DEVELOPMENT AND VALIDATION OF AN HPLC METHOD FOR QUANTITATIVE DETERMINATION OF MANGIFERIN IN INSTANT TEA CONTAINING DRIED MANGO LEAF EXTRACT
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Abstract
Background: Instant tea containing dried mango leaf extract is a promising product for health-support applications. Mangiferin, a characteristic bioactive compound present in mango leaves, was selected as a chemical marker for product quality control and standardization. The establishment of a fully validated quantitative analytical procedure for mangiferin is therefore a scientific necessity. Objectives: To develop and validate a quantitative method for the determination of mangiferin in instant tea containing dried mango leaf extract using a Shimadzu Prominence-i LC2030C 3D Plus high-performance liquid chromatography system coupled with a photodiode array detector (PDA), in accordance with AOAC and ICH Q2(R2) guidelines. Materials and methods: simulated instant tea samples, mango leaf extract samples, placebo samples. The analytical procedure was developed on an HPLC/PDA system under optimized chromatographic conditions. Validation was conducted across five parameters: system suitability, specificity, linearity, accuracy, and precision. Results: The selected chromatographic conditions included an InertSustain C18 column (150 mm × 4.6 mm, 5 µm) with an isocratic mobile phase consisting of acetonitrile and 0.2% phosphoric acid solution (15:85, v/v), a flow rate of 1.0 mL/min, and an injection volume of 10 µL. The method met system suitability requirements and demonstrated good specificity. The linearity range was 19.90– 79.59 µg/mL with a correlation coefficient (r²) greater than 0.999. The recovery ranged from 97.30% to 99.50%, and the precision showed an RSD of 1.1%, meeting validation acceptance criteria. The mangiferin content determined in the instant tea test sample reached 99.50% of the theoretical value (RSD = 1.1%, n = 18). Conclusion: The developed method for mangiferin quantification demonstrated adequate reliability and complies with validation requirements. The method is suitable for routine quality control testing and standardization of instant tea products containing dried mango leaf extract.
Keywords
Mangiferin, HPLC/PDA, dried mango leaf extract, instant tea
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