Unlocking the chemical blueprint of Gastrodiae Rhizoma variants and hybrids: A pseudotargeted LC-MS/MS metabolomics approach for cultivar authentication

Gastrodiae Rhizoma (Tianma, GR), the dried tuber of Gastrodia elata Bl., is a valuable herb for medicinal and dietary applications. Current markets feature three key cultivars: G. elata f. glauca (WTM), G. elata f. elata (HTM), and their hybrid (WHTM), with significant price variations yet lacking defined chemical authentication markers. Here, we implemented a pseudotargeted LC-MS/MS metabolomics strategy employing 534 optimized MRM transitions. This strategy was coupled with multivariate analysis – unsupervised principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) – of 40 authenticated batches, to decipher cultivar-specific chemical fingerprints. The approach identified 88 statistically validated markers, with 14 pivotal discriminators—primarily 4-Hydroxybenzyl alcohol derivatives—exhibiting distinct cultivar distribution patterns. This chemical blueprint serves as a tool for authentication in quality control and supports regulatory standard establishment for GR products.