A Simple, Accurate, and Cost-effective HPLC-UV for Determination of Nanoliposome Components as Drug Carriers

Background: Nanoliposomes are widely used as drug delivery systems owing to their excellent biocompatibility, biodegradability, and ability to enhance drug stability and targeted delivery. Accurate determination of lipid components in nanoliposomal formulations is essential for quality control, stability assessment, and regulatory compliance; however, simultaneous analysis of structurally similar lipids remains analytically challenging. Objectives: This study aimed to develop and validate a simple, accurate, and cost-effective high-performance liquid chromatography coupled with ultraviolet detection (HPLC–UV) method for the simultaneous determination of α-(۳-O-cholesteryloxy)-δ-(N-ethylmorpholine)-succineamide (MoChol), cholesteryl hemisuccinate, cholesterol, dioleoyl-phosphatidylethanolamine (DOPE) and palmitoyl-oleoylphosphatidylcholine (POPC) in nanoliposome formulations containing anti-BCL-۲ DNA oligonucleotides. Methods: Chromatographic separation was performed on a C۱۸ column using a gradient elution system consisting of ۰.۱% trifluoroacetic acid (TFA) in water and ۰.۱% TFA in ethanol as mobile phases. Detection was performed at ۲۰۲ nm with a flow rate of ۱.۰ mL min⁻¹. The method was validated according to International Council for Harmonization (ICH) guidelines for linearity, accuracy, precision, specificity, robustness, limit of detection (LOD), and limit of quantification (LOQ). Results: Excellent linearity was obtained for all analytes over concentration ranges of ۱–۲۱۳۴ mg L⁻¹ with correlation coefficients (R²) greater than ۰.۹۹۹۷. The LOD and LOQ values ranged from ۰.۱۳–۰.۲۳ mg L⁻¹ and ۰.۳۹–۰.۶۹ mg L⁻¹, respectively. Recovery values were within ۹۸–۱۰۲%, while precision expressed as relative standard deviation was below ۱%. Resolution values exceeded ۲.۰ for all critical peak pairs, demonstrating adequate specificity. The method was successfully applied to evaluate lipid composition in nanoliposome samples and to monitor compositional changes after long-term storage Conclusion: The developed HPLC–UV method provides a reliable, sensitive, and economical approach for the simultaneous determination of major lipid components in nanoliposomal drug delivery systems. Its satisfactory validation performance and applicability to stability studies make it suitable for routine quality control and pharmaceutical development of lipid-based formulations.

کلیدواژه ها:

Nanoliposomes ، Drug carrier ، Lipid analysis ، High-performance liquid chromatography (HPLC) ، Method development