DEVELOPMENT AND VALIDATION OF ANHPLCMETHODFOR THE QUANTIFICATION OF KEY POLYPHENOLSINRED RICE (Oryza Sativa L.) EXTRACT
Herbal red rice (Oryza sativa L.) is a functional food rich in polyphenols, flavonoids, and anthocyanins that contribute to its antioxidant, anti-inflammatory, and therapeutic properties. Among these, gallic acid, catechin, quercetin, and apigenin are pharmacologically important bioactive compounds. Accurate quantification of thesemarkers is essential for quality control and standardization of red rice-based herbal and nutraceutical formulations. The current study focused on developing and validating a simple, precise, and reproducible high-performance liquidchromatography (HPLC) method for the simultaneous identification and quantification of gallic acid, catechin, quercetin, and apigenin in red rice extract. The extract was prepared using an optimized solvent extraction processwith orbital shaking for 2 hours, followed by spray drying to obtain a powdered sample. Chromatographicseparation was achieved using a reverse-phase C18 column with a gradient mobile phase consisting of ortho- phosphoric acid in water and acetonitrile. Detection was performed at 272 nm, and the retention times of eachanalyte were recorded. The developed method was validated for linearity, sensitivity, precision, robustness, andaccuracy through recovery experiments. Baseline separation was achieved with retention times of 3.2 minutes for gallic acid, 3.5 minutes for catechin, 8.8 minutes for quercetin, and 13.1 minutes for apigenin. The methoddemonstrated excellent linearity, high sensitivity, and good precision, with recovery rates ranging from99 to 102%. Compared with previously reported approaches, this method is faster, more accurate, and specifically optimizedfor standardized herbal and nutraceutical red rice products, thereby offering clear advantages for polyphenol analysisand quality control.
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