Validación de un método cromatográfico para la determinación cuantitativa polifenoles y flavonoides en extractos vegetales

Abstract

CONDITION FOR PUBLICATION OF PROJECT. This study aims to validate an analytical method for the quantitative determination of polyphenols and flavonoids in plant extracts using Ultra High Performance Liquid chromatography (UPLC) with a diode array detector (DAD). This research is due Ecuador's need to characterize natural bioactive compounds due to its vast biodiversity. The analytical method was developed and validated for the simultaneous separation and quantification of nine target analytes: gallic acid, chlorogenic acid, caffeic acid, syringic acid, p-coumaric acid, rutin, quercetin, naringenin, and kaempferol. The method validation included the assessment of specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) according to the requirements of the ICH standard. The results demonstrated that the method is linear within the working range of 0.5 mg/L–3.0 mg/L, selective, and reproducible, with correlation coefficients above 0.99 in calibration curves and adequate chromatographic resolution for the separation of the analyzed compounds. A recovery rate between 90% and 110% was observed, confirming its precision and applicability in the quantification of polyphenols in plant matrices. This study contributes to the development of reliable techniques for evaluating bioactive compounds in natural matrices. Keywords: Validation, polyphenols, flavonoids, UPLC.