Metabolomics analyses of traditional Chinese medicine formula Shuang Huang Lian by UHPLC-QTOF-MS/MS

BACKGROUND: Shuang Huang Lian (SHL) is a traditional Chinese medicine (TCM) formula made from Lonicerae Japonicae Flos, Forsythiae Fructus, and Scutellariae Radix. Despite the widespread use of SHL in clinical practice for treating upper respiratory tract infections (URTIs), the complete component fingerprint and the pharmacologically active components in the SHL formula remain unclear. The objective of this study was to develop an untargeted metabolomics method for component identification, quantitation, pattern recognition, and cross-comparison of various SHL preparation forms (i.e., granule, oral liquid, and tablet). METHODS: Ultra-high-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS/MS) together with bioinformatics were used for chemical profiling, identification, and quantitation of SHL. Multivariate data analyses such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were performed to assess the correlations among the three SHL preparation forms and the reproducibility of the technical and biological replicates. RESULTS: A UHPLC-QTOF-MS/MS-based untargeted metabolomics method was developed and applied to analyze three SHL preparation forms, consisting of 178 to 216 molecular features. Among the 95 common molecular features from the three SHL preparation forms, quantitative analysis was performed using a single exogenous reference internal standard. Forty-seven of the 95 common molecular features have been identified using various databases. Among the 47 common components, there were 17 flavonoids, 7 oligopeptides, 5 terpenoids, 2 glycosides, 2 cyclohexanecarboxylic acids, 2 spiro compounds, 2 lipids, 2 glycosylglycerol derivatives, and 8 various compounds such as alkyl caffeate ester, aromatic ketone, benzaldehyde, benzodioxole, benzofuran, chalcone, hydroxycoumarin, and purine nucleoside. Five of the 47 common components were designated by the Chinese Pharmacopoeia as the quality markers of medicinal plants of SHL, and 15 were previously reported to have pharmacological activities. Distinct patterns of the three SHL preparation forms were observed in the PCA and PLS-DA plots. CONCLUSIONS: The developed method is reliable and reproducible, which is useful for the profiling, component identification, quantitation, quality assessment of various SHL preparation forms and may apply to the analysis of other TCM formulas. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13020-022-00610-x.