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.
Highlights
Liposomes continue to be a major focus in drug delivery research due to their ability to enhance the delivery and targeting of a wide range of drugs and vaccines
The adjuvants are based on a two-component vesicle formulation, which is the combination of the cationic lipid dimethyldioctadecylammonium (DDA) and D-(+)-trehalose 6,60 -dibehenate (TDB)
The required amount of lipid solution was transferred to a round-bottom flask to reach the desired lipid concentration and mixed; DDA and TDB concentrations were fixed at 2.5 and 0.5 mg/mL, respectively, and cholesterol was added to the CAF01 formulation at a concentration of 0.8 mg/mL
Summary
Liposomes continue to be a major focus in drug delivery research due to their ability to enhance the delivery and targeting of a wide range of drugs and vaccines. The combination of DDA and TDB within the liposome formulation was shown to further enhance the adjuvant properties of the liposomes [10]. Beam illuminates the sample, and when the laser hits the particles, the intensity of the scattered light is according by to the scattered light detected This method is constrained to a sufficiently volatile solvent measured a detector DDA:TDB liposome adjuvant system and to demonstrate the general procedures such that they does not require an initial lipid extraction process. This method was used to quantify lipids within can be appliedliposome to a wide range of lipidsand used bytoliposomologists. We consider the ability of this HPLC-ELSD method to detect phospholipid lipid degradation
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