Simultaneous Determination of Polyethylene Glycol-Conjugated Liposome Components by Using Reversed-Phase High-Performance Liquid Chromatography with UV and Evaporative Light Scattering Detection

Abstract

Abstract. Liposomes incorporating polyethylene glycol (PEG)-conjugated lipids (PEGylated liposomes) have attracted attention as drug delivery carriers because they show good in vivo stability. The lipid component of PEGylated liposomal formulations needs to be quantified for quality control. In this study, a simple reversed-phase high-performance liquid chromatography (HPLC) method with an evaporative light-scattering detector (ELSD) was established for simultaneous determination of hydrogenated soy phosphatidylcholine, cholesterol, PEG-conjugated lipid, and hydrolysis products of phospholipid in PEGylated liposomal formulations. These lipids were separated using a C18 column with a gradient mobile phase consisting of ammonium acetate buffer and ammonium acetate in methanol at a flow rate of 1.0 ml/min. This method provided sufficient repeatability, linearity, and recovery rate for all lipids. However, the linearity and recovery rates of cholesterol achieved using a ultraviolet (UV) detector were better than those achieved using an ELSD. This validated method can be applied to assess the composition change during the preparation process of liposomes and to quantify lipid components and hydrolysis products contained in a commercially available liposomal formulation DOXIL®. Taken together, this reversed-phase HPLC-UV/ELSD method may be useful for the rapid or routine analysis of liposomal lipid components in process development and quality control.

Determination of Key Parameters for a Mechanism-Based Model to Predict Doxorubicin Release from Actively Loaded Liposomes

“…Lipid analyses were performed on the same HPLC system and column described above but with a SEDEX 85 LT‐ELSD detector (Sedere, Alfortville, France) for lipid quantitation operated at 45°C under 3.5 bar of ultra‐high purity nitrogen gas. A previously published method was adapted in order to perform isocratic separations using methanol containing 4 mM ammonium formate as the mobile phase at a flow rate of 1 mL/min. The sample compartment was at 5°C and the column was thermostated at 45°C.…”

Development and validation of a novel UPLC-ELSD method for the assessment of lipid composition of nanomedicine formulation

“…ELSD has become popular to detect and monitor the separation of poor UV-absorbers, such as phospholipids [40][41][42][43][44], PEG [45][46][47][48] and triglycerides [49][50][51][52]. This detection technique was chosen in these studies for several reasons.…”

An efficient PEGylated liposomal nanocarrier containing cell-penetrating peptide and pH-sensitive hydrazone bond for enhancing tumor-targeted drug delivery

“…16,17 The resolution (R) between PSL (or CL) and PEG exceeded 3.2, representing a satisfactory separation between them ( Figure 3A). At pH 7.0, the PEG peak did not appear in PSL, indicating that PSL was stable and PEG could not be cleaved from the liposomes.…”