Systematic Comparison and Distribution Patterns of Bioactive Metabolites in Different Polarity Fractions of Gentiana scabra Bunge Ethanol‐Water Extract

Fractionation is key for enriching bioactive compounds in traditional Chinese medicine, yet systematic studies on chemical differences across polarity-based fractions are limited. This study systematically compared the chemical profiles of four polarity fractions from a Gentiana scabra Bunge ethanol-water extract to elucidate the distribution patterns of its major active constituents. An integrated approach of untargeted LC-MS and quantitative HPLC was employed. The contents of gentiopicroside, isoorientin, and gentisin generally increased with ethanol concentration, with gentisin showing a more favorable elution trend. Notably, isoorientin and gentisin exhibited co-enrichment specifically in the 50% ethanol fraction (GSE50), resulting in its most complex and balanced chemical profile. This work provides the first systematic elucidation of these distribution patterns, establishing a foundation for precision preparation and quality control based on distinct chemical profiles.