BOX-BEHNKEN DESIGN-BASED DEVELOPMENT OF ECO-FRIENDLY RP-HPLC ANALYTICAL METHOD FOR THE QUANTIFICATION OF ERASTIN FROM THE MESOPOROUS SILICA NANOPARTICLES

ASHUTOSH GUPTA; RACHANA S. P.; SUDHEER MOORKOTH; NAMDEV DHAS

doi:10.22159/ijap.2025v17i6.53771

Authors

ASHUTOSH GUPTA Department of Pharmaceutical Quality Assurance, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, Karnataka, India https://orcid.org/0000-0003-4236-6580
RACHANA S. P. Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, Karnataka, India
SUDHEER MOORKOTH Department of Pharmaceutical Quality Assurance, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, Karnataka, India https://orcid.org/0000-0002-5341-0981
NAMDEV DHAS Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, Karnataka, India https://orcid.org/0000-0002-9852-2653

DOI:

https://doi.org/10.22159/ijap.2025v17i6.53771

Keywords:

Box-behnken design, Mesoporous silica nanoparticles, HPLC, Design of expert, Nanoformulation, Greenness

Abstract

Objective: Globally, cancer remains a major source of illness and mortality. Approximately 30% of initial glioblastomas and 80% of all malignant ones are gliomas. Erastin (ERT) is a small molecule that kills cancer cells through ferroptosis and treats brain cancer. The current work aims to optimize, establish, and validate a High-Performance Liquid Chromatography (HPLC) method with sufficient sensitivity and specificity to measure ERT in Mesoporous Silica Nanoparticles (MSNs). The HPLC method was optimized using the Design of Experiments (DoE) technique. The novelty of this project is the development of the DoE-based HPLC method for quantifying ERT from the MSNs.

Methods: The Box-Behnken Design (BBD) was used for the method optimization, 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 conditions were: buffer ratio of 80%, buffer pH 3.8, flow rate 0.9 ml/min, and injection volume 8 µl**. The established HPLC method was validated as per International Council for Harmonisation (ICH) Q2 (R1) recommendations.

Results: The optimization of the HPLC method was done using BBD, and the desirability value was found to be 0.992. The established method showed good linearity (R2 = 0.9991) with a 12-500 ng/ml range, and LOD and LOQ were 2.71 ng and 6.25 ng, respectively. The Mesoporous Silica Nanoparticle (MSNs) matrix did not affect the specificity of our validated HPLC method for the analysis of ERT. Analysing the formulation's surface morphology helped confirm the synthesized MSNs. The proposed HPLC method was effectively used to calculate the medication loaded in the MSNs and to measure the amount of ERT entrapped. The % entrapment efficiency (EE) for ERT in MSNs was found to be 76.07±2.56%, and the % drug loading (DL) for ERT in MSNs was found to be 14±1.87%. The established HPLC method for estimating ERT was found to be environmentally friendly with an overall analytical greenness (AGREE) score of 0.6.

Conclusion: The established HPLC method was developed, optimized, and validated as per ICH Q2 R1 guidelines. The HPLC method was found to be eco-friendly as per the AGREE analysis.

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