Metabolomic and Lipidomic Profiling of Traditional Chinese Medicine Testudinis Carapax et Plastrum and Its Substitutes

Mengru Xin ¹ Yaodong Ping ² Yisheng Zhang ³ Zhang Wenqing ⁴ Lin Zhang ⁵ Yonghong Zhang ^6,7 Wentao Sheng ⁸ Lei Wang ⁹ Weidong Mao ¹⁰ Ling Xiao ¹¹ Shan Guo ¹² Hankun Hu ^1*

• ¹ Zhongnan Hospital, Wuhan University, Wuhan, China
• ² Key Laboratory of Carcinogenesis and Translational Research, Beijing Cancer Hospital, Peking University, Beijing, Beijing Municipality, China
• ³ Department of Pharmacy, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, China
• ⁴ Department of Pharmacy, Zhongnan Hospital, Wuhan University, Wuhan, Hubei Province, China
• ⁵ Hubei Shizhen Laboratory, Hubei University of Chinese Medicine, Wuhan, Hebei Province, China
• ⁶ Hubei Provincial Key Laboratory of Embryonic Stem Cell Research, Shiyan, China
• ⁷ Laboratory of Medicinal Plant of Hubei Key Laboratory of Embryonic Stem Cell Research, Wuhan, China
• ⁸ Hubei Shengchang Aquatic Products Co., Wuhan, China
• ⁹ Hubei Laozhongyi Pharmaceutical Co., Ltd, Xiaogan, China
• ¹⁰ Georgia Gwinnett College, Lawrenceville, Virginia, United States
• ¹¹ Engineering Research Center for Drug Quality Control, Wuhan, Hebei Province, China
• ¹² Department of Biological Repositories, Zhongnan Hospital, Wuhan University, Wuhan, Hubei Province, China

Chinemys reevesii (Gray) species–sourced Testudinis Carapax et Plastrum (TCP) is an animal-based traditional Chinese medical material, and its decoction or extract possesses multiple pharmacological effects. However, other species-sourced substitutes are sometimes used in the market, potentially impairing the quality and effectiveness of TCP medications. In this study, liquid 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. All 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.