Enhancing and accelerating natural product profiling: The power of CPC, HPLC, HRMS/MS, and NMR synergy

The inherent chemical complexity of natural extracts poses a significant challenge to the rapid and confident chemical profiling and annotation of bioactive compounds [ 1 ]. To tackle this issue, we have developed integrated workflows that leverage the strengths of Centrifugal Partition Chromatography (CPC), High-Performance Liquid Chromatography (HPLC), High-Resolution Tandem Mass Spectrometry (HRMS/MS), and Nuclear Magnetic Resonance (NMR). Together, these methodologies create a synergy that enhances the speed, accuracy, and depth of natural product profiling [ 2 ] [ 3 ]. This keynote will delve into how CPC, as a support-free fractionation technique, is the technique of choice for the chemical deconvolution of natural product mixture while preserving labile compounds, thereby establishing optimal conditions for subsequent spectroscopic analyses. When paired with HPLC and online bioassays, such as DPPH-based antioxidant screening, CPC significantly expedites bioactivity-guided fractionation. HRMS/MS and 13C NMR are then employed in conjunction for dereplication, supported by in silico-predicted and curated databases (VersaDB) [ 4 ]. These datasets are integrated and visualized using tools like Molecular Networking and heat maps. The various annotations are then classified according to their degree of confidence using the CATHEDRAL tool. Through case studies, we will illustrate the effectiveness of this multimodal strategy in rapidly annotating both major and minor constituents. Special emphasis will be placed on the complementary relationship between MS and NMR data, which enhances structural annotation and fosters the discovery of new bioactive natural products. Publication History Article published online: 05 March 2026 © 2026. Thieme. All rights reserved. Georg Thieme Verlag KG Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany