Exploring the Differences in Alkaloids of Fritillariae Thunbergii Bulbus From Different Regions Using Liquid Chromatography‐Mass Spectrometry and Chemometrics

The geographical origin of Fritillariae Thunbergii Bulbus (FTB) is a critical determinant of its quality and efficacy, representing a long-standing challenge in quality control and market standardization. This persistent issue necessitates the development of reliable analytical methods for accurate origin authentication. To address this, we developed an integrated analytical strategy utilizing liquid chromatography coupled with mass spectrometry (LC-MS). This strategy synergistically combines targeted and non-targeted analytical approaches with multivariate statistical analysis to accurately differentiate between FTB produced in Zhejiang province (FPZ) and that from other regions (non-Zhejiang-produced FTB, FNZ). Moreover, a key innovation of this study is the integration of modern chemical profiling with traditional morphological assessment in a synergistic manner. This integrated approach establishes a direct correlation between externally observable traits of the plant and its intrinsic quality parameters, providing a more comprehensive and scientifically grounded framework for quality evaluation. This comprehensive strategy concurrently evaluates both external morphological characteristics and internal chemical profiles, and is executed through a dual-platform analytical workflow. First, ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS) was used to precisely quantify three key alkaloids: verticine, verticinone, and peimisine. Concurrently, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS), integrated with chemometric analysis, was utilized for non-targeted alkaloid profiling to identify characteristic components distinguishing FPZ from FNZ. Structural characterization and semi-quantitative analysis of the differential components identified 11 alkaloids as characteristic chemical markers. Notably, the relative levels of these markers showed significant correlations with key external morphological features. Collectively, these findings provide a scientific foundation for optimizing the standardized cultivation of FTB and for ensuring the consistency of its clinical efficacy.