UPLC-ESI-MS/MS analysis revealed the dynamic changes and conversion mechanism of non-volatile metabolites during green tea fixation

Fixation is crucial for green tea quality; previous studies were conducted on finished teas and focused on the volatile metabolites of green tea. However, non-volatile metabolites' (NVMs) evolution and formation mechanisms during fixation remain unclear. Here, fixated tea samples were collected at seven different time points during the green tea fixation stage, the dynamic changes and formation mechanisms of NVMs investigated via ultra-performance liquid chromatography-tandem mass spectrometry and enzymatic activity analyses. The results showed that fixation contained enzymatic activity thermal and non-enzymatic thermal stages. Moreover, 1196 NVMs (10 classes) were detected, and significant NVM variations were observed during the "enzymatic activity thermal" fixation stages, with increasing trends in lipids and amino acids, except alkaloids. Furthermore, 160 common differential NVMs were identified. KEGG analysis revealed that unsaturated fatty acid, linoleic acid phenylpropanoid, and flavonoid metabolism pathways were essential during fixation and influenced green tea aroma, taste, and color formation. These results provide a theoretical basis to regulate green tea fixation technology and enrich the chemical theory of green tea processing.