Rapid quantification and validation of lipid concentrations within liposomes

Carla B. Roces, Elisabeth Kastner, Peter Stone, Deborah Lowry, Yvonne Perrie*

*Corresponding author for this work

Research output: Contribution to journalSpecial issue

Abstract

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
Article number29
JournalPharmaceutics
Volume8
Issue number3
DOIs
Publication statusPublished - 13 Sep 2016

Fingerprint

Liposomes
Lipids
Trehalose
Phosphorylcholine
Cholesterol
High Pressure Liquid Chromatography
Light
Subunit Vaccines
Microfluidics
Limit of Detection

Bibliographical note

© 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).

Funding: EPSRC (EP/I033270/1); EU Horizon 2020 (643381); and the EPSRC and the BBSRC (Industrial Case Award BB/L017245/1).

Data can be found at DOI: 10.15129/4338bb40-6f84-4dde-b275-6f1656cc5c05.

Keywords

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

Cite this

Roces, Carla B. ; Kastner, Elisabeth ; Stone, Peter ; Lowry, Deborah ; Perrie, Yvonne. / Rapid quantification and validation of lipid concentrations within liposomes. In: Pharmaceutics. 2016 ; Vol. 8, No. 3.
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Rapid quantification and validation of lipid concentrations within liposomes. / Roces, Carla B.; Kastner, Elisabeth; Stone, Peter; Lowry, Deborah; Perrie, Yvonne.

In: Pharmaceutics, Vol. 8, No. 3, 29, 13.09.2016.

Research output: Contribution to journalSpecial issue

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AU - Roces, Carla B.

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AU - Stone, Peter

AU - Lowry, Deborah

AU - Perrie, Yvonne

N1 - © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/). Funding: EPSRC (EP/I033270/1); EU Horizon 2020 (643381); and the EPSRC and the BBSRC (Industrial Case Award BB/L017245/1). Data can be found at DOI: 10.15129/4338bb40-6f84-4dde-b275-6f1656cc5c05.

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AB - 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.

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