The tuberous roots of Ophiopogon japonicus and Liriope spicata are used for the same therapeutic purpose in
traditional Chinese medicine and are collectively referred to as maidong medicine. Interestingly, it was observed
that the price of tuberous roots varies depending on their location on the plant, and fibrous roots are usually
discarded post-harvest. Mislabeling might be of concern due to similarities in morphological features between
the two species. Moreover, paclobutrazol has been observed to be heavily applied during the production, and
therefore might be of health concern. Overall, maidong might suffer from quality inconsistencies while its
metabolomic complexity is influenced by growing region and cultivation practices, botanical species, and plant
parts. To address these challenges, this study employed High-Performance Thin Layer Chromatography (HPTLC)
approach, in which sample preparation and derivatization procedure were optimized to enable to capture more
detailed and comprehensive metabolomic fingerprints. By integrating with rTLC algorithm and Multivariate Data
Analysis (MVDA), an improved quality assessment was achieved. Samples were collected from four production
regions and supplemented with commercial products from markets. The optimized HPTLC analysis recognized
species- and region-specific metabolomic patterns of maidong, uncovering a 4% of mislabelled cases. Moreover,
findings highlight the underexplored therapeutic potential of fibrous roots, and comparable therapeutic efficacy
between different root types. Additionally, complemented by Liquid Chromatography-Mass Spectrometry (LCMS) for paclobutrazol residue evaluation, 24.66% of the commercial maidong samples surpassed maximum
residue limits of paclobutrazol, raising safety concerns. This research represents a significant analytical
advancement, offering a robust, cost-effective, and comprehensive method for maidong quality control, and
paving the way for more strict residue regulation and updates to herbal pharmacopoeias and monographs
