Box-Behnken design-based development of eco-friendly RP-HPLC analytical method for the quantification of erastin from the mesoporous silica nanoparticles

Abstract

The current work aims to optimize, stablish, validate a high-performance liquid chromatography (HPLC) method that possesses sufficient sensitivity and specificity to measure erastin (ERT) in mesoporous silica nanoparticles. The HPLC method was optimized using Design of Experiments (DoE) technique. The Box-Behnken design (BBD) was used and buffer ratio, injection volume, buffer pH, and flow rate were used as the independent factors. The dependent variables were retention time (Rt) of ERT, peak area of ERT, and tailing factor (Tf) of ERT. The optimized chromatographic condition were: buffer ration 80%, buffer pH 3.8 and flow rate 0.9 mL/min, and injection volume 8 µL. The established HPLC method was validated as per ICH Q2 (R1) recommendations. The established method showed the good linearity (R2 = 0.9991) having a range of 12–500 ng/mL and LOD and LOQ was 2.71 ng and 6.25 ng, respectively. The mesoporous silica nanoparticle (MSN) matrix did not affect the specificity of our validated RP-HPLC method of analysis for ERT. Analysing the formulation's surface morphology helped with the confirmation of the synthesized MSNs. The proposed RP-HPLC method was effectively used to calculated the medication loaded in the MSNs as well as to measure the amount of ERT entrapped. The % EE for ERT in MSNs was found to be 76.07% and % DL for ERT in MSNs was found to be 14%, respectively. The established HPLC method was found ecofriendly as per the AGREE analysis.