Nanoliposomes are extensively used as ideal vehicles in drug delivery systems due to their unique biocompatibility and biodegradability properties. They can be used as sustained release and target selective conveyances to deliver the encapsulated drugs at specific cells or tissues by improving their efficacy along with reducing the side effects. As an analytical perspective, the determination of various lipid components in the final formulation is one of the practical issues while the agents are applied in an industrial-scale. Herein, the maximum ultra violet (UV) absorbances for the most of the lipids are within 200-210 nm that cause significant cut-off conflicts with the general solvents or additives of high-performance liquid chromatography (HPLC) during its method development procedure. In this study, a simple, accurate and cost-effective isocratic HPLC-UV method has been successfully developed for the simultaneous determination of α-(3-O-cholesteryloxy)-δ-(N-ethylmorpholine)-succineamide (MoChol), cholesteryl-hemisuccinate (Chems) and Cholesterol in nanoliposomes drug carriers containing an active pharmaceutical ingredient (anti-BCL-2 DNA oligonucleotide). The isocratic mobile phase consisted of ethanol/acetonitrile/water including trifluoroacetic acid (60/30/10 with 0.1% v/v, respectively) at a flow rate of 1.0 mL min-1 was run through a commercial reverse-phase C18 analytical column while UV detector was set at 202 nm. To confirm the applicability, a full validation of the proposed method was performed according to the International Council for Harmonization (ICH) guidelines.
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