Nontargeted metabolomics reveals the discrimination of Cyclocarya paliurus leaves brewed by different methods

An original way of brewing Cyclocarya paliurus leaves, especially in China, is to steep leaves in hot water before people drink it directly. Recently, infusing tea leaves with cold water, a creative way in Taiwan, has become a popular way of making tea. This study was designed to investigate the differences in metabolites among three brewing methods (cold-brewing, steep-brewing, and boil-brewing) based on UPLC-QTOF-MS metabolomics experiments and the feasibility of cold-brewing methods for C. paliurus leaves. Unsupervised analysis (PCA) explained 54.6% (positive ion mode) and 57.4% (negative ion mode) of the total variance, whereas supervised analysis (OPLS-DA) with cross-validated R²Y and Q² values > 0.5, could reveal potential metabolites with better discrimination among the three brewing methods. Fifteen potential differential metabolites were chosen and identified, and nine of them were further confirmed with reference standards. This study suggested that the cold-brewing method without an increase in temperature protected the phenol aromatic ring, thereby obtaining more phenolic acid compounds from C. paliurus leaves. These results provided a basis for making cold tea and promoting the development of cold tea with C. paliurus leaves as raw materials.