Qian Meng, Ziqing Li, Tingwei Li, Yun Li, Hua Qu, Wenlong Wei, Qirui Bi, Jianqing Zhang, De-An Guo
The Chrysanthemum genus comprises six predominant varieties (Chuju, Boju, Gongju, Hangju, Huaiju, Jinsihuangju). However, their highly similar morphological and chemical characteristics make accurate discrimination challenging. To address this, a 'Precise-Practical-Rapid' integrated strategy for origin discrimination was developed, incorporating LCHRMS metabolomics (precise), polyphenol-specific HPLC fingerprinting and quantitative analysis (practical), and direct-infusion mass spectrometry and infrared spectroscopy methods (rapid). LC-QTOF MS-based metabolomics identified 221 compounds across six varieties, successfully distinguished all geographical origins, and revealed 13 key discriminatory markers. HPLC-based polyphenol-specific fingerprints and quantitative profiles achieved clear origin clustering, with a minimal four-marker panel (luteolin, rutin, 3-caffeoylquinic acid [3-CQA], and 5-CQA) retaining full discriminatory power. DI-QDa MS and FT‑IR spectroscopy enabled rapid (2 min per sample) high-throughput screening, with t‑SNE, PLS‑DA, and hierarchical cluster analysis achieving successful discrimination. Collectively, these platforms provided complementary resolution tiers: LC-QTOF MS for discovery and traceability in complex matrices, HPLC for routine quality control with reference-standard quantification, and DI-QDa or FT‑IR for on-site screening. This multi-platform comparison establishes a practical decision framework for Chrysanthemum authentication and offers a transferable pipeline for origin verification of other botanicals.