Quality by design based development and validation of HPLC method for simultaneous estimation of paclitaxel and vinorelbine tartrate in dual drug loaded liposomes

Abstract

This paper describes the development of HPLC method for simultaneous estimation of paclitaxel and vinorelbine tartrate loaded in dual drug liposomes, using quality by design (QbD) approach. The main objective was to identify the robust chromatographic conditions where an adequate separation of the components with quality peaks, within acceptable run time can be achieved. Based on this objective, Target analytical profile (TAP) was defined and systematic risk analysis was carried out to identify critical method attributes (CMA) having impact on critical quality attributes (CQA). % Organic phase, pH, and ammonium acetate concentration in the aqueous phase were identified as CMA. Box-Behnken design was employed to establish the quantitative relationship between CMA and CQA which was further utilized to generate analytical design space and to develop a control strategy. The effective chromatographic separation was accomplished using C18 (4.6 mm, 100 mm, dp 5 μ) column, and mobile phase consisting of acetonitrile – aqueous phase (30 mM ammonium acetate adjusted to pH 3.2 using ortho phosphoric acid) (55:45, v/v) at ambient temperature and at flow rate of 1 mL/min. The elution was monitored at 249 nm using an ultraviolet detector. This developed HPLC method was validated using ICH guidelines. The method has been successfully used for quality analysis of development batches of dual drug liposomes and stability samples and will be applicable throughout the life cycle of the product.