# Reverse Phase High Performance Liquid Chromatography Method for the Estimation of Mangiferin in Nano Formulations: Development and Optimization by Analytical Quality-by-Design

## TL;DR

This paper presents a new HPLC method optimized using QbD for measuring mangiferin in nano-formulations, ensuring reliable and robust analysis.

## Contribution

A novel RP-HPLC method for mangiferin in nano-formulations, developed and optimized using QbD and validated for quality control.

## Key findings

- The method was validated and found to meet ICH Q2(R2) guidelines for accuracy, precision, and robustness.
- The method successfully analyzed the stability and release kinetics of MGF SLNs.
- The developed RP-HPLC method is suitable for routine quality control of nanocarrier systems.

## Abstract

A novel reverse-phase
high-performance liquid chromatography (RP-HPLC)
method was developed and optimized for the estimation of Mangiferin
(MGF) by employing a Quality by Design (QbD) approach. Preliminary
method screening was done by the Taguchi OA, followed by using a Box–Behnken
Design (BBD), enabling systematic evaluation of critical factors with
a reduced number of experimental runs. The model efficiently established
significant correlations between selected variables and analytical
responses, thereby enhancing the method’s robustness and reliability.
Chromatographic separation was achieved on a Hyperclone C18 column
(4.6 × 250 mm, 5 μm) using an isocratic mobile phase consisting
of phosphate buffer (pH 3.2) and acetonitrile (30%) + methanol (70%)
in a 78:22% v/v ratio, at a flow rate of 1.0 mL/min. Detection was
performed using a photodiode array detector at 258 nm. The method
was validated in accordance with ICH Q2­(R2) guidelines. Validation
parameters, including system suitability, linearity, accuracy, precision,
robustness, sensitivity, and solution stability, were found within
acceptable limits. The proposed method was successfully applied for
the analysis of the stability study of MGF and to analyze the release
kinetics of MGF SLNs, formulated via high-pressure homogenization
and the sonication technique. The results confirm the applicability
of the developed RP-HPLC method for routine quality control of combination
nanocarrier systems.