Urinary metabolomic analysis of three fundamental traditional Mongolian Medicine Constitution types using UHPLC-QTOF-MS and nuclear magnetic resonance techniques

Human constitution is closely linked to health and disease and serves as a basis for disease diagnosis. In this work, the urinary metabolomics of three basic traditional Mongolian medicine constitution (MMC) types were analyzed using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) and nuclear magnetic resonance (NMR) techniques. 90 volunteers majoring in Mongolian medicine from a university during 2021.1–2023.1 were averagely grouped based on the characteristics of MMC using the Classification and Scoring Scale for Mongolian Medicine Constitution: Khii group, Shar group, and Badgan group. NMR technology was employed to determine the urinary metabolites of subjects in different groups. UHPLC-QTOF-MS technology was applied to identify differential metabolites in urine. Multivariate statistical analysis (MSA) including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were adopted to analyze the impacts of urinary metabolites. the positive ion mode exhibited higher signal intensity with better separation efficiency. Accuracy of UHPLC-QTOF-MS and NMR in Mongolian urinary metabolites exceeded 99%. PCA demonstrated satisfactory clustering analysis results. Covered metabolic pathways encompassed tricarboxylic acid cycle (TCA), fatty acid beta-oxidation, as well as protein, nucleic acid, and amino acid metabolisms. the three basic Mongolian constitutions may involve several small molecular metabolites (SMMs) like bile acids, carnitine, aromatic acids, amino acids, and small molecular carboxylic acids. When clinically diagnosing and treating diseases based on Mongolian constitution, constitutional factors should be considered.