Structure-guided design of adsorbent: Online SPE-semi-preparative HPLC for efficient isolation and purification of three praeruptorins from Peucedanum Praeruptorum Dunn.

A novel ester-functionalized porous polymer adsorbent with a specific surface area of 268.5 m2/g was rationally synthesized via a one-step strategy, which enables selective adsorption of target coumarins relying on multiple intermolecular interactions (hydrogen bonding, hydrophobic effect, π-π stacking, etc.). An integrated online solid-phase extraction-semi-preparative HPLC system was established by coupling this tailor-made adsorbent with semi-preparative HPLC, realizing automated one-step extraction, enrichment and purification of target compounds directly from crude extracts. The loading volume and washing procedures were systematically investigated, and the optimal conditions were determined as follows: a sample loading volume of 3 mL, followed by washing with 0.1% aqueous phosphoric acid at a flow rate of 4.7 mL/min for 2 min. The integrated system exhibited excellent separation performance: praeruptorin A was obtained with 98% purity and a yield of 6.69 mg/g herb, and satisfactory isolation efficiency was achieved for praeruptorin B and praeruptorin C. Impressively, the adsorbent remained stable adsorption performance after 100 reuse cycles, significantly improving the cost-effectiveness of the purification process. This work provides a highly efficient and selective strategy for purifying bioactive coumarins from Peucedanum praeruptorum Dunn, and offers a versatile methodological blueprint and functional adsorbent platform for targeted isolation of high-value components from complex natural matrices, which will promote the research and development of natural product-based drug discovery and herbal medicine.