Ion quantification in liposomal drug products using high performance liquid chromatography

J Pharm Biomed Anal. 2019 Feb 20;165:41-46. doi: 10.1016/j.jpba.2018.11.048. Epub 2018 Nov 22.


A simple, straightforward analytical method based on liquid chromatography has been optimized to quantify total, internal, and external ions in drug-loaded liposomal products. The quantification of ammonium and sulfate ions in Doxil is detailed; although, the methodology has been extrapolated to quantitate a variety of ions, including calcium, acetate, and others in several different liposomal formulations. Total ion concentrations were measured after disruption of the liposome via lyophilization, to liberate all components. External ion concentrations were made following membrane centrifugation, without disruption of the liposome structure, where the permeate fraction was analyzed for external ion quantities. The internal ion fraction was derived from mass balance of the total and external ion measurements. High performance liquid chromatography (HPLC), equipped with different separation columns, and coupled to a charged aerosol detector, was employed for all ion quantifications. The analytical measurements were confirmed using simple stoichiometry based on the drug crystallization of doxorubicin within the liposome interior. The method presented herein is quick, highly accurate, and has significantly improved lower limits of detection and quantification over other traditional methods. As more follow-on versions of Doxil are being developed, this facile approach to ion quantitation can be used to help establish compositional similarity to the reference listed drug.

Keywords: Ammonium; Doxil; Ion concentration; Liposome; Nanoparticle; Sulfate.

MeSH terms

  • Antibiotics, Antineoplastic / analysis*
  • Antibiotics, Antineoplastic / chemistry
  • Chromatography, High Pressure Liquid / methods*
  • Crystallization
  • Doxorubicin / analogs & derivatives*
  • Doxorubicin / analysis
  • Doxorubicin / chemistry
  • Drug Delivery Systems
  • Freeze Drying
  • Ions
  • Limit of Detection
  • Liposomes
  • Polyethylene Glycols / analysis
  • Polyethylene Glycols / chemistry
  • Reproducibility of Results


  • Antibiotics, Antineoplastic
  • Ions
  • Liposomes
  • liposomal doxorubicin
  • Polyethylene Glycols
  • Doxorubicin