
  • 1,2-dioleoylphosphatidylserine
  • Cholesterol
  • 1,2-distearoyllecithin
  • polyethylene glycol 2000
  • 1,2-dioleoyloxy-3-(trimethylammonium)propane