Simultaneous estimation of neratinib and naringenin from pharmaceuticals using a stability-indicating liquid chromatographic method: Application to analytical quality by design concept and multi-tool method greenness assessment

A validated stability-indicating simultaneous liquid chromatographic method was developed to estimate neratinib (NRB) and naringenin (NRN). We used the principles of analytical quality by design (AQbD) to develop a robust analytical method. The ACN%, pH, and flow rate were the three method variables investigated and optimized using the Box-Behnken experimental design (BBD). The optimized chromatographic condition used a mobile phase of ACN: phosphate buffer pH 3.8 (70:30 v/v) flowing at 0.6 mL.min⁻¹ on a 250 mm C-18 column. Diode array detection was performed at 277 nm. The method was robust, linear (r² = 0.999), accurate and precise. The detection and quantitation limits were 0.275 and 1.0 µg.mL⁻¹ for both drugs. Forced degradation revealed non-interference of degradation peaks with the analyte peaks and ensured method specificity. The LC estimation demonstrated higher solubility of NRB and NRN in Precirol ATO5 and Tween 80 as solid lipid and surfactant, respectively. In addition, the entrapment efficiencies were 78 % and 86 % for SLNs of NRB and NRN, respectively. After 24 h, the In-vitro release study indicated that 40 % of the drugs were released from the pure drug suspension and >95 % released from the SLN. Afterwards, the results from five different greenness assessment approach established the present method's green nature. Overall, a green, federally robust, and flexible liquid chromatographic method was developed and can be applied routinely to estimate NRB and NRN from the developed nano formulation.