Comprehensive Saponin Profiling and Comparison of Ten Panax Species Using a Universal Offline Two-Dimensional LC–MS Platform

Panax species are prized globally for tonic effects and widely used in health foods and herbal products, yet genus-wide comprehensive chemical profiling remains limited. Here, we present the first comparative chemical analysis encompassing nearly all Panax species distributed in China, totaling ten species. To achieve this, an offline two-dimensional liquid chromatography–mass spectrometry approach applicable across ten species was established, yielding orthogonality A0 values of 0.65–0.78. Using UNIFI-based automated annotation and manual validation, 4080 compounds (397–1016 compounds per species) were identified─representing the largest saponin data set to date. Comparative analysis across ten species revealed distinct chemotypic patterns: five species were dominated by PPD-type, two by PPT-type, and three by OA-type saponins. Saponins bearing two or three glycosyl groups predominated, with ∼20% showing acylation. P. vietnamensis exhibited the most diverse glycosylation and acylation patterns. This study provides a valuable chemical resource for advancing Panax research and authentication.