Data Sheet 1_Metabolomic and lipidomic profiling of traditional Chinese medicine Testudinis Carapax et Plastrum and its substitutes.docx

BackgroundTestudinis Carapax et Plastrum (TCP), derived from the Chinemys reevesii (Gray) species, is a traditional Chinese medicinal material with demonstrated pharmacological activity in decoctions and extracts. However, other species-sourced substitutes are sometimes used in the market, potentially impairing the quality and effectiveness of TCP medications. To address this issue, systematic characterization of metabolite and lipid profiles in TCP decoctions and their substitutes is critical for establishing quality assessment criteria.
MethodsLiquid chromatography‐tandem mass spectrometry‐based metabolomic and lipidomic analyses were performed to comprehensively characterize metabolites and lipids in water decoctions of genuine TCP and its substitutes, such as Trachemys scripta elegans (Wied)– and Ocadia sinensis (Gray)‐sourced tortoise shells. Absolute quantification of amino acids, derivatives, and small peptides was performed. Statistical analyses were applied to identify significant differences in metabolite and lipid composition across samples.
ResultsAll told, 1117 water-soluble metabolites (including amino acids, organic acids, nucleotides and their metabolites or derivatives, etc.) and 574 organic-soluble lipids (including glycerolipids, sphingolipids, glycerophospholipids, fatty acids, and sterol lipids) were detected in decoctions of TCP and two substitutes. Comparative analyses revealed that there were significantly differential metabolites and lipids among decoctions from different origins, as well as between decoctions of TCP samples and the two substitutes. Of particular interest, the content of N-methyl‐4–aminobutyric acid was lower in the substituted samples than TCP samples. Furthermore, the content of 27 amino acids, 22 amino acid derivatives, and 18 small peptides in the decoctions of TCP and two substitutes were absolutely quantified, constituting up to tens of milligrams per 10 g of tortoise shell. Taken together, our study provides comprehensive metabolomic and lipidomic information for assessing TCP decoction quality.