Identification of putative biomarker molecules within the genus Corydalis (Papaveraceae) was pursued by combining conventional off-line sample enrichment with high-performance liquid chromatography–solid phase extraction–nuclear magnetic resonance (HPLC–SPE–NMR) based structure elucidation. Off-line reversed phase solid phase extraction (SPE) was used to enrich the desired analytes from a methanolic extract (93mg dry weight) of a miniscule single tuber (233mg dry weight) of C. solida. An aliquot of the SPE fraction (2.1mg) was subjected to separation in the HPLC–SPE–NMR hyphenation. Chromatographic peaks bearing the metabolites under investigation were trapped in the SPE device in a single experiment and transferred to a 600MHz NMR spectrometer equipped with a 30μl cryofit insert fed into a 3mm cryoprobe. Recorded homo- and heteronuclear 1D and 2D NMR data allowed the identification of the three analytes under investigation as protopine, allocryptopine, and N-methyl-laudanidinium acetate. The latter is a rare alkaloid, which has been isolated only once before.