Journal Article

Development and Validation of Normal Phase HPLC Method for Determination of Vitamin A (Retinyl Palmitate) in Fortified Rice Kernels (FRKs)

Furquani, Julhasnain Mohammad Jubair, Hasan, Md. Rakibul, Begum, Rokeya, Chowdhury, Md. Abu Faysal

2025

Overview

Fortifying rice with vitamin A represents a crucial step toward addressing nutritional deficiencies in Bangladesh. The increasing interest in food enrichment necessitates straightforward and precise methods for analyzing fortified products. This study introduces a simple and reliable approach based on normal‐phase high‐performance liquid chromatography (HPLC) analysis to quantify retinyl palmitate in fortified rice kernels. The method was validated in accordance with the International Conference on Harmonization guidelines. Analysis was conducted using a DB silica column (250 cm × 4.6 mm; 5 μ m), with a mobile phase consisting of n‐hexane and isopropyl alcohol in a 96:04 v / v ratio and a flow rate of 1.0 mL/min. Detection occurred at a wavelength of 326 nm, with a retention time for vitamin A of approximately 3 min and a total run time of 5 min. The calibration plot exhibited a linear relationship ( R 2 = 1.00) across the concentration range of 0.48–30.80 μ g/mL. The limit of detection (LOD) and limit of quantification (LOQ) were 0.015 and 0.051 μ g/mL for standard and 0.35 and 1.07 μ g/mL for spiked samples, respectively. Accuracy, determined through recovery studies, showed a mean recovery rate of over 90%. The relative standard deviation (RSD%) for intraday and interday precision was found to be less than 2%. Robustness and solution stability results were within acceptable limits. The proposed method, meeting all validation criteria including linearity, accuracy, precision, and robustness, offers a rapid and cost‐effective solution for routine quality control of vitamin A‐fortified rice, with potential to support food fortification monitoring in resource‐limited settings.