A systematic study of Pseudobulbus Cremastrae seu Pleiones: Characteristics, Origin, chemical composition and toxicology.

Ethnopharmacological Relevance: Pseudobulbus Cremastrae seu Pleiones (PCSP) is a multi-source traditional Chinese medicine (TCM) with diverse chemical compositions and toxicity levels. The authenticity identification and safety evaluation of PCSP have attracted widespread attention in clinical applications.

Aim Of This Study: The objective of this study was to evaluate the authenticity and safety of commercially available PCSP.

Materials And Methods: Morphological and microscopic identification, HPLC chromatogram, UPLC-Q-TOF-MS/MS with molecular networking were applied to the authenticity identification of PCSP. The safety of different PCSPs was evaluated by acute toxicity in zebrafish at maximum non-lethal concentration (MNLC) and 10% lethal concentration (LC). Intestinal toxicity of PCSP was assessed through histological staining, intestinal goblet cells, neutrophils, and intestinal opacity.

Results: Four sources of PCSP varied in size, epidermal longitudinal grooves, and microscopic features. GNPS analysis identified 61, 47, 44, and 56 chemical compounds in Cremastra appendiculate (CA), Oreorchis patens (Lindl.) Lindl. (OPL), Iphigenia indica A. Gray (IIG), and Tulipa edulis (Miq.) Baker (TEB). Colchicine and militarine, were discovered as distinguishing markers. Acute toxicity in zebrafish ranked as follows: IIG > OPL > CA > TEB. Further studies on the intestinal toxicity of the authentic PCSP (CA, OPL) showed that CA induced less damage with a smaller lumen area, fewer neutrophils and goblet cells, and reduced peristalsis inhibition compared to OPL, indicating greater safety.

Conclusion: Four different sources of PCSP were accurately distinguished based on three dimensions: character, components, and toxicity. OPL and CA were considered as genuine products, while CA with lower toxicity was more suitable for clinical applications.