Rapid Quantification and Validation of Lipid Concentrations within Liposomes

Carla Roces, Elizabeth Kastner, Peter Stone, Deborah Lowry, Yvonne Perrie

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)
137 Downloads (Pure)


Quantification of the lipid content in liposomal adjuvants for subunit vaccine formulation is of extreme importance, since this concentration impacts both efficacy and stability. In this paper, we outline a high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method that allows for the rapid and simultaneous quantification of lipid concentrations within liposomal systems prepared by three liposomal manufacturing techniques (lipid film hydration, high shear mixing, and microfluidics). The ELSD system was used to quantify four lipids: 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), cholesterol, dimethyldioctadecylammonium (DDA) bromide, and D-(+)-trehalose 6,6′-dibehenate (TDB). The developed method offers rapidity, high sensitivity, direct linearity, and a good consistency on the responses (R2 > 0.993 for the four lipids tested). The corresponding limit of detection (LOD) and limit of quantification (LOQ) were 0.11 and 0.36 mg/mL (DMPC), 0.02 and 0.80 mg/mL (cholesterol), 0.06 and 0.20 mg/mL (DDA), and 0.05 and 0.16 mg/mL (TDB), respectively. HPLC-ELSD was shown to be a rapid and effective method for the quantification of lipids within liposome formulations without the need for lipid extraction processes.
Original languageEnglish
Pages (from-to)1-11
Issue number29
Early online date13 Sept 2016
Publication statusPublished online - 13 Sept 2016


  • lipids
  • liposomes
  • cholesterol
  • quantification
  • HPLC
  • ELSD


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