Validation of High-Performance Liquid Chromatography method for the determination of doxorubicin in proliposomal drug delivery system formulation

Abstract

The objective of this study develop a novel proliposome formulation containing Doxorubicin (Dox) and was to validate sensitive and selective reversed-phase high-performance liquid chromatographic (HPLC) method for the evaluation of Dox concentrations of proliposome formulation. The samples were chromatographed on C18 column (Zorbax Eclipse Plus 5µm 4.6 x 250 mm) using a mobile phase with Sodium Lauryl Sulphate solution:Acetonitrile (%50:%50) and detected 254 nm. Linearity was confirmed in the concentration range 10.0–75.0 µg/mL. Specificity, linearity, working range, LOD, LOQ, accuracy, precision, robustness and system suitability studies were done from HPLC validation parameters. Liposome formulation containing Dox was developed by pH gradient method then proliposome formulation was developed with lyophilisation technique. The developed HPLC method, the encapsulation capacity (EE%) was found to be 90%±0.5 and the drug loading capacity (DL%) was found to be 100.0%±0.3. In addition, in vitro release studies and stability study results were evaluated with validated HPLC method. It was observed that developed Dox-proliposome formulation increased Dox release at pH 5.5, pH 6.5 and pH 7.5 by 23.9%, 30.2% and 14.8%, respectively, compared to commercial product. The result of F2 test performed in pH 7.5 media was 51.4%. According to the results of the physicochemical tests performed within the stability studies, it was observed that there was no significant change at the end of 12 months. These results show that the HPLC method developed, and validation study performed are important and applicable in the development, characterization, in vitro release and stability studies of the novel proliposome formulation.