Development and Validation of UV-Visible Spectrophotometric and RP-HPLC Methods for Quantification of Mangiferin in Bulk Drug According to ICH Q2(R1) Guidelines

Mangiferin is a bioactive xanthone C-glycoside found highly in Mangifera indica and several medicinal plants. It exhibits promising antioxidant, antidiabetic, anti-inflammatory, and neuroprotective activities. There are only few studies reported the method development of simple, low-cost UV-visible spectrophotometric and RP-HPLC methods with validation parameters suitable for routine quality control of mangiferin in bulk and herbal formulations, revealing a critical research gap in accessible analytical methodologies even though it has reputed pharmaceutical importance. The present study aims to develop and validate rapid, economical and reliable UV-visible spectrophotometric and RP-HPLC methods for the accurate quantification of mangiferin in accordance with ICH Q2(R1) guidelines. In this current research, UV-visible spectrophotometry was performed using methanol and water. RP-HPLC was performed using Shimadzu C8 column (250 mm × 4.6 mm, 5 µm) which was maintained at 25 °C with isocratic elution using 10 mM ammonium acetate buffer (pH 3): acetonitrile (80:20 v/v) as a mobile phase with flow rate of 1 ml/min and detection was performed at 257 nm. Both methods show excellent linearity from 1 to 5 µg/ml with correlation coefficients of 0.999 for UV and 0.9989 for RP-HPLC. The retention time for mangiferin in RP-HPLC method was obtained at 2.9 min. Limits of detection (LOD) and quantification (LOQ) were found to be 5.21 µg/ml and 15.81 µg/ml for UV, and 5.22 µg/ml and 15.81 µg/ml for RP-HPLC, respectively. Precision parameters show %RSD less than 2% for repeatability, intra-day, and inter-day measurements confirming its high reproducibility. All other validation parameters such as ruggedness, robustness, and specificity complied with ICH requirements. As there is an increasing demand for standardized herbal products, these simple, cost-effective and fully validated UV and RP-HPLC methods provide practical approaches for routine quality assurance, batch to batch consistency and regulatory compliance.