Selective Extraction of 5-Methoxy-6,7-methylenedioxycoumarin from Pterocaulon balansae for Pharmacological Investigation: A Proof-of-Concept Study

5-Methoxy-6,7-methylenedioxycoumarin, the major coumarin found in Pterocaulon balansae Chodat, Asteraceae, has demonstrated cytotoxic activity against glioma cell lines, a highly aggressive brain tumor with limited therapeutic options. To enable preclinical advances, larger quantities of this compound are needed through efficient and selective extraction methods. This study provides a proof-of-concept comparison of three approaches for extracting coumarins from P. balansae , with emphasis on 5-methoxy-6,7-methylenedioxycoumarin: conventional hexane maceration, leaf surface exudate extraction, and direct sublimation at different temperatures (100–180 °C). The extracts were analyzed by ultra-fast liquid chromatography (UFLC) and nuclear magnetic resonance (NMR). The hexane extract yielded seven coumarins, with 5-methoxy-6,7-methylenedioxycoumarin as the predominant constituent. Leaf surface exudate extraction appeared highly selective for 5-methoxy-6,7-methylenedioxycoumarin, while sublimation afforded the highest selectivity and yield among the techniques tested. Therefore, these preliminary results suggest sublimation as the most promising method for scaling up the production of 5-methoxy-6,7-methylenedioxycoumarin to support further biological evaluation, particularly in glioma research.