Chromatographic Fractionation Protocol of Natural Extracts Using Diaion HP20-SS Resin and Isolera-Biotage System v1

This method processes natural extracts to separate secondary metabolites into distinct chemical classes, supporting their identification and biochemical assessment. A solvent gradient with increasing polarity enables selective elution, improving resolution and compound recovery. A non-ionic polymeric resin was selected for its strong adsorption capacity and versatility in separating compounds of varying polarities. Using the automated Isolera-Biotage system increases reproducibility and efficiency, while reducing experimental time and minimizing material loss. The fractions obtained, being less complex than the crude extract, can be further analyzed by HPLC-DAD, LC-HRMS, and bioactivity assays to provide a broad chemical and functional profile. The approach is applicable to metabolomics, bioprospecting, and bioactive compound isolation, contributing to projects that explore and add value to natural resources, and supporting the discovery of new products with economic, medicinal, or ecological importance.